Methods for identifying risk of melanoma and treatments thereof

Abstract
Provided herein are methods for identifying risk of melanoma in a subject and/or subjects at risk of melanoma, reagents and kits for carrying out the methods, methods for identifying candidate therapeutics for treating melanoma, therapeutic methods for treating melanoma in a subject and compositions comprising one or more melanoma cells and one or more NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed agents. These embodiments are based upon an analysis of polymorphic variations in a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, exemplified by nucleotide sequences of SEQ ID NO: 1-17.
Description
FIELD OF THE INVENTION

The invention relates to genetic methods for identifying risk of melanoma and treatments that specifically target the disease.


BACKGROUND

In some parts of the world, especially among western countries, the number of people who develop melanoma is increasing faster than any other cancer. In the United States, for example, the number of new cases of melanoma has more than doubled in the past twenty years. The probability of developing melanoma increases with age, but this disease effects people of all age groups. Melanoma is one of the most common cancers in young adults.


Melanoma occurs when melanocytes (pigment cells) become malignant. Most pigment cells are in skin, and when melanoma begins its etiology in the skin it is referred to as coetaneous melanoma. Melanoma may also occur in the eye and is called ocular melanoma or intraocular melanoma. Rarely, melanoma arises in the meninges, the digestive tract, lymph nodes or other areas where melanocytes are found. Within the skin, melanocytes are located throughout the lower part of the epidermis, the latter being the surface layer of the skin. Melanocytes produce melanin, which is the pigment that gives skin its natural color. When skin is exposed to the sun, melanocytes produce more pigment, causing the skin to tan or darken.


Sometimes, clusters of melanocytes and surrounding tissue form benign growths referred to as moles or nevi (singular form is nevus). Cells in or near the nevi can divide without control or order and form malignant tumors. When melanoma spreads, cancer cells often are found in the lymph nodes. If the cancer has reached the lymph nodes, it may mean that cancer cells have spread to other parts of the body such as the liver, lungs or brain, giving rise to metastatic melanoma.


Melanoma is currently diagnosed by assessing risk factors and by performing biopsies. Risk factors for melanoma are a family history of melanoma, the presence of dysplastic nevi, patient history of melanoma, weakened immune system, many ordinary nevi, exposure levels to ultraviolet radiation, exposure to severe sunburns especially as a child or teenager, and fair skin. In a biopsy, a pathologist typically examines the biopsied tissue under a microscope to identify cancer cells. Depending upon the thickness of a tumor, if one exists, a physician may order chest x-ray, blood tests, liver scans, bone scans, and brain scans to determine whether the cancer spread to other tissues. Also, a test that identifies p16 nucleotide sequences is sold.


Upon a diagnosis of melanoma, the standard treatment is surgery. Side effects of surgery typically are pain and scarring. Surgery is generally not effective, however, in controlling melanoma that is known to have spread to other parts of the body. In such cases, physicians may utilize other methods of treatment, such as chemotherapy, biological therapy, radiation therapy, or a combination of these methods. Chemotherapy agents for treating melanoma include cisplatin, vinblastine, and dacarbazine. Chemotherapy can lead to side effects such as an increased probability of infection, bruising and bleeding, weakness and fatigue, hair loss, poor appetite, nausea and vomiting, and mouth and lip sores. Side effects of radiation therapy include fatigue and hair loss in the treated area. Biological therapies currently utilized for treatment of melanoma include interferon and interleuken-2. Side effects caused by biological therapies include flu-like symptoms, such as chills, fever, muscle aches, weakness, loss of appetite, nausea, vomiting, and diarrhea; bleeding and bruising skin; rashes, and swelling.


Certain melanoma therapeutics are in clinical trials. For example, canvaxin, which is a whole cell allogenic vaccine developed by irradiating tumor cells from two different patients, is under study. In addition, MAGE-1 and 3 minigenes and peptides and gp 100 peptides are being tested. Upcoming studies include testing of agents such as dacarbazine with a bcl-2 antisense oligonucleotide, and paclitaxel in combination with a matrix metalloprotease inhibitor.


SUMMARY

It has been discovered that polymorphic variations in NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO and/or REPS2 loci in human genomic DNA are associated with occurrence of melanoma. Thus, featured herein are methods for identifying a subject at risk of melanoma and/or determining risk of melanoma in a subject, which comprise detecting the presence or absence of one or more polymorphic variations associated with melanoma in a nucleic acid sample from the subject. The one or more polymorphic variations often are detected in or near a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence, which are set forth as SEQ ID NO: 1-17 respectively, or a substantially identical nucleotide sequence thereof.


Also featured are nucleic acids that encode a NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, and include one or more polymorphic variations associated with melanoma, and oligonucleotides which hybridize to those nucleic acids. Also provided are polypeptides encoded by nucleic acids having a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence, which include the full-length polypeptide, isoforms and fragments thereof. In an embodiment, an NID2 protein is encoded which includes a serine at amino acid position 656. In addition, featured are methods for genotyping a nucleic acid from an individual at positions described herein, methods for identifying candidate therapeutic molecules for treating melanoma and related disorders, as well as methods of treating melanoma in a subject by administering a therapeutic molecule.


Also provided are compositions comprising a melanoma cell and/or a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, or a fragment or substantially identical nucleic acid thereof, with a RNAi, siRNA, a complementary or antisense DNA or RNA, or ribozyme nucleic acid designed from a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence. In an embodiment, the nucleic acid is designed from a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence that includes one or more melanoma associated polymorphic variations, and in some instances, specifically interacts with such a nucleotide sequence. Further, provided are arrays of nucleic acids bound to a solid surface, in which one or more nucleic acid molecules of the array are NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids, or a fragment or substantially identical nucleic acid thereof, or a complementary nucleic acid of the foregoing. Featured also are compositions comprising a melanoma cell and/or a protein, polypeptide or peptide encoded by a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid with an antibody that specifically binds to the protein, polypeptide or peptide. In an embodiment, the antibody specifically binds to an epitope in a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 protein, polypeptide or peptide that includes a non-synonymous amino acid modification associated with melanoma, such as a serine at position 656 of a NID2 protein, polypeptide or peptide.




BRIEF DESCRIPTION OF THE DRAWINGS


FIGS. 1A, 1B and 1C show proximal SNPs in and around a NRP1 region for males and females combined, females alone, and males alone, respectively. FIGS. 2A, 2B and 2C show proximal SNPs in and around a ENDO180 region for males and females combined, females alone, and males alone, respectively. FIGS. 3A, 3B and 3C show proximal SNPs in and around a CDK10 region for males and females combined, females alone, and males alone, respectively. FIGS. 4A, 4B and 4C show proximal SNPs in and around a PCLO region for males and females combined, females alone, and males alone, respectively. FIGS. 5A, 5B and 5C show proximal SNPs in and around a FPGT/CARK region for males and females combined, females alone, and males alone, respectively. FIGS. 6A, 6B and 6C show proximal SNPs in and around a REPS2 region for males and females combined, females alone, and males alone, respectively. The position of each SNP on the chromosome is shown on the x-axis and the y-axis provides the negative logarithm of the p-value comparing the estimated allele to that of the control group. Also shown are exons and introns of the genes in the approximate chromosomal positions.



FIG. 7A depicts effects of NRP1-directed siRNA on melanoma M14 cell line proliferation according to a WST 1 assay. FIG. 7B shows effects of NRP1-directed siRNA on melanoma A375 cell line proliferation according to a WST 1 assay.



FIG. 8 depicts effects of CDK10_siRNA on melanoma A375 cell line proliferation according to a WST-1 assay.




DETAILED DESCRIPTION

It has been discovered that polymorphic variants in and around NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequences are associated with occurrence of melanoma in subjects. Thus, detecting genetic determinants associated with an increased risk of melanoma occurrence can lead to early identification of melanoma or susceptibility to melanoma and early prescription of preventative measures and treatments. Also, associating NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polymorphic variants with melanoma has provided new targets for screening molecules useful in melanoma prognostics/diagnostics and melanoma treatments.


Melanoma and Sample Selection


Melanoma is typically described as a malignant neoplasm derived from cells capable of forming melanin. Melanomas arise most commonly in the skin of any part of the body, or in the eye, and rarely, in the mucous membranes of the genitalia, anus, oral cavity, or other sites. Melanoma occurs mostly in adults and may originate de novo or from a pigmented nevus or lentigo maligna. Melanomas frequently metastasize widely to regions such as lymph-nodes, skin, liver, lungs, and brain.


In the early phases, the cutaneous form is characterized by proliferation of cells at the dermal-epidermal junction that soon invade adjacent tissues. The cells vary in amount and pigmentation of cytoplasm; the nuclei are relatively large and irregular in shape, with prominent acidophilic nucleoli; and mitotic figures tend to be numerous. Other criteria for melanomas are asymmetry, irregular borders, heterogeneous color, large diameter, and a recent change in shape, size or pigmentation. Excised melanoma skin samples are often subjected to the following analyses: diagnosis of the melanocytic nature of the lesion and confirmation of its malignancy; maximum tumor thickness in millimeters (according to Breslow's method); assessment of completeness of excision of invasive and in situ components and microscopic measurements of the shortest extent of clearance; level of invasion (Clark); presence and extent of regression; presence and extent of ulceration; histological type and special variants; pre-existing lesion; mitotic rate; vascular invasion; neurotropism; cell type; tumor lymphocyte infiltration; and growth phase, vertical or radial.


Based in part upon selection criteria set forth above, individuals having melanoma can be selected for genetic studies. Also, individuals having no history of cancer or melanoma often are selected for genetic studies. Other selection criteria can include: a tissue or fluid sample is derived from an individual characterized as Caucasian; a sample is derived from an individual of German paternal and maternal descent; and relevant phenotype information is available for the individual. Phenotype information corresponding to each individual can include sex of the individual, number of nevi (e.g., actual number or relative number (e.g., few, moderate, numerous)), hair color (e.g., black, brown, blond, red), diagnosis of melanoma (e.g., tumor thickness, date of primary diagnosis, age of individual as of primary diagnosis, post-operative tumor classification, presence of nodes, occurrence of metastases, subtype, location), country or origin of mother and father, presence of certain conditions for each individual (e.g., coronary heart disease, cardiomyopathy, arteriosclerosis, abnormal blood clotting/thrombosis, emphysema, asthma, diabetes type 1, diabetes type 2, Alzheimer's disease, epilepsy, schizophrenia, manic depression/bipolar disorder, autoimmune disease, thyroid disorder, and hypertension), presence of cancer in the donor individual or blood relative (e.g., melanoma, basaliom/spinaliom/lentigo malignant/mycosis fungoides, breast cancer, colon cancer, rectum cancer, lung cancer, lung cancer, bronchus cancer, prostate cancer, stomach cancer, leukemia, lymphoma, or other cancer in donor, donor parent, donor aunt or uncle, donor offspring or donor grandparent).


Provided herein is a set of blood samples and a set of corresponding nucleic acid samples isolated from the blood samples, where the blood samples are donated from individuals diagnosed with melanoma. The sample set often includes blood samples or nucleic acid samples from 100 or more, 150 or more, or 200 or more individuals having melanoma, and sometimes from 250 or more, 300 or more, 400 or more, or 500 or more individuals. The individuals can have parents from any place of origin, and in an embodiment, the set of samples are extracted from individuals of German paternal and German maternal ancestry. The samples in each set may be selected based upon five or more criteria and/or phenotypes set forth above.


Polymorphic Variants Associated with Melanoma


A genetic analysis described hereafter linked melanoma with polymorphic variants of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 from human subjects. Genomic nucleotide sequences representative of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids are set forth in SEQ ID NOS: 1-7, respectively, and are incorporated herein by reference from published database entries (see Examples section and http address at www.ncbi.nlm.nih.gov/LocusLink/). The following is a description of molecules encoded by such regions.


NRP1


NRP1 is known as neuropilin 1, NRP, VEGF165R (Vascular Endothelial Growth Factor-165 Receptor). NRP1 protein is 923 amino acids in length and the corresponding genomic nucleotide sequence has been mapped to chromosomal position 10p 12.


Neuropilin is a type I transmembrane protein and is an epitope recognized by a monoclonal antibody that labels specific subsets of axons in the developing Xenopus nervous system. Neuropilin comprises in its extracellular domain several distinctive motifs; its cytoplasmic domain is short (40 amino acids) and is highly conserved among Xenopus, mouse, and chick. An aberrant retinal pathway and visual centers in Xenopus tadpoles share a common cell surface molecule, A5 antigen (Dev. Biol. 135: 231-240, 1989). The gene encoding human neuropilin was cloned and the structure of the protein product was characterized (He & Tessier-Lavigne, Neuropilin is a receptor for the axonal chemorepellent semaphorin III. Cell 90: 739-751, 1997).


A VEGF receptor (NCBI MIM 192240) was discovered which reportedly binds VEGF165 but not VEGF121. This isoform-specific VEGF receptor (VEGF165R) is identical to human neuropilin-1, which is also a receptor for the collapsin/semaphorin family that mediates neuronal cell guidance (Soker et al., Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92: 735-745, 1998). It also was shown that when coexpressed in cells with KDR (VEGFR2; NCBI MIM 191306), neuropilin-1 enhances the binding of VEGF165 to KDR and VEGF165-mediated chemotaxis. Conversely, inhibition of VEGF165 binding to neuropilin-1 inhibits its binding to KDR and its mitogenic activity for endothelial cells. It was proposed that neuropilin-1 is a novel VEGF receptor that modulates VEGF binding to KDR and subsequent bioactivity and therefore may regulate VEGF-induced angiogenesis.


It was concluded that NRP1-mediated interactions are a necessary element in the initiation of the primary immune response and offer another example, like that of agrin (NCBI MIM 103320), of a molecule shared by neurologic and immunologic synapses (Tordjman et al., A neuronal receptor, neuropilin-1, is essential for the initiation of the primary immune response. Nature Immun. 3: 477-482, 2002). It was shown that VEGF selectively upregulates NRP1 but not NRP2 via the VEGF receptor 2-dependent pathway. In a murine model of VEGF-dependent angioproliferative retinopathy, intense NRP1 mRNA expression was observed in the newly formed vessels. Furthermore, selective NRP1 inhibition in this model suppressed neovascular formation substantially. These results suggested that VEGF can not only activate endothelial cells directly but also contribute to robust angiogenesis in vivo by a mechanism that involves upregulation of its cognate receptor expression (Oh et al., Selective induction of neuropilin-1 by vascular endothelial growth factor (VEGF): a mechanism contributing to VEGF-induced angiogenesis. Proc. Nat. Acad. Sci. 99: 383-388, 2002).


Transgenic mice died in utero at embryonic day 8.5 when both NRP1 and NRP2, which they called Np1 and Np2, respectively, were knocked out. The yolk sacs of these mice were avascular. Mice deficient for NRP2 but heterozygous for NRP1 or deficient for NRP1 but heterozygous for NRP2 were also embryonic lethal and survived to embryonic days 10 to 10.5. Other details of the abnormal vascular phenotype resembled those of VEGF and Vefgr2 knockouts. The results suggested that neuropilins are early genes in embryonic vessel development and that both NRP1 and NRP2 are required (Takashima et al., Targeting of both mouse neuropilin-1 and neuropilin-2 genes severely impairs developmental yolk sac and embryonic angiogenesis. Proc. Nat. Acad. Sci. 99: 3657-3662, 2002).


It was shown in zebrafish the NRP1 protein was a functional receptor for human VEGF165. Whole-mount in situ hybridization showed that transcripts of the zebrafish NRP1 gene during embryonic and early larval development were detected mainly in neuronal and vascular tissues. A knockdown of the gene in embryos resulted in vascular defects. Embryos treated with VEGFR2 kinase inhibitor had a similar vessel defect, suggesting that knockdown of zebrafish NRP1 reduces VEGF activity. To determine whether NRP1 and VEGF activities are interdependent in vivo, zebrafish NRP1 and VEGF morpholinos were coinjected into embryos at concentrations that individually did not significantly inhibit blood vessel development. The result was a potent inhibition of blood cell circulation via both intersegmental and axial vessels, demonstrating that VEGF and NRP1 act synergistically to promote a functional circulatory system. These results provided the first physiologic demonstration that NRP1. regulated angiogenesis through a VEGF-dependent pathway (Lee et al., Neuropilin-1 is required for vascular development and is a mediator of VEGF-dependent angiogenesis in zebrafish. Proc. Nat. Acad. Sci. 99: 10470-10475, 2002).


NID2


The protein Nidogen 2 is also known as NID2 or osteonidogen. NID2 protein is 1376 amino acids in length and has a predicted molecular weight of 151,153 daltons (but see below). NID2 is a member of the nidogen superfamily and has EGF homology; LDL receptor YWTD-containing repeat homology; and thyroglobulin type I repeat homology. The NID2 genomic sequence has been mapped to chromosomal position 14q21-q22.


Basement membranes, which are composed of type IV collagens (see NCBI MIM 120130), laminins (see LAMC1; NCBI MIM 150290), perlecan (HSPG2; NCBI MIM 142461), and nidogen (see NID1; NCBI MIM 131390), are thin pericellular protein matrices that control a large number of cellular activities, including adhesion, migration, differentiation, gene expression, and apoptosis. By sequencing several overlapping cDNA clones in both directions, a cDNA encoding NID2 was obtained Kohfeldt et al., Nidogen-2: a new basement membrane protein with diverse binding properties. J. Molec. Biol. 282: 99-109, 1998). Sequence analysis predicted that the 1,375-amino acid NID2 protein, which is 46% identical to NID1, contains a 30-residue signal peptide, 49 primarily central cys residues, 5 potential N-linked glycosylation sites, 2 tyr residues in a potential O-sulfation sequence, and a YGD rather than an RGD cell adhesion sequence. Electron microscopy confirmed the presence of 3 deduced globular domains connected by a link and a rod-like region. Additional predicted structures similar to those found in NID1 include 5 epidermal growth factor (EGF; NCBI MIM 131530)-like modules, 2 of which have potential calcium-binding sequences; 2 thyroglobulin (NCBI MIM 188450) type I modules; and 5 low density lipoprotein (LDL) receptor (NCBI MIM 606945) modules. SDS-PAGE analysis showed that NID2 is expressed as a 200-kD protein, larger than the calculated mass of 148 kD, presumably due to oligosaccharide substitution as indicated by hexosamine analysis. Northern blot analysis revealed ubiquitous expression of a 5.5-kb NID2 transcript that was strongest in heart and placenta, moderate in pancreas, kidney, and skeletal muscle, and weakest in brain. Immunoblot analysis detected expression of NID2 in muscle, heart, placenta, kidney, skin, and testis, with weaker expression in liver and brain. Immunofluorescence analysis of mouse tissues showed staining of basement membranes usually in close colocalization with NID 1.


It was determined that the mouse NID2 gene contains 21 exons (Schymeinsky et al., Gene structure and functional analysis of the mouse nidogen-2 gene: nidogen-2 is not essential for basement membrane formation in mice. Molec. Cell. Biol. 22: 6820-6830, 2002). The promoter region contains several putative SP1 (NCBI MIM 189906) and CAAT recognition sites, but lacks a TATA box. Mice carrying a phenotypic null mutation in the NID2 gene have been developed. NID2-deficient mice showed no overt abnormalities, and their basement membranes appeared normal by ultrastructural analysis and immunostaining. NID2 deficiency did not lead to hemorrhages, and NID2 did not appear essential for basement membrane formation or maintenance.


ENDO180


The ENDO180 nucleotide sequence encodes a protein also known and sometimes referenced herein as UPARAP (urokinase plasminogen activator receptor-associated protein), KIAA0709, MRC2 (mannose receptor, C type 2), endocytic receptor (macrophage mannose receptor family) and is described as a novel mesenchymally expressed member of the macrophage mannose receptor family of endocytic receptors. ENDO180 is 1479 amino acids in length and has a molecular weight of about 180 kDa. ENDO180 has been mapped to chromosomal position 17q24.1.


ENDO180 is a member of the mannose receptor family (East et al., Biochim Biophys Acta. 2002 Sep. 19; 1572(2-3): 364-86. The mannose receptor family comprises four glycoproteins each of which is a type I transmembrane receptor with an N-terminal cysteine-rich domain, a single fibronectin type II (FNII) domain and eight to ten C-type lectin-like domains (CTLDs). Characteristically, these proteins are able to recycle between the plasma membrane and the endosomal apparatus due to discrete motifs present within their cytoplasmic domains. The family includes mannose receptor (MR), M-type receptor for secretory phospholipases A(2) (PLA(2)R), DEC-205/gp200-MR6 and ENDO180/uPARAP. Despite their overall structural similarity, these four receptors have evolved to use different domains to interact with discrete ligands. In addition, they differ in their ability to mediate endocytic and phagocytic events and in their intracellular destinations. Together, they represent a unique group of multidomain, multifunctional receptors.


Members of this receptor family are unusual in that they contain 8-10 C-type lectin carbohydrate recognition domains in a single polypeptide backbone, however, only the macrophage mannose receptor had been shown to function as a lectin (Sheikh et al., J Cell Sci. 2000 March; 113 (Pt 6): 1021-32). Sequence analysis of ENDO180 reveals that the second carbohydrate recognition domain has retained key conserved amino acids found in other functional C-type lectins. The protein also displays Ca(2+)-dependent binding to N-acetylglucosamine but not mannose affinity columns. ENDO180 predominantly is expressed in vivo and in vitro on fibroblasts, endothelial cells and macrophages, and the distribution and post-translational processing in these cells is consistent with ENDO180 functioning to internalize glycosylated ligands from the extracellular milieu for release in an endosomal compartment.


The uptake and lysosomal degradation of collagen by fibroblasts constitute a major pathway in the turnover of connective tissue (Engelholm, et al., J. Cell Biol. 2003 Mar. 31; 160(7): 1009-15). It was reported that ENDO180 is a key player in this process. Fibroblasts from mice with a targeted deletion in the ENDO180 gene displayed a near to complete abrogation of collagen endocytosis. These cells had diminished initial adhesion to a range of different collagens, as well as impaired migration on fibrillar collagen.


CDK10


The protein CDK10 (cyclin-dependent kinase (CDC2-like) 10) is also known as cyclin-dependent kinase 10 isoform 1 (331 amino acids); cyclin-dependent kinase 10 isoform 2 (314 amino acids); cyclin-dependent kinase 10 isoform 3 (123 amino acids); CDC2-related protein kinase; cell division protein kinase 10; cyclin-dependent kinase related protein; serine/threonine protein kinase and PISSLRE. CDK10 has been mapped to chromosomal position 16q24.


The protein encoded by CDK10 belongs to the CDK subfamily of the Ser/Thr protein kinase family. The CDK subfamily members are highly similar to the gene products of S. cerevisiae cdc28, and S. pombe cdc2, and are known to be essential for cell cycle progression. This kinase has been shown to play a role in cellular proliferation. Its function is limited to cell cycle G2-M phase. At least three alternatively spliced transcript variants encoding different isoforms have been reported, two of which contain multiple non-AUG translation initiation sites. Cyclin-dependent kinases (CDKs) are CDC2 (NCBI MIM 116940)-related kinases that bind to cyclin to form active holoenzymes that play a pivotal role in the regulation of the eukaryotic cell cycle. A 360-amino acid protein PISSLRE was predicted based on the amino acid sequence of the region corresponding to the conserved CDC2 PSTAIRE motif. PISSLRE contains all the structural elements characteristic of CDKs and unique extensions at both ends. Sequence comparisons revealed that it shares 41% and 50% protein sequence identity with CDC2 and CDC2L1 (NCBI MIM 176873), respectively. It was determined that PISSLRE was expressed broadly in human tissues as a 2-kb mRNA. An additional 3.5-kb transcript was observed in some tissues (Brambilla et al., Molecular cloning of PISSLRE, a novel putative member of the cdk family of protein serine/threonine kinases. Oncogene 9: 3037-3041, 1994).


FPGT


The protein FPGT (fucose-1-phosphate guanylyltransferase) is also known as GFPP and GDP-beta-L-fucose pyrophosphorylase. FPGT contains about 594 amino acids. FPGT has been mapped to chromosomal position 1p31.1.


L-fucose is a key sugar in glycoproteins and other complex carbohydrates since it may be involved in many of the functional roles of these macromolecules, such as in cell-cell recognition. The fucosyl donor for these fucosylated oligosaccharides is GDP-beta-L-fucose. There are two alternate pathways for the biosynthesis of GDP-fucose; the major pathway converts GDP-alpha-D-mannose to GDP-beta-L-fucose. FPGT participates in an alternate pathway that is present in certain mammalian tissues, such as liver and kidney, and appears to function as a salvage pathway to reutilize L-fucose arising from the turnover of glycoproteins and glycolipids. This pathway involves the phosphorylation of L-fucose to form beta-L-fucose-1-phosphate, and then condensation of the beta-L-fucose-1-phosphate with GTP by fucose-1-phosphate guanylyltransferase to form GDP-beta-L-fucose.


GFPP was purified from pig kidney and a partial protein sequence was determined. Human cDNAs encoding a region similar to one of the pig GFPP peptides also were identified. Using a PCR strategy with primers based on one of the ESTs, a cDNA corresponding to the entire human GFPP coding region was cloned. The predicted GFPP protein contains 594 amino acids. When expressed in mammalian cells, the human enzyme exhibited high levels of GFPP activity. Northern blot analysis indicated that the 3.5-kb GFPP mRNA is expressed in several human tissues (Pastuszak et al., GDP-L-fucose pyrophosphorylase: purification, cDNA cloning, and properties of the enzyme. J. Biol. Chem. 273: 30165-30174, 1998).


CARK


CARK is a cardiac-specific kinase gene localized on 1p31.1. Sequence analysis suggests that CARK is a distant family member of integrin-linked kinase. Northern blot and 76-tissue array analyses showed that CARK is highly expressed in heart, but is undetectable in other tissues. Immunohistochemical analysis predominantly localized CARK in the nucleus of cardiac myocytes. In vitro kinase assay showed that CARK is a functional kinase. The yeast two-hybrid system showed that CARK could directly interact with cardiac troponin I, results that were further confirmed by coimmunoprecipitation in vivo (Zhao Y et al. J Mol Med. 2003 May; 81(5): 297-304).


PCLO


The protein PCLO is also known as piccolo (presynaptic cytomatrix protein), ACZ, KIAA0559 and aczonin. PCLO contains about 1225 amino acids. PCLO has been mapped to chromosomal position 7q11.23-q21.1.


Synaptic vesicles dock and fuse in the active zone of the plasma membrane at chemical synapses. The presynaptic cytoskeletal matrix (PCM), which is associated with the active zone and is situated between synaptic vesicles, is thought to be involved in maintaining the neurotransmitter release site in register with the postsynaptic reception apparatus. The cycling of synaptic vesicles is a multistep process involving a number of proteins (see NCBI MIM 603215). Among the components of the PCM that orchestrate these events are Bassoon (BSN; NCBI MIM 604020), RIM (RBBP8; NCBI MIM 604124), Oboe, and Piccolo.


By screening human brain cDNAs for those encoding proteins larger than 50 kD, a partial cDNA encoding PCLO, referred to as KIAA0559, was identified. RT-PCR analysis detected PCLO expression in kidney, with little or no expression in all other tissues tested (Nagase et al., Prediction of the coding sequences of unidentified human genes. IX. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro. DNA Res. 5: 31-39, 1998).


By searching EST and genome databases with a murine Pclo cDNA probe, genomic sequences and a brain-specific EST (KIAA0559) encoding human PCLO were identified. Sequence analysis indicated that the deduced 4,880-amino acid rat Pclo protein is 86% identical to human PCLO. In addition, PCLO shares significant amino acid sequence homology with BSN. BSN and PCLO share 10 homology regions, or PBH regions. PBH1 and PBH2 contain 2 double-zinc finger motifs. PBH4, PBH6, and PBH8 are likely to form coiled-coil structures. At the C terminus, unlike BSN but like RIM and Oboe, PCLO contains a PDZ domain and a C2 domain. The PCLO C2 domain contains all the asp residues required for calcium binding. PCLO also contains multiple proline-rich segments. Confocal microscopy analysis of cultured hippocampal neurons showed colocalization of BSN and PCLO at identical GABAergic and glutamergic synapses, of synaptotagmin (see SYT1; NCBI MIM 185605) and PCLO along dendritic profiles, and of PCLO zinc fingers and PRA1 (NCBI MIM 604925) at nerve terminals (Fenster et al., Piccolo, a presynaptic zinc finger protein structurally related to Bassoon. Neuron 25: 203-214, 2000).


Using cAMP-GEFII (NCBI MIM 606058) as bait in a yeast 2-hybrid screen, mouse piccolo was cloned from an insulin-secreting cell line cDNA library. Northern blot analysis of mouse tissues revealed high levels in cerebrum and cerebellum and moderate levels in pituitary gland, pancreatic islets, and a pheochromocytoma-derived mouse cell line. In situ hybridization of mouse brain revealed piccolo mRNA expressed in cerebral cortex, hippocampus, olfactory bulb, cerebellar cortex, and pituitary gland. The distribution of piccolo mRNA largely overlapped that of cAMP-GEFII and Rim2 (NCBI MIM 606630) mRNA in tissues, cell lines, and mouse brain. Mouse piccolo interacts with both cAMP-GefI and Rim2. In the presence of Ca(2+), the C2A domain of piccolo could homodimerize, it could interact with the C2A domain of Rim2, or it could bind the cAMP GefII-Rim2 complex. It did not bind cAMP-GefII directly. Treatment of pancreatic islets with antisense piccolo oligonucleotides inhibited insulin secretion induced by a cAMP analog and high glucose stimulation. Piccolo serves as a Ca(2+) sensor in exocytosis in pancreatic beta cells and that the formation of a cAMP-GEFII-RIM2 piccolo complex is required (Fujimoto et al., Piccolo, a Ca(2+) sensor in pancreatic beta-cells: involvement of cAMP-GEFII-Rim2-piccolo complex in cAMP-dependent exocytosis. J. Biol. Chem. 277: 50497-50502, 2002).


By comparing the human PCLO genomic sequence with the rat Pclo cDNA, it was determined that the human PCLO gene contains at least 19 exons and spans over 350 kb.


REPS2


The protein REPS2 (RALBP1 associated Eps domain containing 2) is also known as POB1 and partner of Ral-binding protein 1. REPS2 contains about 521 amino acids. REPS2 has been mapped to chromosomal position Xp22.22.


Small G proteins have GDP-bound inactive and GTP-bound active forms; RAL proteins (e.g., RALA; NCBI MIM 179550) shift from the inactive to the active state through the actions of RALGDS (NCBI MIM 601619). RALGDS interacts with the active form of RAS (see HRAS; NCBI MIM 190020).


Using RALA-binding protein-1 (RALBP1; NCBI MIM 605801) as bait in a yeast 2-hybrid screen of a brain cDNA library, cDNAs encoding REP S2, which is designated POB 1, were identified (Ikeda et al., Identification and characterization of a novel protein interacting with Ral-binding protein 1, a putative effector protein of Ral. J. Biol. Chem. 273: 814-821, 1998). Sequence analysis predicted that the 521-amino acid protein has 2 potential initiator methionines in its N terminus, a central EPS 15 (NCBI MIM 600051)-like domain, and 2 proline-rich regions and a putative coiled-coil structure in its C terminus. Northern blot analysis revealed strong expression in rat cerebrum, cerebellum, lung, and testis, with weak expression in kidney and no expression in heart, thymus, liver, spleen, or adrenal gland. Immunoprecipitation and immunoblot analyses confirmed that the C-terminal 146 amino acids of REPS2 and the C-terminal 147 residues of RALBP 1 interact in intact cells. RAL interacts with a distinct region of RALBP 1, just N terminal of the REPS2-binding domain, and both proteins can interact simultaneously with RALBP1. Immunoblot analysis established that REPS2 is tyrosine phosphorylated in response to epidermal growth factor (EGF; NCBI MIM 131530) and interacts with the EGF receptor (EGFR; NCBI MIM 131550), possibly through the adaptor protein GRB2 (NCBI MIM 108355), with which REPS2 interacts specifically.


Using nuclear magnetic resonance spectroscopy, it was shown that the EPS 15 homology domain of REPS2 consists of 2 EF-hand structures, the second of which binds calcium (Koshiba et al., Solution structure of the Eps 15 homology domain of a human POB1 (partner of RalBP1). FEBS Lett. 442: 138-142, 1999).


As used herein, “polymorphic site” refers to a region in a nucleic acid at which two or more alternative nucleotide sequences are observed in a significant number of nucleic acid samples from a population of individuals. A polymorphic site may be a nucleotide sequence of two or more nucleotides, an inserted nucleotide or nucleotide sequence, a deleted nucleotide or nucleotide sequence, or a microsatellite, for example. A polymorphic site that is two or more nucleotides in length may be 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more, 20 or more, 30 or more, 50 or more, 75 or more, 100 or more, 500 or more, or about 1000 nucleotides in length, where all or some of the nucleotide sequences differ within the region. A polymorphic site is often one nucleotide in length, which is referred to herein as a “single nucleotide polymorphism” or a “SNP.”


Where there are two, three, or four alternative nucleotide sequences at a polymorphic site, each nucleotide sequence is referred to as a “polymorphic variant.” Where two polymorphic variants exist, for example, the polymorphic variant represented in aminority of samples from a population is sometimes referred to as a “minor allele” and the polymorphic variant that is more prevalently represented is sometimes referred to as a “major allele.” Many organisms possess a copy of each chromosome (e.g., humans), and those individuals who possess two major alleles or two minor alleles are often referred to as being “homozygous” with respect to the polymorphism, and those individuals who possess one major allele and one minor allele are normally referred to as being “heterozygous” with respect to the polymorphism. Individuals who are homozygous with respect to one allele are sometimes predisposed to a different phenotype as compared to individuals who are heterozygous or homozygous with respect to another allele. As shown hereafter, certain polymorphic variants of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequences are associated with melanoma.


A genotype or polymorphic variant may be expressed in terms of a “haplotype,” which as used herein refers to two or more polymorphic variants occurring within genomic DNA in a group of individuals within a population. For example, two SNPs may exist within a gene where each SNP position includes a cytosine variation and an adenine variation. Certain individuals in a population may carry one allele (heterozygous) or two alleles (homozygous) having the gene with a cytosine at each SNP position. As the two cytosines corresponding to each SNP in the gene travel together on one or both alleles in these individuals, the individuals can be characterized as having a cytosine/cytosine haplotype with respect to the two SNPs in the gene.


As used herein, “phenotype” refers to a trait which can be compared between individuals, such as presence or absence of a condition, a visually observable difference in appearance between individuals, metabolic variations, physiological variations, variations in the function of biological molecules, and the like. An example of a phenotype is occurrence of melanoma.


Researchers sometimes report a polymorphic variant in a database without determining whether the variant is represented in a significant fraction of a population. Because a subset of these reported polymorphic variants are not represented in a statistically significant portion of the population, some of them are sequencing errors and/or not biologically relevant. Thus, it often is not known whether a reported polymorphic variant is statistically significant or biologically relevant until the presence of the variant is detected in a population of individuals and the frequency of the variant is determined. Methods for detecting a polymorphic variant in a population are described herein, such as in Example 2. A polymorphic variant is statistically significant and often biologically relevant if it is represented in 5% or more of a population, sometimes 10% or more, 15% or more, or 20% or more of a population, and often 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more of a population.


A polymorphic variant may be detected on either or both strands of a double-stranded nucleic acid. Also, a polymorphic variant may be located within an intron or exon of a gene or within a portion of a regulatory region such as a promoter, a 5′ untranslated region (UTR), a 3′ UTR, and in DNA (e.g., genomic DNA (gDNA) and complementary DNA (cDNA)), RNA (e.g., mRNA, tRNA, and rRNA), or a polypeptide. Polymorphic variations may or may not result in detectable differences in gene expression, polypeptide structure, or polypeptide function.


For duplex DNA, a polymorphic variation may be reported from one strand or its complementary strand. For example, a thymine at a particular position in SEQ ID NO: 1-17 can be reported as an adenine from the complementary strand. Also, while polymorphic variations at all positions within a haplotype often are reported from the same strand orientation, polymorphic variations at certain positions within a haplotype sometimes are reported from one strand orientation while others are reported from the other. The latter sometimes occurs even though it is understood by the person of ordinary skill in the art that polymorphic variants in a haplotype occur within one strand in a nucleic acid. Where a haplotype is reported from mixed strand orientations, a person of ordinary skill in the art can determine the orientation of each polymorphic variation in the haplotype by analyzing the orientation of each extension oligonucleotide utilized to identify each polymorphic variation.


In genetic analyses that associated polymorphic variations in NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 with melanoma, samples from individuals having melanoma and individuals not having cancer were allelotyped and genotyped. The term “allelotype” as used herein refers to a process for determining the allele frequency for a polymorphic variant in pooled DNA samples from cases and controls. By pooling DNA from each group, an allele frequency for each SNP in each group is calculated. These allele frequencies are then compared to one another. Particular SNPs are considered as being associated with a particular disease when allele frequency differences calculated between case and control pools are statistically significant. The terms “genotyped” and “genotyping” as used herein refer to a process for determining a genotype of one or more individuals, where a “genotype” is a representation of one or more polymorphic variants in a nucleic acid sample from an individual or a population. It was determined that SNPs associated with an increased risk of melanoma existed in NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequences at rs1360457, rs753050, rs729647, rs8404, rs2034453, rs1044639 and rs1904528. Another polymorphic variation associated with an increased risk of melanoma was identified at rs3742536, which coded for a non-synonymous amino acid change at amino acid position 656 in a NID2 amino acid sequence (a proline or serine is encoded at this amino acid position with serine being associated with an increased risk of melanoma).


Polymorphic variants in and around the NRP1 region were tested for association with melanoma. These include polymorphic variants at positions selected from the group consisting of rs7910405, rs5784331, rs6481845, rs2065364, rs10643659, rs7896887, rs7086456, rs6481846, rs7475391, rs7477281, rs7475393, rs7475394, rs7477282, rs7475396, rs2768420, rs9418057, rs7070290, rs1360457, rs1411924, rs5784335, rs2776925, rs2776924, rs1952983, rs869636, rs869637, rs2804470, rs2776928, rs999966, rs2804475, rs4934858, rs4934863, rs4934864, rs1331326, rs1331325, rs4934871, rs4277057, rs2776930, rs1331324, rs2776932, rs4934896, rs2776933, rs4934901, rs1015300, rs2776934, rs2776935, rs2776936, rs2804491, rs2804492, rs2804493, rs2776937, rs2804494, rs4934914, rs2776922, rs2804495, rs2804496, rs4934597, rs2804497, rs2804498, rs2026319, rs1888684, rs3750733, rs4934927, rs2776946, rs4934599, rs4934934, rs2776944, rs2776943, rs2804886, rs2804900, rs2804902, rs2768397, rs2768399, rs2776941, rs2804888, rs2804499, rs2804889, rs2804891, rs2776940, rs2804897, rs2804896, rs2776939 rs2804894, rs2804501, rs2183715, rs2254822, rs2150686, rs2804893, rs2804502, rs2804892, rs2183714, rs2804503, rs1320485, rs4934603, rs2804890 and rs1854145. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057, rs1360457, rs5784335, rs1331325, rs4934871, rs2776930, rs2776935, rs2776936, rs2804491, rs2804494, rs2776922, rs2804495, rs2804496, rs1888684, rs2804888, rs2804891, rs2776940, rs2804897, rs2183714, and rs1854145. Tables in the Examples section hereafter show particular polymorphic variants indicative of a higher risk of melanoma in males and/or females. In specific embodiments, ppolymorphic variants at positions selected from the group consisting of rs7910405, rs5784331, rs6481845, rs2065364, rs10643659, rs7896887, rs7086456, rs6481846, rs7475391, rs7477281, rs7475393, rs7475394, rs7477282, rs7475396, rs2768420, rs9418057, rs7070290, rs1411924 and rs5784335 were tested for an association with increased risk of melanoma. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057, rs5784335. In particular, the following polymorphic variants were associated with risk of melanoma: a thymine at position rs6481845, a guanine-thymine-thymine-thymine deletion at position rs10643659, a cytosine at position rs7475391, a thymine at position rs7475394, a cytosine at position rs7475396, a thymine at position rs9418057 and a thymine at position rs5784335.


Variants in and around the ENDO180 region were tested for association with increased risk of melanoma. These include polymorphic variants at positions selected from the group consisting of rs2429393, rs2429392, rs2465421, rs2465410, rs2460302, rs2460301, rs2465459, rs2429391, rs2014055, rs2252814, rs8079582, rs2429390, rs7209331, rs7405815, rs4968706, rs4968613, rs729647, rs8072984, rs9896444, rs4968617, rs7350907, rs2429389, rs2460300, rs2465454, rs2465452, rs7219711, rs2465440, rs2429388, rs3786130, rs2460289, rs2465428, rs740698, rs2465425, rs2460290, rs3826537, rs3169756, rs1558237, rs2429385, rs5821368, rs8072538, rs2251393, rs2251391, rs2068657, rs8072519, rs7220505, rs2460288, rs4968736, position 16727 in SEQ ID NO: 3. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907, rs8072519. Tables in the Examples section hereafter show particular polymorphic variants indicative of a higher risk of melanoma in males and/or females. In particular, the following polymorphic variants were associated with risk of melanoma: a cytosine at position rs2429390, an adenine at position rs7209331, a cytosine at position rs4968706, a guanine at position rs729647, a guanine at position rs8072984, a thymine at position rs9896444, a cytosine at position rs4968617, a guanine at position rs7350907, a thymine at position rs8072519.


Polymorphic variants in and around the CDK10 region were tested for association with melanoma. These include polymorphic variants at positions selected from the group consisting of rs460879, rs460984, rs2437957, rs3815949, rs2437956, rs2437955, rs2434872, rs467357, rs258337, rs258336, rs258335, rs164752, rs154663, rs164753, rs258332, rs2010623, rs187283, rs258330, rs258328, rs459920, rs166297, rs258318, rs258317, rs171805, rs2081984, rs464586, rs460319, rs447735, rs2377058, rs604868, rs2118193, rs467035, rs2115401, rs417323, rs2377233, rs9746114, rs3751700, rs2277905, rs397891, rs3794638, rs3794637, rs258324, rs258323, rs2075880, rs258322, rs258321, rs164744, rs3833829, rs164743, rs164742, rs187282, rs465507, rs2162943, rs1946482, rs8404, rs1045814, rs4247353, rs462769, rs3803690, rs3751696, rs9972864, rs10567226, rs417414, rs154661, rs4785704, rs4785584, rs4785585, rs4785705, rs4785586, rs4785587, rs2016968, rs3751693, rs3809646, rs4785590, rs2099105, rs4785708, rs4785710, rs4785591, rs4785592, rs4785593, rs4785711, rs4785712, rs4785713, rs4785714, rs4785594, rs3803689, rs4785715, rs3764258, rs4785716, rs4785717, rs2077001, rs2377049, rs2003522, rs1230, rs1800359, rs1061646, rs1800358, rs2286392, rs2074904 and rs2074903. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs460879, rs460984, rs3815949, rs2437955, rs258337, rs164752, rs164753, rs258318, rs464586, rs460319, rs2377233, rs2075880, rs258322, rs258321, rs164744, rs164743, rs164742, rs8404, rs417414, rs4785708, rs4785592, rs4785714, rs4785716 and rs1800359. Tables in the Examples section hereafter show particular polymorphic variants indicative of a higher risk of melanoma in males and/or females. In specific embodiments, polymorphic variants at positions selected from the group consisting of rs9746114, rs3833829, rs8404, rs1045814, rs3751696, rs9972864, rs10567226, position 36589 in SEQ ID NO: 4, position 44544 in SEQ ID NO: 4 and position 48304 in SEQ ID NO: 4. A polymorphic variant position rs8404, especially a cytosine at this position, was associated with an increased risk of melanoma.


Polymorphic variants in and around the FPGT and CARK regions were tested for association with melanoma. These include polymorphic variants at positions selected from the group consisting of rs1412825, rs4422957, rs3835390, rs944795, rs792310, rs526736, rs9804006, rs577367, rs575754, rs573721, rs487917, rs792307, rs476350, rs9425023, rs9425024, rs571848, rs489941, rs491623, rs956, rs520806, rs522759, rs545664, rs545721, rs536355, rs553044, rs581353, rs496008, rs7366854, rs560808, rs485929, rs477134, rs505634, rs542136, rs545680, rs522042, rs575961, rs792321, rs524306, rs495027, rs525358, rs471496, rs504866, rs532396, rs533371, rs567060, rs4650251, rs3753183, rs518769, rs474215, rs500203, rs532221, rs539534, rs531412, rs9970514, rs568049, rs9727250, rs792323, rs3765651, rs3765654, rs481387, rs483259, rs2034453, rs7536997, rs792324, rs6660350, rs492302, rs10493539, rs496720, rs570631, rs4593767, rs576802, rs518574, rs524252, rs792327, rs792328, rs2039407, rs503770, rs503904, rs514012, rs1412827, rs473834, rs480267, rs572180, rs1333029, rs485414, rs1412823, rs2027013, rs548881 and rs792329. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs577367, rs487917, rs792307, rs489941, rs491623, rs536355, rs496008, rs560808, rs542136, rs524306, rs495027, rs504866, rs4650251, rs3753183, rs474215, rs500203, rs532221, rs531412, rs3765654, rs481387, rs483259, rs2034453, rs6660350, rs2039407, rs473834 and rs2027013. Tables in the Examples section hereafter show particular polymorphic variants indicative of a higher risk of melanoma in males and/or females. In specific embodiments, polymorphic variants at positions selected from the group consisting of rs3835390, rs9804006, rs9425023, rs9425024, rs489941, rs496008, rs7366854, rs4650251, rs539534, rs531412, rs9970514, rs568049, rs9727250, rs3765654, rs7536997, rs6660350, rs10493539, rs496720, position 206 in SEQ ID NO: 5, position 9261 in SEQ ID NO: 5, position 37592 in SEQ ID NO: 5. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350, and position 206 in SEQ ID NO: 5. In particular, the following polymorphic variants were associated with risk of melanoma: an adenine at position rs489941, a thymine at position rs496008, an adenine at position rs4650251, an adenine at position rs531412, a cytosine at position rs3765654, a cytosine at position rs6660350, and a guanine at position 206 in SEQ ID NO: 5.


Polymorphic variants in and around the PCLO region were tested for association with melanoma. These include polymorphic variants at positions selected from the group consisting of rs2158220, rs1024465, rs4732483, rs4732485, rs1859176, rs1859177, rs2074401, rs4299948, rs4348435, rs4348436, rs4413718, rs985199, rs7791413, rs2522830, rs2214414, rs2189246, rs5885312, rs3064840, rs2051791, rs6467914, rs2214415, rs2189247, rs3064869, rs2522831, rs2715147, rs3064870, rs2715148, rs2522832, rs1044639, rs2715149, rs2522833, rs2247523, rs2371214, rs2189248, rs2715150, rs2715151, rs2371215, rs2715152, rs2715153, rs972346, rs2715154, rs2522834, rs2023847, rs2715155, rs2715156, rs10715043, rs2301722, rs2257207, rs2715157, rs2715158, rs2715159, rs2522835, rs2522836, rs2522837, rs2522838, rs2522839, rs2522840, rs2522841, rs10555175, rs10592258, rs2522842, rs6956848, rs2522843, rs2522844, rs2522845, rs2715161, rs6972720, rs4377905, rs4579453, rs4260846, rs1986481, rs4628206 and rs2888017. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs1859176, rs2522830, rs2214414, rs2051791, rs6467914, rs2189247, rs3064869, rs2715148, rs1044639, rs2522833, rs2371214, rs2715151, rs2371215, rs2715153, rs2715154, rs2023847, rs2715155, rs2715156, rs2715157, rs2715158, rs2715159, rs2522835, rs2522837, rs10555175, rs6956848, rs2522845, rs4579453 and rs1986481. Tables in the Examples section hereafter show particular polymorphic variants indicative of a higher risk of melanoma in males and/or females. In specific embodiments, polymorphic variants at positions selected from the group consisting of rs7791413, rs2189246, rs5885312, rs3064840, rs6467914, rs3064869, rs3064870, rs10715043, rs2522841, rs10555175, rs10592258, rs6956848, rs6972720, position 53843 in SEQ ID NO: 6, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6. In particular, the following polymorphic variants were associated with risk of melanoma: a thymine at position rs6467914, an adenine at position rs3064869, a thymine at position rs10555175, a guanine at position rs6956848, position in SEQ ID NO: 6 selected from the group consisting of a guanine at position 63624 and an adenine-thymine insert at position 65039.


Polymorphic variants in and around the REPS2 region were tested for association with melanoma. These include polymorphic variants at positions selected from the group consisting of rs1904525, rs5924557, rs5924616, rs5924617, rs7883612, rs5924618, rs5924619, rs5901596, rs7878645, rs5924564, rs5924535, rs6629197, rs7060904, rs4828523, rs5924568, rs5924569, rs1904528, rs5924570, rs1904527, rs2052763, rs1549529, rs5924573, rs5901600, rs2288304, rs4240072, rs3055191, rs6632943, rs5924580, rs4828544, rs1807412, rs2010456, rs5969758, rs1965051, rs9887218, rs1426800, rs5969759, rs6527766, rs1365529, rs5924582, rs2114458, rs5924583, rs5924545, rs5969761, rs6629201, rs6629202, rs5924584, rs5924585, rs933946, rs5969762, rs713355, rs1365530, rs1961490, rs6527769, rs2006847, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs1426801, rs5924546, rs5924547, rs1365528, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924591, rs5924592, rs5924548, rs5924593, rs4828547, rs714070, rs5969768, rs4828526, rs5969738, rs5924595, rs5924549, rs4828528, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs7891150, rs4828550, rs5924603, rs1807947, rs954253, rs2382814, rs1896178, rs1896177, rs5924605, rs5924606, position 10537 in SEQ ID NO: 7, position 25216 in SEQ ID NO: 7 and position 119368 in SEQ ID NO: 7. Polymorphic variants at the following positions in particular were associated with an increased risk of melanoma: rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606, and position 25216 in SEQ ID NO: 7. Tables in the Examples section hereafter show particular polymorphic variants indicative of a higher risk of melanoma in males and/or females. In particular, the following polymorphic variants were associated with risk of melanoma: a thymine at position rs5924619, a cytosine at position rs5924564, a guanine at position rs4828523, an adenine at position rs5924569, a guanine at position rs2052763, an adenine at position rs1549529, an adenine at position rs5924573, an adenine at position rs4240072, a guanine at position rs5924580, a cytosine at position rs4828544, a guanine at position rs5969758, an adenine at position rs1965051, an adenine at position rs1426800, a cytosine at position rs1365529, an adenine at position rs5924545, a thymine at position rs5969761, a thymine at position rs6629202, a guanine at position rs5924584, a cytosine at position rs1365530, a cytosine at position rs6527769, a guanine at position rs2382813, a guanine at position rs2033120, an adenine at position rs2162394, a cytosine at position rs1346246, a cytosine at position rs1365527, an adenine at position rs4828524, a thymine at position rs5924588, a thymine at position rs5969764, a thymine at position rs5924589, a thymine at position rs5924590, a guanine at position rs5924593, a guanine at position rs4828547, a cytosine at position rs5969768, a guanine at position rs4828526, an adenine at position rs5924595, a guanine at position rs5924549, an adenine at position rs5969769, a thymine at position rs4007744, a thymine at position rs2891073, a guanine at position rs5924599, an adenine at position rs1549531, a guanine at position rs5924601, a cytosine at position rs5901609, a cytosine at position rs5969739, a thymine at position rs5924602, an adenine at position rs1426798, an adenine at position rs4828550, a thymine at position rs5924603, an adenine at position rs1807947, a thymine at position rs2382814, an adenine at position rs5924606, and an adenine at position 25216 in SEQ ID NO: 7.


The embodiments described hereafter may be directed to any of the polymorphic variants described herein. Sometimes, embodiments are directed to any of the polymorphic variants described herein with the proviso that the embodiments are not directed to a polymorphic variant at one or more positions selected from the group consisting of rs1360457, rs753050, rs729647, rs8404, rs2034453, rs1044639 and rs1904528.


Additional Polymorphic Variants Associated with Melanoma


Also provided is a method for identifying polymorphic variants proximal to an incident, founder polymorphic variant associated with melanoma. Thus, featured herein are methods for identifying a polymorphic variation associated with melanoma that is proximal to an incident polymorphic variation associated with melanoma, which comprises identifying a polymorphic variant proximal to the incident polymorphic variant associated with melanoma, where the incident polymorphic variant is in a nucleotide sequence set forth in SEQ ID NO: 1-17. The nucleotide sequence often comprises a polynucleotide sequence selected from the group consisting of (a) a polynucleotide sequence set forth in SEQ ID NO: 1-17; (b) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence encoded by a nucleotide sequence set forth in SEQ ID NO: 1-17; and (c) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence that is 90% or more identical to an amino acid sequence encoded by a nucleotide sequence set forth in SEQ ID NO: 1-17 or a polynucleotide sequence 90% or more identical to the polynucleotide sequence set forth in SEQ ID NO: 1-17. The presence or absence of an association of the proximal polymorphic variant with NIDDM then is determined using a known association method, such as a method described in the Examples hereafter. In an embodiment, the incident polymorphic variant is described in SEQ ID NO: 1-17 or in the Examples below. In another embodiment, the proximal polymorphic variant identified sometimes is a publicly disclosed polymorphic variant, which for example, sometimes is published in a publicly available database. In other embodiments, the polymorphic variant identified is not publicly disclosed and is discovered using a known method, including, but not limited to, sequencing a region surrounding the incident polymorphic variant in a group of nucleic samples. Thus, multiple polymorphic variants proximal to an incident polymorphic variant are associated with melanoma using this method.


The proximal polymorphic variant often is identified in a region surrounding the incident polymorphic variant. In certain embodiments, this surrounding region is about 50 kb flanking the first polymorphic variant (e.g about 50 kb 5′ of the first polymorphic variant and about 50 kb 3′ of the first polymorphic variant), and the region sometimes is composed of shorter flanking sequences, such as flanking sequences of about 40 kb, about 30 kb, about 25 kb, about 20 kb, about 15 kb, about 10 kb, about 7 kb, about 5 kb, or about 2 kb 5′ and 3′ of the incident polymorphic variant. In other embodiments, the region is composed of longer flanking sequences, such as flanking sequences of about 55 kb, about 60 kb, about 65 kb, about 70 kb, about 75 kb, about 80 kb, about 85 kb, about 90 kb, about 95 kb, or about 100 kb 5′ and 3′ of the incident polymorphic variant.


In certain embodiments, polymorphic variants associated with melanoma are identified iteratively. For example, a first proximal polymorphic variant is associated with melanoma using the methods described above and then another polymorphic variant proximal to the first proximal polymorphic variant is identified (e.g., publicly disclosed or discovered) and the presence or absence of an association of one or more other polymorphic variants proximal to the first proximal polymorphic variant with melanoma is determined.


The methods described herein are useful for identifying or discovering additional polymorphic variants that may be used to further characterize a gene, region or loci associated with a condition, a disease (e.g., melanoma), or a disorder. For example, allelotyping or genotyping data from the additional polymorphic variants may be used to identify a functional mutation or a region of linkage disequilibrium. In certain embodiments, polymorphic variants identified or discovered within a region comprising the first polymorphic variant associated with melanoma are genotyped using the genetic methods and sample selection techniques described herein, and it can be determined whether those polymorphic variants are in linkage disequilibrium with the first polymorphic variant. The size of the region in linkage disequilibrium with the first polymorphic variant also can be assessed using these genotyping methods. Thus, provided herein are methods for determining whether a polymorphic variant is in linkage disequilibrium with a first polymorphic variant associated with melanoma, and such information can be used in prognosis methods described herein.


Isolated Nucleic Acids and Variants Thereof


Featured herein are isolated NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids, which include the nucleotide sequence of SEQ ID NO: 1-17, NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid variants, and substantially identical nucleic acids and fragments of the foregoing. Nucleotide sequences of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids sometimes are referred to herein as “NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequences.” A “NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid variant” refers to one allele that may have one or more different polymorphic variations as compared to another allele in another subject or the same subject. A polymorphic variation in the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid variant may be represented on one or both strands in a double-stranded nucleic acid or on one chromosomal complement (heterozygous) or both chromosomal complements (homozygous). A NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid may comprise one or more polymorphic variations associated with melanoma described herein.


As used herein, the term “nucleic acid” includes DNA molecules (e.g., a complementary DNA (cDNA) and genomic DNA (gDNA)) and RNA molecules (e.g., mRNA, rRNA, siRNA and tRNA) and analogs of DNA or RNA, for example, by use of nucleotide analogs. The nucleic acid molecule can be single-stranded and it is often double-stranded. The term “isolated or purified nucleic acid” refers to nucleic acids that are separated from other nucleic acids present in the natural source of the nucleic acid. For example, with regard to genomic DNA, the term “isolated” includes nucleic acids which are separated from the chromosome with which the genomic DNA is naturally associated. An “isolated” nucleic acid is often free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′ and/or 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of 5′ and/or 3′ nucleotide sequences which flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. As used herein, the term “NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene” refers to a nucleotide sequence that encodes a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide.


In particular embodiments, a nucleic acid comprises one or more polymorphic variations independently selected from the group consisting of an adenine at position 16727 in SEQ ID NO: 3, a guanine at position 36589 in SEQ ID NO: 4, a thymine at position 44544 in SEQ ID NO: 4, a guanine at position 48304 in SEQ ID NO: 4, a guanine at position 53843 in SEQ ID NO: 6, a guanine at position 63624 in SEQ ID NO: 6, an adenine-thymine deletion at position 65039 in SEQ ID NO: 6, a guanine at position 206 in SEQ ID NO: 5, an adenine at position 9261 in SEQ ID NO: 5, a thymine at position 37592 in SEQ ID NO: 5, a guanine at position 10537 in SEQ ID NO: 7, a cytosine at position 25216 in SEQ ID NO: 7 and a thymine at position 119368 in SEQ ID NO: 7. The nucleic acid often comprises a part of or all of a nucleotide sequence in SEQ ID NO: 3, 4, 5, 6 and/or 7, and sometimes such a nucleotide sequence is a 5′ and/or 3′ sequence flanking a polymorphic variant described above that is 5, 6, 7 . . . 50, 51, 52 . . . 100, 101, 102 . . . 500, 501, 502 . . . 999 or 1000 nucleotides in length. Other embodiments are directed to methods of identifying a polymorphic variation at one or more positions in a nucleic acid (e.g., genotyping at one or more positions in the nucleic acid), where the one or more positions are independently selected from the group consisting of position 16727 in SEQ ID NO: 3, position 36589 in SEQ ID NO: 4, position 44544 in SEQ ID NO: 4, position 48304 in SEQ ID NO: 4, position 53843 in SEQ ID NO: 6, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6, position 206 in SEQ ID NO: 5, position 9261 in SEQ ID NO: 5, position 37592 in SEQ ID NO: 5, position 10537 in SEQ ID NO: 7, position 25216 in SEQ ID NO: 7 and position 119368 in SEQ ID NO: 7.


Also included herein are nucleic acid fragments. These fragments are typically a nucleotide sequence identical to a nucleotide sequence in SEQ ID NO: 1-17, a nucleotide sequence substantially identical to a nucleotide sequence in SEQ ID NO: 1-17, or a nucleotide sequence that is complementary to the foregoing. The nucleic acid fragment may be identical, substantially identical or homologous to a nucleotide sequence in an exon or an intron in SEQ ID NO: 1-17 and may encode a full-length or mature polypeptide, or may encode a domain or part of a domain of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Sometimes, the fragment will comprises one or more of the polymorphic variations described herein as being associated with melanoma. The nucleic acid fragment is often 50, 100, or 200 or fewer base pairs in length, and is sometimes about 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, or 1400 base pairs in length. A nucleic acid fragment that is complementary to a nucleotide sequence identical or substantially identical to the nucleotide sequence of SEQ ID NO: 1-17 and hybridizes to such a nucleotide sequence under stringent conditions is often referred to as a “probe.” Nucleic acid fragments often include one or more polymorphic sites, or sometimes have an end that is adjacent to a polymorphic site as described hereafter.


An example of a nucleic acid fragment is an oligonucleotide. As used herein, the term “oligonucleotide” refers to a nucleic acid comprising about 8 to about 50 covalently linked nucleotides, often comprising from about 8 to about 35 nucleotides, and more often from about 10 to about 25 nucleotides. The backbone and nucleotides within an oligonucleotide may be the same as those of naturally occurring nucleic acids, or analogs or derivatives of naturally occurring nucleic acids, provided that oligonucleotides having such analogs or derivatives retain the ability to hybridize specifically to a nucleic acid comprising a targeted polymorphism. Oligonucleotides described herein may be used as hybridization probes or as components of prognostic or diagnostic assays, for example, as described herein.


Oligonucleotides are typically synthesized using standard methods and equipment, such as the ABI™3900 High Throughput DNA Synthesizer and the EXPEDITE™ 8909 Nucleic Acid Synthesizer, both of which are available from Applied Biosystems (Foster City, Calif.). Analogs and derivatives are exemplified in U.S. Pat. Nos. 4,469,863; 5,536,821; 5,541,306; 5,637,683; 5,637,684; 5,700,922; 5,717,083; 5,719,262; 5,739,308; 5,773,601; 5,886,165; 5,929,226; 5,977,296; 6,140,482; WO 00/56746; WO 01/14398, and related publications. Methods for synthesizing oligonucleotides comprising such analogs or derivatives are disclosed, for example, in the patent publications cited above and in U.S. Pat. Nos. 5,614,622; 5,739,314; 5,955,599; 5,962,674; 6,117,992; in WO 00/75372; and in related publications.


Oligonucleotides may also be linked to a second moiety. The second moiety may be an additional nucleotide sequence such as a tail sequence (e.g., a polyadenosine tail), an adapter sequence (e.g., phage M13 universal tail sequence), and others. Alternatively, the second moiety may be a non-nucleotide moiety such as a moiety which facilitates linkage to a solid support or a label to facilitate detection of the oligonucleotide. Such labels include, without limitation, a radioactive label, a fluorescent label, a chemiluminescent label, a paramagnetic label, and the like. The second moiety may be attached to any position of the oligonucleotide, provided the oligonucleotide can hybridize to the nucleic acid comprising the polymorphism.


Uses for Nucleic Acids


Nucleic acid coding sequences depicted in SEQ ID NO: 1-17, or substantially identical sequences thereof, may be used for diagnostic purposes for detection and control of polypeptide expression. Also, included are oligonucleotide sequences such as antisense nucleic acids (e.g., DNA, RNA or PNA), inhibitory RNA and small-interfering RNA (siRNA), and ribozymes that function to inhibit translation of a polypeptide. Antisense techniques and RNA interference techniques are known in the art and are described herein.


Ribozymes are enzymatic RNA molecules capable of catalyzing the specific cleavage of RNA. The mechanism of ribozyme action involves sequence specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. For example, hammerhead motif ribozyme molecules may be engineered that specifically and efficiently catalyze endonucleolytic cleavage of RNA sequences encoded by a nucleotide sequence set forth in SEQ ID NO: 1-17 or a substantially identical sequence thereof. Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites which include the following sequences, GUA, GUU and GUC. Once identified, short RNA sequences of between fifteen (15) and twenty (20) ribonucleotides corresponding to the region of the target gene containing the cleavage site may be evaluated for predicted structural features such as secondary structure that may render the oligonucleotide sequence unsuitable. The suitability of candidate targets may also be evaluated by testing their accessibility to hybridization with complementary oligonucleotides, using ribonuclease protection assays.


Antisense RNA and DNA molecules, siRNA and ribozymes may be prepared by any method known in the art for the synthesis of RNA molecules. These include techniques for chemically synthesizing oligodeoxyribonucleotides well known in the art such as solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding the antisense RNA molecule. Such DNA sequences may be incorporated into a wide variety of vectors which incorporate suitable RNA polymerase promoters such as the T7 or SP6 polymerase promoters. Alternatively, antisense cDNA constructs that synthesize antisense RNA constitutively or inducibly, depending on the promoter used, can be introduced stably into cell lines.


DNA encoding a polypeptide also may have a number of uses for the diagnosis of diseases, including melanoma, resulting from aberrant expression of a target gene described herein. For example, the nucleic acid sequence may be used in hybridization assays of biopsies or autopsies to diagnose abnormalities of expression or function (e.g., Southern or Northern blot analysis, in situ hybridization assays).


In addition, the expression of a polypeptide during embryonic development may also be determined using nucleic acid encoding the polypeptide. As addressed, infra, production of functionally impaired polypeptide is the cause of various disease states, melanoma. In situ hybridizations using polypeptide as a probe may be employed to predict problems related to melanoma. Further, as indicated, infra, administration of human active polypeptide, recombinantly produced as described herein, may be used to treat disease states related to functionally impaired polypeptide. Alternatively, gene therapy approaches may be employed to remedy deficiencies of functional polypeptide or to replace or compete with dysfunctional polypeptide.


Expression Vectors, Host Cells, and Genetically Engineered Cells


Provided herein are nucleic acid vectors, often expression vectors, which contain a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid. As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked and can include a plasmid, cosmid, or viral vector. The vector can be capable of autonomous replication or it can integrate into a host DNA. Viral vectors may include replication defective retroviruses, adenoviruses and adeno-associated viruses for example.


A vector can include a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid in a form suitable for expression of the nucleic acid in a host cell. The recombinant expression vector typically includes one or more regulatory sequences operatively linked to the nucleic acid sequence to be expressed. The term “regulatory sequence” includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Regulatory sequences include those that direct constitutive expression of a nucleotide sequence, as well as tissue-specific regulatory and/or inducible sequences. The design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of polypeptide desired, and the like. Expression vectors can be introduced into host cells to produce NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides, including fusion polypeptides, encoded by NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids.


Recombinant expression vectors can be designed for expression of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides in prokaryotic or eukaryotic cells. For example, NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides can be expressed in E. coli, insect cells (e.g., using baculovirus expression vectors), yeast cells, or mammalian cells. Suitable host cells are discussed further in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.


Expression of polypeptides in prokaryotes is most often carried out in E. coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion polypeptides. Fusion vectors add a number of amino acids to a polypeptide encoded therein, usually to the amino terminus of the recombinant polypeptide. Such fusion vectors typically serve three purposes: 1) to increase expression of recombinant polypeptide; 2) to increase the solubility of the recombinant polypeptide; and 3) to aid in the purification of the recombinant polypeptide by acting as a ligand in affinity purification. Often, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant polypeptide to enable separation of the recombinant polypeptide from the fusion moiety subsequent to purification of the fusion polypeptide. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith & Johnson, Gene 67: 31-40 (1988)), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse glutathione S-transferase (GST), maltose E binding polypeptide, or polypeptide A, respectively, to the target recombinant polypeptide.


Purified fusion polypeptides can be used in screening assays and to generate antibodies specific for NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides. In a therapeutic embodiment, fusion polypeptide expressed in a retroviral expression vector is used to infect bone marrow cells that are subsequently transplanted into irradiated recipients. The pathology of the subject recipient is then examined after sufficient time has passed (e.g., six (6) weeks).


Expressing the polypeptide in host bacteria with an impaired capacity to proteolytically cleave the recombinant polypeptide is often used to maximize recombinant polypeptide expression (Gottesman, S., Gene Expression Technology: Methods in Enzymology, Academic Press, San Diego, Calif. 185: 119-128 (1990)). Another strategy is to alter the nucleotide sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli (Wada et al., Nucleic Acids Res. 20: 2111-2118 (1992)). Such alteration of nucleotide sequences can be carried out by standard DNA synthesis techniques.


When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, Adenovirus 2, cytomegalovirus and Simian Virus 40. Recombinant mammalian expression vectors are often capable of directing expression of the nucleic acid in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Non-limiting examples of suitable tissue-specific promoters include an albumin promoter (liver-specific; Pinkert et al., Genes Dev. 1: 268-277 (1987)), lymphoid-specific promoters (Calame & Eaton, Adv. Immunol. 43: 235-275 (1988)), promoters of T cell receptors (Winoto & Baltimore, EMBO J. 8: 729-733 (1989)) promoters of immunoglobulins (Banerji et al., Cell 33: 729-740 (1983); Queen & Baltimore, Cell 33: 741-748 (1983)), neuron-specific promoters (e.g., the neurofilament promoter; Byrne & Ruddle, Proc. Natl. Acad. Sci. USA 86: 5473-5477 (1989)), pancreas-specific promoters (Edlund et al., Science 230: 912-916 (1985)), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are sometimes utilized, for example, the murine hox promoters (Kessel & Gruss, Science 249: 374-379 (1990)) and the a-fetopolypeptide promoter (Campes & Tilghman, Genes Dev. 3: 537-546 (1989)).


A NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid may also be cloned into an expression vector in an antisense orientation. Regulatory sequences (e.g., viral promoters and/or enhancers) operatively linked to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid cloned in the antisense orientation can be chosen for directing constitutive, tissue specific or cell type specific expression of antisense RNA in a variety of cell types. Antisense expression vectors can be in the form of a recombinant plasmid, phagemid or attenuated virus. For a discussion of the regulation of gene expression using antisense genes see Weintraub et al., Antisense RNA as a molecular tool for genetic analysis, Reviews—Trends in Genetics, Vol. 1(1) (1986).


Also provided herein are host cells that include a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid within a recombinant expression vector or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid sequence fragments which allow it to homologously recombine into a specific site of the host cell genome. The terms “host cell” and “recombinant host cell” are used interchangeably herein. Such terms refer not only to the particular subject cell but rather also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein. A host cell can be any prokaryotic or eukaryotic cell. For example, a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are known to those skilled in the art.


Vectors can be introduced into host cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, transduction/infection, DEAE-dextran-mediated transfection, lipofection, or electroporation.


A host cell provided herein can be used to produce (i.e., express) a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Accordingly, further provided are methods for producing a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide using the host cells described herein. In one embodiment, the method includes culturing host cells into which a recombinant expression vector encoding a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide has been introduced in a suitable medium such that a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide is produced. In another embodiment, the method further includes isolating a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide from the medium or the host cell.


Also provided are cells or purified preparations of cells which include a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 transgene, or which otherwise misexpress NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Cell preparations can consist of human or non-human cells, e.g., rodent cells, e.g., mouse or rat cells, rabbit cells, or pig cells. In embodiments, the cell or cells include a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 transgene (e.g., a heterologous form of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 such as a human gene expressed in non-human cells). The NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 transgene can be misexpressed, e.g., overexpressed or underexpressed. In other embodiments, the cell or cells include a gene which misexpress an endogenous NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide (e.g., expression of a gene is disrupted, also known as a knockout). Such cells can serve as a model for studying disorders which are related to mutated or mis-expressed NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 alleles or for use in drug screening. Also provided are human cells (e.g., a hematopoietic stem cells) transformed with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid.


Also provided are cells or a purified preparation thereof (e.g., human cells) in which an endogenous NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid is under the control of a regulatory sequence that does not normally control the expression of the endogenous NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene. The expression characteristics of an endogenous gene within a cell (e.g., a cell line or microorganism) can be modified by inserting a heterologous DNA regulatory element into the genome of the cell such that the inserted regulatory element is operably linked to the endogenous NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene. For example, an endogenous NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene (e.g., a gene which is “transcriptionally silent,” not normally expressed, or expressed only at very low levels) may be activated by inserting a regulatory element which is capable of promoting the expression of a normally expressed gene product in that cell. Techniques such as targeted homologous recombinations, can be used to insert the heterologous DNA as described in, e.g., Chappel, U.S. Pat. No. 5,272,071; WO 91/06667, published on May 16, 1991.


Transgenic Animals


Non-human transgenic animals that express a heterologous NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide (e.g., expressed from a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid isolated from another organism) can be generated. Such animals are useful for studying the function and/or activity of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide and for identifying and/or evaluating modulators of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid and NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide activity. As used herein, a “transgenic animal” is a non-human animal such as a mammal (e.g., a non-human primate such as chimpanzee, baboon, or macaque; an ungulate such as an equine, bovine, or caprine; or a rodent such as a rat, a mouse, or an Israeli sand rat), a bird (e.g., a chicken or a turkey), an amphibian (e.g., a frog, salamander, or newt), or an insect (e.g., Drosophila melanogaster), in which one or more of the cells of the animal includes a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 transgene. A transgene is exogenous DNA or a rearrangement (e.g., a deletion of endogenous chromosomal DNA) that is often integrated into or occurs in the genome of cells in a transgenic animal. A transgene can direct expression of an encoded gene product in one or more cell types or tissues of the transgenic animal, and other transgenes can reduce expression (e.g., a knockout). Thus, a transgenic animal can be one in which an endogenous NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell of the animal (e.g., an embryonic cell of the animal) prior to development of the animal.


Intronic sequences and polyadenylation signals can also be included in the transgene to increase expression efficiency of the transgene. One or more tissue-specific regulatory sequences can be operably linked to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 transgene to direct expression of a NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide to particular cells. A transgenic founder animal can be identified based upon the presence of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 transgene in its genome and/or expression of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA in tissues or cells of the animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene encoding a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide can further be bred to other transgenic animals carrying other transgenes.


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides can be expressed in transgenic animals or plants by introducing, for example, a nucleic acid encoding the polypeptide into the genome of an animal. In embodiments the nucleic acid is placed under the control of a tissue specific promoter, e.g., a milk or egg specific promoter, and recovered from the milk or eggs produced by the animal. Also included is a population of cells from a transgenic animal.


NRP1. NID2. ENDO180. CDK10. FPGT. CARK, PCLO or REPS2 Polypeptides


Also featured herein are isolated NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides, including proteins and peptides, that include an amino acid sequence set forth in SEQ ID NO: 18-27, or a substantially identical sequence thereof or variant thereof. In an embodiment, an NID2 polypeptide includes a serine at amino acid position 656. Isolated NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides featured herein include both the full-length polypeptide and the mature polypeptide (i.e., the polypeptide minus the signal sequence or propeptide domain). A NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide is a polypeptide encoded by a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, where one nucleic acid can encode one or more different polypeptides. An “isolated” or “purified” polypeptide or protein is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. In one embodiment, the language “substantially free” means preparation of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide variant having less than about 30%, 20%, 10% and more preferably 5% (by dry weight), of non-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide (also referred to herein as a “contaminating protein”), or of chemical precursors or non-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 chemicals. When the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or a biologically active portion thereof is recombinantly produced, it is also preferably substantially free of culture medium, specifically, where culture medium represents less than about 20%, sometimes less than about 10%, and often less than about 5% of the volume of the polypeptide preparation. Isolated or purified NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide preparations are sometimes 0.01 milligrams or more or 0.1 milligrams or more, and often 1.0 milligrams or more and 10 milligrams or more in dry weight.


Further included herein are NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide fragments. The polypeptide fragment may be a domain or part of a domain of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK PCLO or REPS2 polypeptide. The polypeptide fragment may have increased, decreased or unexpected biological activity. The polypeptide fragment is often 50 or fewer, 100 or fewer, or 200 or fewer amino acids in length, and is sometimes 300, 400, 500, 600, or 700, or fewer amino acids in length. A fragment sometimes includes a substantial portion of a domain, examples of which are described herein.


Substantially identical polypeptides may depart from the amino acid sequences set forth in SEQ ID NO: 18-27 in different manners. For example, conservative amino acid modifications may be introduced at one or more positions in the amino acid sequences of SEQ ID NO: 18-27. A “conservative amino acid substitution” is one in which the amino acid is replaced by another amino acid having a similar structure and/or chemical function. Families of amino acid residues having similar structures and functions are well known. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Also, essential and non-essential amino acids may be replaced. A “non-essential” amino acid is one that can be altered without abolishing or substantially altering the biological function of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, whereas altering an “essential” amino acid abolishes or substantially alters the biological function of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Amino acids that are conserved among NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides are typically essential amino acids.


Also, NRP1, NID2, ENDO180, CDK10, FPGT, CARK PCLO or REPS2 polypeptides and polypeptide variants may exist as chimeric or fusion polypeptides. As used herein, a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 “chimeric polypeptide” or “fusion polypeptide” includes a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide linked to a non-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. A “non-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide” refers to a polypeptide having an amino acid sequence corresponding to a polypeptide which is not substantially identical to the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, which includes, for example, a polypeptide that is different from the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide and derived from the same or a different organism. The NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide in the fusion polypeptide can correspond to an entire or nearly entire NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or a fragment thereof. The non-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide can be fused to the N-terminus or C-terminus of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide.


Fusion polypeptides can include a moiety having high affinity for a ligand. For example, the fusion polypeptide can be a GST-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 fusion polypeptide in which the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 sequences are fused to the C-terminus of the GST sequences, or a polyhistidine-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 fusion polypeptide in which the NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide is fused at the N- or C-terminus to a string of histidine residues. Such fusion polypeptides can facilitate purification of recombinant NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2. Expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide), and a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid can be cloned into an expression vector such that the fusion moiety is linked in-frame to the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Further, the fusion polypeptide can be a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression, secretion, cellular internalization, and cellular localization of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide can be increased through use of a heterologous signal sequence. Fusion polypeptides can also include all or a part of a serum polypeptide (e.g., an IgG constant region or human serum albumin).


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides or fragments thereof can be incorporated into pharmaceutical compositions and administered to a subject in vivo. Administration of these NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides can be used to affect the bioavailability of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 substrate and may effectively increase NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 biological activity in a cell. NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 fusion polypeptides may be useful therapeutically for the treatment of disorders caused by, for example, (i) aberrant modification or mutation of a gene encoding a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide; (ii) mis-regulation of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene; and (iii) aberrant post-translational modification of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Also, NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides can be used as immunogens to produce anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibodies in a subject, to purify NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 ligands or binding partners, and in screening assays to identify molecules which inhibit or enhance the interaction of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 substrate.


In addition, polypeptides can be chemically synthesized using techniques known in the art (See, e.g., Creighton, 1983 Proteins. New York, N.Y.: W. H. Freeman and Company; and Hunkapiller et al., (1984) Nature July 12-18; 310(5973): 105-11). For example, a relative short fragment can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the fragment sequence. Nonclassical amino acids or chemical amino acid analogs include but are not limited to ornithine (hereinafter referred to as Z), diaminobutyric acid (hereinafter referred to as B), norleucine (hereinafter referred to as O), pyrylalanine, thienylalanine, naphthylalanine and phenylglycine. Other examples of non-naturally occurring amino acids and non-classical amino acid replacements are alpha* and alpha-disubstituted* amino acids, N-alkyl amino acids*, lactic acid*, halide derivatives of natural amino acids such as trifluorotyrosine*, p-Cl-phenylalanine*, p-Br-phenylalanine*, p-I-phenylalanine*, L-allyl-glycine*, beta-alanine*, L-alpha-amino butyric acid*, L-gamma-amino butyric acid*, L-alpha-amino isobutyric acid*, L-epsilon-amino caproic acid#, 7-amino heptanoic acid*, L-methionine sulfone*, L-norleucine*, L-norvaline*, p-nitro-L-phenylalanine*, L-hydroxyproline#, L-thioproline*, methyl derivatives of phenylalanine (Phe) such as 4-methyl-Phe*, pentamethyl-Phe*, L-Phe (4-amino)#, L-Tyr (methyl)*, L-Phe (4-isopropyl)*, L-Tic (1,2,3,4-tetrahydroisoquinoline-3-carboxyl acid)*, L-diaminopropionic acid, L-Phe (4-benzyl)*, 2,4-diaminobutyric acid, 4-aminobutyric acid (gamma-Abu), 2-amino butyric acid (alpha-Abu), 6-amino hexanoic acid (epsilon-Ahx), 2-amino isobutyric acid (Aib), 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, fluoroamino acids, designer amino acids such as beta-methyl amino acids, Ca-methyl amino acids, Na-methyl amino acids, naphthyl alanine, amino acid analogs in general and the like. The notation * indicates a derivative having hydrophobic characteristics, # indicates a derivative having hydrophilic characteristics, and #* indicates a derivative having amphipathic characteristics.


Polypeptides and polypeptide fragments sometimes are differentially modified during or after translation, e.g., by glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to an antibody molecule or other cellular ligand, etc. Any of numerous chemical modifications may be carried out by known techniques, including but not limited, to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease, NaBH4; acetylation, formylation, oxidation, reduction; metabolic synthesis in the presence of tunicamycin; and the like. Additional post-translational modifications include, for example, N-linked or O-linked carbohydrate chains, processing of N-terminal or C-terminal ends), attachment of chemical moieties to the amino acid backbone, chemical modifications of N-linked or O-linked carbohydrate chains, and addition or deletion of an N-terminal methionine residue as a result of procaryotic host cell expression. The polypeptide fragments may also be modified with a detectable label, such as an enzymatic, fluorescent, isotopic or affinity label to allow for detection and isolation of the polypeptide.


Also provided are chemically modified derivatives of polypeptides that can provide additional advantages such as increased solubility, stability and circulating time of the polypeptide, or decreased immunogenicity (see e.g., U.S. Pat. No. 4,179,337). The chemical moieties for derivitization may be selected from water soluble polymers such as polyethylene glycol, ethylene glycol/propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol and the like. The polypeptides may be modified at random positions within the molecule, or at predetermined positions within the molecule and may include one, two, three or more attached chemical moieties.


The polymer may be of any molecular weight, and may be branched or unbranched. For polyethylene glycol, the molecular weight often utilized is between about 1 kDa and about 100 kDa (the term “about” indicating that in preparations of polyethylene glycol, some molecules will weigh more, some less, than the stated molecular weight) for ease in handling and manufacturing. Other sizes may be used, depending on the desired therapeutic profile (e.g., the duration of sustained release desired, the effects, if any on biological activity, the ease in handling, the degree or lack of antigenicity and other known effects of the polyethylene glycol to a therapeutic protein or analog).


The polymers should be attached to the polypeptide with consideration of effects on functional or antigenic domains of the polypeptide. There are a number of attachment methods available to those skilled in the art (e.g., EP 0 401 384 (coupling PEG to G-CSF) and Malik et al. (1992) Exp Hematol. September; 20(8): 1028-35 (pegylation of GM-CSF using tresyl chloride)). For example, polyethylene glycol may be covalently bound through amino acid residues via a reactive group, such as a free amino or carboxyl group. Reactive groups are those to which an activated polyethylene glycol molecule may be bound. The amino acid residues having a free amino group may include lysine residues and the N-terminal amino acid residues; those having a free carboxyl group may include aspartic acid residues, glutamic acid residues and the C-terminal amino acid residue. Sulfhydryl groups may also be used as a reactive group for attaching the polyethylene glycol molecules. For therapeutic purposes, the attachment sometimes is at an amino group, such as attachment at the N-terminus or lysine group.


Proteins can be chemically modified at the N-terminus. Using polyethylene glycol as an illustration of such a composition, one may select from a variety of polyethylene glycol molecules (by molecular weight, branching, etc.), the proportion of polyethylene glycol molecules to protein (polypeptide) molecules in the reaction mix, the type of pegylation reaction to be performed, and the method of obtaining the selected N-terminally pegylated protein. The method of obtaining the N-terminally pegylated preparation (i.e., separating this moiety from other monopegylated moieties if necessary) may be by purification of the N-terminally pegylated material from a population of pegylated protein molecules. Selective proteins chemically modified at the N-terminus may be accomplished by reductive alkylation, which exploits differential reactivity of different types of primary amino groups (lysine versus the N-terminal) available for derivatization in a particular protein. Under the appropriate reaction conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group containing polymer is achieved.


Substantially Identical NRP1, NID2. ENDO180. CDK10. FPGT. CARK, PCLO or REPS2 Nucleic Acids and Polypeptides


Nucleotide sequences and polypeptide sequences that are substantially identical to the nucleotide sequence of SEQ ID NO: 1-17 and the polypeptide sequences of SEQ ID NO: 18-27 are included herein. The term “substantially identical” as used herein refers to two or more nucleic acids or polypeptides sharing one or more identical nucleotide sequences or polypeptide sequences, respectively. Included are nucleotide sequences or polypeptide sequences that are 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more (each often within a 1%, 2%, 3% or 4% variability) identical to the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence in SEQ ID NO: 1-17 or the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide sequences of SEQ ID NO: 18-27. One test for determining whether two nucleic acids are substantially identical is to determine the percent of identical nucleotide sequences or polypeptide sequences shared between the nucleic acids or polypeptides.


Calculations of sequence identity are often performed as follows. Sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). The length of a reference sequence aligned for comparison purposes is sometimes 30% or more, 40% or more, 50% or more, often 60% or more, and more often 70%, 80%, 90%, 100% of the length of the reference sequence. The nucleotides or amino acids at corresponding nucleotide or polypeptide positions, respectively, are then compared among the two sequences. When a position in the first sequence is occupied by the same nucleotide or amino acid as the corresponding position in the second sequence, the nucleotides or amino acids are deemed to be identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, introduced for optimal alignment of the two sequences.


Comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. Percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of Meyers & Miller, CABIOS 4: 11-17 (1989), which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. Also, percent identity between two amino acid sequences can be determined using the Needleman & Wunsch, J. Mol. Biol. 48: 444-453 (1970) algorithm which has been incorporated into the GAP program in the GCG software package (available at the http address www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. Percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at http address www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. A set of parameters often used is a Blossum 62 scoring matrix with a gap open penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.


Another manner for determining if two nucleic acids are substantially identical is to assess whether a polynucleotide homologous to one nucleic acid will hybridize to the other nucleic acid under stringent conditions. As use herein, the term “stringent conditions” refers to conditions for hybridization and washing. Stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y., 6.3.1-6.3.6 (1989). Aqueous and non-aqueous methods are described in that reference and either can be used. An example of stringent hybridization conditions is hybridization in 6× sodium chloride/sodium citrate (SSC) at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 50° C. Another example of stringent hybridization conditions are hybridization in 6× sodium chloride/sodium citrate (SSC) at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 55° C. A further example of stringent hybridization conditions is hybridization in 6× sodium chloride/sodium citrate (SSC) at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 60° C. Often, stringent hybridization conditions are hybridization in 6× sodium chloride/sodium citrate (SSC) at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 65° C. More often, stringency conditions are 0.5M sodium phosphate, 7% SDS at 65° C., followed by one or more washes at 0.2×SSC, 1% SDS at 65° C.


An example of a substantially identical nucleotide sequence to SEQ ID NO: 1-17 is one that has a different nucleotide sequence and still encodes a polypeptide sequence set forth in SEQ ID NO: 18-25. Another example is a nucleotide sequence that encodes a polypeptide having a polypeptide sequence that is more than 70% identical to, sometimes more than 75%, 80%, or 85% identical to, and often more than 90% and 95% or more identical to the polypeptide sequences set forth in SEQ ID NO: 18-25.


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequences and polypeptide sequences can be used as “query sequences” to perform a search against public databases to identify other family members or related sequences, for example. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al., J. Mol. Biol. 215: 403-10 (1990). BLAST nucleotide searches can be performed with the NBLAST program, score=100, wordlength=12 to obtain nucleotide sequences homologous to NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid molecules. BLAST polypeptide searches can be performed with the XBLAST program, score=50, wordlength=3 to obtain amino acid sequences homologous to NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., Nucleic Acids Res. 25(17): 3389-3402 (1997). When utilizing BLAST and Gapped BLAST programs, default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used (see the http address www.ncbi.nlm.nih.gov).


A nucleic acid that is substantially identical to the nucleotide sequence of SEQ ID NO: 1-17 may include polymorphic sites at positions equivalent to those described herein (e.g., position 32137 in SEQ ID NO: 1) when the sequences are aligned. For example, using the alignment procedures described herein, SNPs in a sequence substantially identical to the sequence of SEQ ID NO: 1-17 can be identified at nucleotide positions that match (i.e., align) with nucleotides at SNP positions in SEQ ID NO: 1-17. Also, where a polymorphic variation is an insertion or deletion, insertion or deletion of a nucleotide sequence from a reference sequence can change the relative positions of other polymorphic sites in the nucleotide sequence.


Substantially identical NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide and polypeptide sequences include those that are naturally occurring, such as allelic variants (same locus), splice variants, homologs (different locus), and orthologs (different organism) or can be non-naturally occurring. Non-naturally occurring variants can be generated by mutagenesis techniques, including those applied to polynucleotides, cells, or organisms. The variants can contain nucleotide substitutions, deletions, inversions and insertions. Variation can occur in either or both the coding and non-coding regions. The variations can produce both conservative and non-conservative amino acid substitutions (as compared in the encoded product). Orthologs, homologs, allelic variants, and splice variants can be identified using methods known in the art. These variants normally comprise a nucleotide sequence encoding a polypeptide that is 50%, about 55% or more, often about 70-75% or more, more often about 80-85% or more, and typically about 90-95% or more identical to the amino acid sequences shown in SEQ ID NO: 18-27 or a fragment thereof. Such nucleic acid molecules can readily be identified as being able to hybridize under stringent conditions to the nucleotide sequence shown in SEQ ID NO: 1-17 or a fragment of this sequence. Nucleic acid molecules corresponding to orthologs, homologs, and allelic variants of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence can further be identified by mapping the sequence to the same chromosome or locus as the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence or variant.


Also, substantially identical NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequences may include codons that are altered with respect to the naturally occurring sequence for enhancing expression of a NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or polypeptide variant in a particular expression system. For example, the nucleic acid can be one in which one or more codons are altered, and often 10% or more or 20% or more of the codons are altered for optimized expression in bacteria (e.g., E. coli.), yeast (e.g., S. cervesiae), human (e.g., 293 cells), insect, or rodent (e.g., hamster) cells.


Methods for Identifying Subjects at Risk of Melanoma and Risk of Melanoma


Methods for determining whether a subject is at risk of melanoma are provided herein. These methods include detecting the presence or absence of one or more polymorphic variations associated with melanoma in a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence, or substantially identical sequence thereof, in a sample from a subject, where the presence of such a polymorphic variation is indicative of the subject being at risk of melanoma. These genetic tests are useful for prognosing and/or diagnosing melanoma and often are useful for determining whether an individual is at an increased, intermediate or decreased risk of developing or having melanoma.


Thus, featured herein is a method for identifying a subject at risk of melanoma, which comprises detecting in a nucleic acid sample from the subject the presence or absence of a polymorphic variation associated with melanoma at a polymorphic site in a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence. The nucleotide sequence often is selected from the group consisting of: (a) a nucleotide sequence of SEQ ID NO: 1-17; (b) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1-17; (c) a nucleotide sequence which encodes a polypeptide that is 90% or more identical to the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1-17; and (d) a fragment of a nucleotide sequence of (a), (b), or (c), where the fragment often comprises a polymorphic site; whereby the presence of the polymorphic variation is indicative of the subject being at risk of melanoma. A polymorphic variation assayed in the genetic test often is located in an intron, sometimes in a region surrounding the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 open reading frame (e.g., within 50 kilobases (kb), 40 kb, 30 kb, 20 kb, 15, kb, 10 kb, 5 kb, 4 kb, 3 kb, 2 kb, or 1 kb of the open reading frame initiation site or termination site), and sometimes in an exon. Sometimes the polymorphic variation is not located in an exon (e.g., it sometimes is located in an intron or region upstream or downstream of a terminal intron or exon). As used herein, “SEQ ID NO: 1-17” refers to individual sequences in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17, each sequence being separately applicable to embodiments described herein.


Results from such genetic tests may be combined with other test results to diagnose melanoma. For example, genetic test results may be gathered, a patient sample may be ordered based on a determined predisposition to melanoma (e.g., a skin biopsy), the patient sample is analyzed, and the results of the analysis may be utilized to diagnose melanoma. Also, melanoma diagnostic tests are generated by stratifying populations into subpopulations having different progressions of melanoma and detecting polymorphic variations associated with different progressions of the melanoma, as described in further detail hereafter. In another embodiment, genetic test results are gathered, a patient's risk factors for developing melanoma are analyzed (e.g., exposure to sun and skin pigmentation), and a patient sample may be ordered based on a determined risk of melanoma.


Risk of melanoma sometimes is expressed as a probability, such as an odds ratio, percentage, or risk factor. The risk assessment is based upon the presence or absence of one or more polymorphic variants described herein, and also may be based in part upon phenotypic traits of the individual being tested. Methods for calculating risks based upon patient data are well known (see, e.g., Agresti, Categorical Data Analysis, 2nd Ed. 2002. Wiley). Allelotyping and genotyping analyses may be carried out in populations other than those exemplified herein to enhance the predictive power of the prognostic method. These further analyses are executed in view of the exemplified procedures described herein, and may be based upon the same polymorphic variations or additional polymorphic variations.


The nucleic acid sample typically is isolated from a biological sample obtained from a subject. For example, nucleic acid can be isolated from blood, saliva, sputum, urine, cell scrapings, and biopsy tissue. The nucleic acid sample can be isolated from a biological sample using standard techniques, such as the technique described in Example 2. As used herein, the term “subject” refers primarily to humans but also refers to other mammals such as dogs, cats, and ungulates (e.g., cattle, sheep, and swine). Subjects also include avians (e.g., chickens and turkeys), reptiles, and fish (e.g., salmon), as embodiments described herein can be adapted to nucleic acid samples isolated from any of these organisms. The nucleic acid sample may be isolated from the subject and then directly utilized in a method for determining the presence of a polymorphic variant, or alternatively, the sample may be isolated and then stored (e.g., frozen) for a period of time before being subjected to analysis.


The presence or absence of a polymorphic variant is determined using one or both chromosomal complements represented in the nucleic acid sample. Determining the presence or absence of a polymorphic variant in both chromosomal complements represented in a nucleic acid sample from a subject having a copy of each chromosome is useful for determining the zygosity of an individual for the polymorphic variant (i.e., whether the individual is homozygous or heterozygous for the polymorphic variant). Any oligonucleotide-based diagnostic may be utilized to determine whether a sample includes the presence or absence of a polymorphic variant in a sample. For example, primer extension methods, ligase sequence determination methods (e.g., U.S. Pat. Nos. 5,679,524 and 5,952,174, and WO 01/27326), mismatch sequence determination methods (e.g., U.S. Pat. Nos. 5,851,770; 5,958,692; 6,110,684; and 6,183,958), microarray sequence determination methods, restriction fragment length polymorphism (RFLP), single strand conformation polymorphism detection (SSCP) (e.g., U.S. Pat. Nos. 5,891,625 and 6,013,499), PCR-based assays (e.g., TAQMAN® PCR System (Applied Biosystems)), and nucleotide sequencing methods may be used.


Oligonucleotide extension methods typically involve providing a pair of oligonucleotide primers in a polymerase chain reaction (PCR) or in other nucleic acid amplification methods for the purpose of amplifying a region from the nucleic acid sample that comprises the polymorphic variation. One oligonucleotide primer is complementary to a region 3′ of the polymorphism and the other is complementary to a region 5′ of the polymorphism. A PCR primer pair may be used in methods disclosed in U.S. Pat. Nos. 4,683,195; 4,683,202, 4,965,188; 5,656,493; 5,998,143; 6,140,054; WO 01/27327; and WO 01/27329 for example. PCR primer pairs may also be used in any commercially available machines that perform PCR, such as any of the GENEAMP® Systems available from Applied Biosystems. Also, those of ordinary skill in the art will be able to design oligonucleotide primers based upon a nucleotide sequence of SEQ ID NO: 1-17 without undue experimentation using knowledge readily available in the art.


Also provided are extension oligonucleotides that hybridize to the amplified fragment adjacent to the polymorphic variation. As used herein, the term “adjacent” refers to the 3′ end of the extension oligonucleotide being sometimes 1 nucleotide from the 5′ end of the polymorphic site, often 2 or 3, and at times 4, 5, 6, 7, 8, 9, or 10 nucleotides from the 5′ end of the polymorphic site, in the nucleic acid when the extension oligonucleotide is hybridized to the nucleic acid. The extension oligonucleotide then is extended by one or more nucleotides, often 2 or 3 nucleotides, and the number and/or type of nucleotides that are added to the extension oligonucleotide determine whether the polymorphic variant is present. Oligonucleotide extension methods are disclosed, for example, in U.S. Pat. Nos. 4,656,127; 4,851,331; 5,679,524; 5,834,189; 5,876,934; 5,908,755; 5,912,118; 5,976,802; 5,981,186; 6,004,744; 6,013,431; 6,017,702; 6,046,005; 6,087,095; 6,210,891; and WO 01/20039. Oligonucleotide extension methods using mass spectrometry are described, for example, in U.S. Pat. Nos. 5,547,835; 5,605,798; 5,691,141; 5,849,542; 5,869,242; 5,928,906; 6,043,031; and 6,194,144, and a method often utilized is described herein in Example 2.


A microarray can be utilized for determining whether a polymorphic variant is present or absent in a nucleic acid sample. A microarray sometimes includes an oligonucleotides described herein, and methods for making and using oligonucleotide microarrays suitable for prognostic use are disclosed in U.S. Pat. Nos. 5,492,806; 5,525,464; 5,589,330; 5,695,940; 5,849,483; 6,018,041; 6,045,996; 6,136,541; 6,142,681; 6,156,501; 6,197,506; 6,223,127; 6,225,625; 6,229,911; 6,239,273; WO 00/52625; WO 01/25485; and WO 01/29259. The microarray typically comprises a solid support and the oligonucleotides sometimes are linked to the solid support by covalent or non-covalent interactions. The oligonucleotides sometimes are linked to the solid support directly or by a spacer molecule. A microarray sometimes comprise one or more oligonucleotides complementary to a portion of SEQ ID NO: 1-17, or complementary to a variant described herein.


A kit may also be utilized for determining whether a polymorphic variant is present or absent in a nucleic acid sample. A kit often comprises one or more pairs of oligonucleotide primers useful for amplifying a fragment of SEQ ID NO: 1-17 or a substantially identical sequence thereof, where the fragment includes a polymorphic site. The kit sometimes comprises a polymerizing agent, for example, a thermostable nucleic acid polymerase such as one disclosed in U.S. Pat. No. 4,889,818 or 6,077,664. Also, the kit often comprises an elongation oligonucleotide that hybridizes to a NRP1, NID2, ENDO180, CDK180, FPGT, CARK, PCLO or REPS2 nucleic acid in a nucleic acid sample adjacent to the polymorphic site. Where the kit includes an elongation oligonucleotide, it also often comprises chain elongating nucleotides, such as dATP, dTTP, dGTP, dCTP, and dITP, including analogs of dATP, dTTP, dGTP, dCTP and dITP, provided that such analogs are substrates for a thermostable nucleic acid polymerase and can be incorporated into a nucleic acid chain elongated from the extension oligonucleotide. Along with chain elongating nucleotides would be one or more chain terminating nucleotides such as ddATP, ddTTP, ddGTP, ddCTP, and the like. In an embodiment, the kit comprises one or more oligonucleotide primer pairs, a polymerizing agent, chain elongating nucleotides, at least one elongation oligonucleotide, and one or more chain terminating nucleotides. Kits optionally include buffers, vials, microtitre plates, and instructions for use. NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed hits may be utilized to prognose or diagnose melanoma for a significant fraction of melanoma occurrences, such as in 50% or more melanoma occurrences, or sometimes 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more melanoma occurrences.


Using a polymorphism detection technology (e.g., a technique described above or below in Example 2), mutations and polymorphisms in or around the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 locus may be detected in melanocytic lesions, which include nevi, radial growth phase (RGP) melanomas, vertical growth phase (VGP) melanomas, and melanoma metastases. The mutations can be detected within 50 kilobases (kb), 40 kb, 30 kb, 20 kb, 15, kb, 10 kb, 5 kb, 4 kb, 3 kb, 2 kb, or 1 kb from the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 open reading frame initiation or termination site. Therefore, provided herein are methods for genotyping NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mutations in melanocytic lesions and metastases (e.g., described in Example 2). Mutations in or around the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 loci present in later stage melanomas, such as VGP melanomas and melanoma metastases, are indicative of melanomas particularly likely to continue to progress and/or metastasize (e.g., from RGP to VGP melanoma or melanoma metastases), i.e., aggressive melanomas. Thus, provided herein are methods for identifying subjects at risk of a progressive or aggressive melanoma by determining the presence or absence of one or more NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mutations in the DNA sample of a subject that exist in melanocytic lesions and/or metastases. Identifying the presence of one or more of these mutations is useful for identifying subjects in need of aggressive treatments of melanoma, and once identified using such methods, a subject often is given information concerning preventions and treatments of the disease, and sometimes is treated with an aggressive melanoma treatment method (e.g., surgery or administration of drugs), as described in more detail hereafter.


Determining the presence of a polymorphic variant, or a combination of two or more polymorphic variants, in a nucleic acid set forth in SEQ ID NO: 1-17, or SEQ ID NO: 1-7, of the sample often is indicative of a predisposition to melanoma. In certain embodiments, nucleic acid variants of other loci, such as the BRAF locus described in U.S. application Ser. No. 10/661,966 filed Sep. 11, 2003 for example, are detected in combination with one or more nucleic acid variants in the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 loci.


The following variants are associated with an increased risk of melanoma: a thymine at the chromosome position indicated by rs1360457, an adenine at the chromosome position indicated by rs753050, a guanine at the chromosome position indicated by rs729647, a thymine at the chromosome position indicated by rs3742536, a guanine at position 32137 in SEQ ID NO: 1, a thymine at position 12008 in SEQ ID NO: 1, a cytosine at position 32720 in SEQ ID NO: 1, an adenine at position 43721 in SEQ ID NO: 1, a guanine at position 44339 in SEQ ID NO: 1, a thymine at position 45640 in SEQ ID NO: 1, an adenine at position 48768 in SEQ ID NO: 1, a guanine at position 74247 in SEQ ID NO: 1, a thymine at position 75828 in SEQ ID NO: 1, an adenine at position 76381 in SEQ ID NO: 1, and a cytosine at position 84909. An individual identified as being at risk of melanoma may be heterozygous or homozygous with respect to the allele associated with melanoma. A subject homozygous for an allele associated with an increased risk of melanoma (e.g., a thymine at position 45640 in SEQ ID NO: 1) is at a comparatively high risk of melanoma, a subject heterozygous for an allele associated with an increased risk of melanoma is at a comparatively intermediate risk of melanoma, and a subject homozygous for an allele associated with a decreased risk of melanoma (e.g., a cytosine at position 45640 in SEQ ID NO: 1, see Examples section below) is at a comparatively lower risk of melanoma. A genotype may be assessed for a complementary strand, such that the complementary nucleotide at a particular position is detected (e.g., if a thymine or cytosine is detected at position 45640 in SEQ ID NO: 1, the complementary strand would yield an adenine or guanine, respectively, where the adenine is associated with increased risk of melanoma).


Also featured are methods for determining risk of melanoma and/or identifying a subject at risk of melanoma by contacting a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or protein from a subject with an antibody that specifically binds to an epitope associated with increased risk of melanoma in the polypeptide. In an embodiment, the antibody specifically binds to an epitope that comprises a serine at amino acid position 656 in a NID2 polypeptide.


Applications of Genomic Information to Pharmacogenomics


Pharmacogenomics is a discipline that involves tailoring a treatment for a subject according to the subject's genotype as a particular treatment regimen may exert a differential effect depending upon the subject's genotype. Based upon the outcome of a prognostic test described herein, a clinician or physician may target pertinent information and preventative or therapeutic treatments to a subject who would be benefited by the information or treatment and avoid directing such information and treatments to a subject who would not be benefited (e.g., the treatment has no therapeutic effect and/or the subject experiences adverse side effects).


For example, where a candidate therapeutic exhibits a significant interaction with a major allele and a comparatively weak interaction with a minor allele (e.g., an order of magnitude or greater difference in the interaction), such a therapeutic typically would not be administered to a subject genotyped as being homozygous for the minor allele, and sometimes not administered to a subject genotyped as being heterozygous for the minor allele. In another example, where a candidate therapeutic is not significantly toxic when administered to subjects who are homozygous for a major allele but is comparatively toxic when administered to subjects heterozygous or homozygous for a minor allele, the candidate therapeutic is not typically administered to subjects who are genotyped as being heterozygous or homozygous with respect to the minor allele.


The prognostic methods described herein are applicable to general pharmacogenomic approaches towards addressing melanoma. For example, a nucleic acid sample from an individual may be subjected to a prognostic test described herein. Where one or more polymorphic variations associated with increased risk of melanoma are identified in a subject, one or more melanoma treatments or prophylactic regimens may be prescribed to that subject. Subjects genotyped as having one or more of the alleles described herein that are associated with increased risk of melanoma often are prescribed a prophylactic regimen designed to minimize the occurrence of melanoma. An example of a prophylactic regimen often prescribed is directed towards minimizing ultraviolet (UV) light exposure. Such a regimen may include, for example, prescription of a lotion applied to the skin that minimizes UV penetration and/or counseling individuals of other practices for reducing UV exposure, such as by wearing protective clothing and minimizing sun exposure.


In certain embodiments, a treatment regimen is specifically prescribed and/or administered to individuals who will most benefit from it based upon their risk of developing melanoma assessed by the prognostic methods described herein. Thus, provided are methods for identifying a subject predisposed to melanoma and then prescribing a therapeutic or preventative regimen to individuals identified as having a predisposition. Thus, certain embodiments are directed to a method for reducing melanoma in a subject, which comprises: detecting the presence or absence of a polymorphic variant associated with melanoma in a nucleotide sequence of a nucleic acid sample from a subject, where the nucleotide sequence comprises a polynucleotide sequence selected from the group consisting of: (a) a nucleotide sequence set forth in SEQ ID NO: 1-17; (b) a nucleotide sequence which encodes a polypeptide consisting of an amino acid sequence encoded by a nucleotide sequence of SEQ ID NO: 1-17; (c) a nucleotide sequence which encodes a polypeptide that is 90% or more identical to an amino acid sequence encoded by a nucleotide sequence of SEQ ID NO: 1-17 or a nucleotide sequence about 90% or more identical to the nucleotide sequence in SEQ ID NO: 1-17; and (d) a fragment of a polynucleotide sequence of (a), (b), or (c); and prescribing or administering a treatment regimen to a subject from whom the sample originated where the presence of a polymorphic variation associated with melanoma is detected in the nucleotide sequence. In these methods, predisposition results may be utilized in combination with other test results to diagnose melanoma.


The treatment sometimes is preventative (e.g., is prescribed or administered to reduce the probability that a melanoma associated condition arises or progresses), sometimes is therapeutic, and sometimes delays, alleviates or halts the progression of a melanoma associated condition. Any known preventative or therapeutic treatment for alleviating or preventing the occurrence of a melanoma associated disorder is prescribed and/or administered. For example, the treatment sometimes is or includes a drug that reduces melanoma, including, for example, cisplatin, carmustine (BCNU), vinblastine, vincristine, and bleomycin, and/or a molecule that interacts with a nucleic acid or polypeptide described hereafter. In another example, the melanoma treatment is surgery. Surgery to remove (excise) a melanoma is the standard treatment for this disease. It is necessary to remove not only the tumor but also some normal tissue around it in order to minimize the chance that any cancer will be left in the area. It is common for lymph nodes near the tumor to be removed during surgery because cancer can spread through the lymphatic system. Surgery is generally not effective in controlling melanoma that is known to have spread to other parts of the body. In such cases, doctors may use other methods of treatment, such as chemotherapy, biological therapy, radiation therapy, or a combination of these methods.


As therapeutic approaches for melanoma continue to evolve and improve, the goal of treatments for melanoma related disorders is to intervene even before clinical signs (e.g., identification of irregular nevi based on A—asymmetry, B—border irregularity, C—color variation, D—diameter of >6 mm as described by Friedman R J, et al. in CA Cancer J. Clin. 1985 May-June; 35(3): 130-51) first manifest. Thus, genetic markers associated with susceptibility to melanoma prove useful for early diagnosis, prevention and treatment of melanoma.


As melanoma preventative and treatment information can be specifically targeted to subjects in need thereof (e.g., those at risk of developing melanoma or those that have early signs of melanoma), provided herein is a method for preventing or reducing the risk of developing melanoma in a subject, which comprises: (a) detecting the presence or absence of a polymorphic variation associated with melanoma at a polymorphic site in a nucleotide sequence in a nucleic acid sample from a subject; (b) identifying a subject with a predisposition to melanoma, whereby the presence of the polymorphic variation is indicative of a predisposition to melanoma in the subject; and (c) if such a predisposition is identified, providing the subject with information about methods or products to prevent or reduce melanoma or to delay the onset of melanoma. Also provided is a method of targeting information or advertising to a subpopulation of a human population based on the subpopulation being genetically predisposed to a disease or condition, which comprises: (a) detecting the presence or absence of a polymorphic variation associated with melanoma at a polymorphic site in a nucleotide sequence in a nucleic acid sample from a subject; (b) identifying the subpopulation of subjects in which the polymorphic variation is associated with melanoma; and (c) providing information only to the subpopulation of subjects about a particular product which may be obtained and consumed or applied by the subject to help prevent or delay onset of the disease or condition.


Pharmacogenomics methods also may be used to analyze and predict a response to a melanoma treatment or a drug. For example, if pharmacogenomics analysis indicates a likelihood that an individual will respond positively to a melanoma treatment with a particular drug, the drug may be administered to the individual. Conversely, if the analysis indicates that an individual is likely to respond negatively to treatment with a particular drug, an alternative course of treatment may be prescribed. A negative response may be defined as either the absence of an efficacious response or the presence of toxic side effects. The response to a therapeutic treatment can be predicted in a background study in which subjects in any of the following populations are genotyped: a population that responds favorably to a treatment regimen, a population that does not respond significantly to a treatment regimen, and a population that responds adversely to a treatment regiment (e.g., exhibits one or more side effects). These populations are provided as examples and other populations and subpopulations may be analyzed. Based upon the results of these analyses, a subject is genotyped to predict whether he or she will respond favorably to a treatment regimen, not respond significantly to a treatment regimen, or respond adversely to a treatment regimen.


The tests described herein also are applicable to clinical drug trials. One or more polymorphic variants indicative of response to an agent for treating melanoma or to side effects to an agent for treating melanoma may be identified using the methods described herein. Thereafter, potential participants in clinical trials of such an agent may be screened to identify those individuals most likely to respond favorably to the drug and exclude those likely to experience side effects. In that way, the effectiveness of drug treatment may be measured in individuals who respond positively to the drug, without lowering the measurement as a result of the inclusion of individuals who are unlikely to respond positively in the study and without risking undesirable safety problems.


Thus, another embodiment is a method of selecting an individual for inclusion in a clinical trial of a treatment or drug comprising the steps of: (a) obtaining a nucleic acid sample from an individual; (b) determining the identity of a polymorphic variation which is associated with a positive response to the treatment or the drug, or at least one polymorphic variation which is associated with a negative response to the treatment or the drug in the nucleic acid sample, and (c) including the individual in the clinical trial if the nucleic acid sample contains said polymorphic variation associated with a positive response to the treatment or the drug or if the nucleic acid sample lacks said polymorphic variation associated with a negative response to the treatment or the drug. In addition, the methods for selecting an individual for inclusion in a clinical trial of a treatment or drug encompass methods with any further limitation described in this disclosure, or those following, specified alone or in any combination. The polymorphic variation may be in a sequence selected individually or in any combination from the group consisting of (i) a polynucleotide sequence set forth in SEQ ID NO: 1-17; (ii) a polynucleotide sequence that is 90% or more identical to an amino acid sequence encoded by a nucleotide sequence of SEQ ID NO: 1-17; (iii) a polynucleotide sequence that encodes a polypeptide having an amino acid sequence identical to or 90% or more identical to an amino acid sequence encoded by a nucleotide sequence of SEQ ID NO: 1-17; and (iv) a fragment of a polynucleotide sequence of (i), (ii), or (iii) comprising the polymorphic site. The including step (c) optionally comprises administering the drug or the treatment to the individual if the nucleic acid sample contains the polymorphic variation associated with a positive response to the treatment or the drug and the nucleic acid sample lacks said biallelic marker associated with a negative response to the treatment or the drug.


Also provided herein is a method of partnering between a diagnostic/prognostic testing provider and a provider of a consumable product, which comprises: (a) the diagnostic/prognostic testing provider detects the presence or absence of a polymorphic variation associated with melanoma at a polymorphic site in a nucleotide sequence in a nucleic acid sample from a subject; (b) the diagnostic/prognostic testing provider identifies the subpopulation of subjects in which the polymorphic variation is associated with melanoma; (c) the diagnostic/prognostic testing provider forwards information to the subpopulation of subjects about a particular product which may be obtained and consumed or applied by the subject to help prevent or delay onset of the disease or condition; and (d) the provider of a consumable product forwards to the diagnostic test provider a fee every time the diagnostic/prognostic test provider forwards information to the subject as set forth in step (c) above.


Compositions Comprising a NRP1. NID2. ENDO180. CDK10. FPGT. CARK. PCLO or REPS2 Directed Molecule


Featured herein is a composition comprising a melanoma cell and one or more NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed molecules. NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed molecules include, but are not limited to, a compound that binds to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or polypeptide; an RNAi or siRNA molecule having a strand complementary to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 DNA sequence; an antisense nucleic acid complementary to an RNA encoded by a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 DNA sequence; a nucleic acid complementary to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 RNA, cDNA or gDNA nucleotide sequence, sometimes linked to a solid support or an array; a ribozyme that hybridizes to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence; an NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, protein or fragment thereof, or a nucleic acid that encodes the foregoing; a nucleic acid aptamer that specifically binds a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, protein, nucleic acid or variant thereof; and an antibody or fragment thereof that specifically binds to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, protein, nucleic acid or variant thereof. Compositions comprising an antibody often include an adjuvant known in the art. The melanoma cell may be in a group of melanoma cells and/or other types of cells cultured in vitro or in a tissue having melanoma cells (e.g., a melanocytic lesion) maintained in vitro or present in an animal in vivo (e.g., a rat, mouse, ape or human). In certain embodiments, a composition comprises a component from a melanoma cell or from a subject having a melanoma cell instead of the melanoma cell or in addition to the melanoma cell, where the component sometimes is a nucleic acid molecule (e.g., genomic DNA), a protein mixture or isolated protein, for example. The aforementioned compositions have utility in diagnostic, prognostic and pharmacogenomic methods described previously and in melanoma therapeutics described hereafter. Certain NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed molecules are described in greater detail below.


Compounds


Compounds can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; peptoid libraries (libraries of molecules having the functionalities of peptides, but with a novel, non-peptide backbone which are resistant to enzymatic degradation but which nevertheless remain bioactive (see, e.g., Zuckermann et al., J. Med. Chem. 37: 2678-85 (1994)); spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; “one-bead one-compound” library methods; and synthetic library methods using affinity chromatography selection. Biological library and peptoid library approaches are typically limited to peptide libraries, while the other approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds (Lam, Anticancer Drug Des. 12: 145, (1997)). Examples of methods for synthesizing molecular libraries are described, for example, in DeWitt et al., Proc. Natl. Acad. Sci. USA. 90: 6909 (1993); Erb et al., Proc. Natl. Acad. Sci. USA 91: 11422 (1994); Zuckermann et al., J. Med. Chem. 37: 2678 (1994); Cho et al., Science 261: 1303 (1993); Carrell et al., Angew. Chem. Int. Ed. Engl. 33: 2059 (1994); Carell et al., Angew. Chem. Int. Ed. Engl. 33: 2061 (1994); and in Gallop et al., J. Med. Chem. 37: 1233 (1994).


Libraries of compounds may be presented in solution (e.g., Houghten, Biotechniques 13: 412421 (1992)), or on beads (Lam, Nature 354: 82-84 (1991)), chips (Fodor, Nature 364: 555-556 (1993)), bacteria or spores (Ladner, U.S. Pat. No. 5,223,409), plasmids (Cull et al., Proc. Natl. Acad. Sci. USA 89: 1865-1869 (1992)) or on phage (Scott and Smith, Science 249: 386-390 (1990); Devlin, Science 249: 404-406 (1990); Cwirla et al., Proc. Natl. Acad. Sci. 87: 6378-6382 (1990); Felici, J. Mol. Biol. 222: 301-310 (1991); Ladner supra.).


A compound may alter expression or activity of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides and may be a small molecule. Small molecules include, but are not limited to, peptides, peptidomimetics (e.g., peptoids), amino acids, amino acid analogs, polynucleotides, polynucleotide analogs, nucleotides, nucleotide analogs, organic or inorganic compounds (i.e., including heteroorganic and organometallic compounds) having a molecular weight less than about 10,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 5,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 1,000 grams per mole, organic or inorganic compounds having a molecular weight less than about 500 grams per mole, and salts, esters, and other pharmaceutically acceptable forms of such compounds.


Antisense Nucleic Acid Molecules, Ribozymes, RNAi, siRNA and Modified NRP1, NID2. ENDO180. CDK10. FPGT, CARK, PCLO or REPS2 Nucleic Acid Molecules


An “antisense” nucleic acid refers to a nucleotide sequence complementary to a “sense” nucleic acid encoding a polypeptide, e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence. The antisense nucleic acid can be complementary to an entire NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 coding strand, or to only a portion thereof (e.g., coding regions of human NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 in SEQ ID NO: 1-17). In another embodiment, the antisense nucleic acid molecule is antisense to a “noncoding region” of the coding strand of a nucleotide sequence encoding NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 (e.g., 5′ and 3′ untranslated regions).


An antisense nucleic acid can be designed such that it is complementary to the entire coding region of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA, and often the antisense nucleic acid is an oligonucleotide antisense to only a portion of a coding or noncoding region of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA, e.g., between the −10 and +10 regions of the target gene nucleotide sequence of interest. An antisense oligonucleotide can be, for example, about 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, or more nucleotides in length. The antisense nucleic acids, which include the ribozymes described hereafter, can be designed to target NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid variants. Among the variants, minor alleles and major alleles can be targeted, and those associated with a higher risk of melanoma are often designed, tested, and administered to subjects.


An antisense nucleic acid can be constructed using chemical synthesis and enzymatic ligation reactions using standard procedures. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used. Antisense nucleic acid also can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection).


When utilized as therapeutics, antisense nucleic acids typically are administered to a subject (e.g., by direct injection at a tissue site) or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide and thereby inhibit expression of the polypeptide, for example, by inhibiting transcription and/or translation. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then are administered systemically. For systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface, for example, by linking antisense nucleic acid molecules to peptides or antibodies which bind to cell surface receptors or antigens. Antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. Sufficient intracellular concentrations of antisense molecules are achieved by incorporating a strong promoter, such as a pol II or pol III promoter, in the vector construct.


Antisense nucleic acid molecules sometimes are α-anomeric nucleic acid molecules. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other (Gaultier et al., Nucleic Acids. Res. 15: 6625-6641 (1987)). Antisense nucleic acid molecules can also comprise a 2′—O— methylribonucleotide (Inoue et al., Nucleic Acids Res. 15: 6131-6148 (1987)) or a chimeric RNA-DNA analogue (Inoue et al., FEBS Lett. 215: 327-330 (1987)). Antisense nucleic acids sometimes are composed of DNA or PNA or any other nucleic acid derivatives described previously.


In another embodiment, an antisense nucleic acid is a ribozyme. A ribozyme having specificity for a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 encoding nucleic acid can include one or more sequences complementary to the nucleotide sequence of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 DNA sequence disclosed herein (e.g., SEQ ID NO: 1-17), and a sequence having a known catalytic region responsible for mRNA cleavage (see e.g., U.S. Pat. No. 5,093,246 or Haselhoff and Gerlach, Nature 334: 585-591 (1988)). For example, a derivative of a Tetrahymena L-19 IVS RNA is sometimes utilized in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 encoding mRNA (see e.g., Cech et al. U.S. Pat. No. 4,987,071; and Cech et al. U.S. Pat. No. 5,116,742). Also, NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules (see e.g., Bartel & Szostak, Science 261: 1411-1418 (1993)).


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed molecules include in certain embodiments nucleic acids that can form triple helix structures with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence, especially one that includes a regulatory region that controls NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression. NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 (e.g., NRP1, NID2, ENDO80, CDK10, FPGT, CARK, PCLO or REPS2 promoter and/or enhancers) to form triple helical structures that prevent transcription of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 gene in target cells (see e.g., Helene, Anticancer Drug Des. 6(6): 569-84 (1991); Helene et al., Ann. N.Y. Acad. Sci. 660: 27-36 (1992); and Maher, Bioassays 14(12): 807-15 (1992). Potential sequences that can be targeted for triple helix formation can be increased by creating a so-called “switchback” nucleic acid molecule. Switchback molecules are synthesized in an alternating 5′-3′,3′-5′ manner, such that they base pair with first one strand of a duplex and then the other, eliminating the necessity for a sizeable stretch of either purines or pyrimidines to be present on one strand of a duplex.


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 directed molecules include RNAi and siRNA nucleic acids. Gene expression may be inhibited by the introduction of double-stranded RNA (dsRNA), which induces potent and specific gene silencing, a phenomenon called RNA interference or RNAi. See, e.g., Fire et al., U.S. Pat. No. 6,506,559; Tuschl et al. PCT International Publication No. WO 01/75164; Kay et al. PCT International Publication No. WO 03/010180A1; or Bosher J M, Labouesse, Nat Cell Biol 2000 February; 2(2): E31-6. This process has been improved by decreasing the size of the double-stranded RNA to 20-24 base pairs (to create small-interfering RNAs or siRNAs) that “switched off” genes in mammalian cells without initiating an acute phase response, i.e., a host defense mechanism that often results in cell death (see, e.g., Caplen et al. Proc Natl Acad Sci USA. 2001 Aug. 14; 98(17): 9742-7 and Elbashir et al. Methods 2002 February; 26(2): 199-213). There is increasing evidence of post-transcriptional gene silencing by RNA interference (RNAi) for inhibiting targeted expression in mammalian cells at the mRNA level, in human cells. There is additional evidence of effective methods for inhibiting the proliferation and migration of tumor cells in human patients, and for inhibiting metastatic cancer development (see, e.g., U.S. patent application No. U.S.2001000993183; Caplen et al. Proc Natl Acad Sci USA; and Abderrahmani et al. Mol Cell Biol 2001 November 21(21): 7256-67).


An “siRNA” or “RNAi” refers to a nucleic acid that forms a double stranded RNA and has the ability to reduce or inhibit expression of a gene or target gene when the siRNA is delivered to or expressed in the same cell as the gene or target gene. “siRNA” refers to short double-stranded RNA formed by the complementary strands. Complementary portions of the siRNA that hybridize to form the double stranded molecule often have substantial or complete identity to the target molecule sequence. In one embodiment, an siRNA refers to a nucleic acid that has substantial or complete identity to a target gene and forms a double stranded siRNA, such as a nucleotide sequence set forth in SEQ ID NO: 1-17, for example.


When designing the siRNA molecules, the targeted region often is selected from a given DNA sequence beginning 50 to 100 nucleotides downstream of the start codon. See, e.g., Elbashir et al,. Methods 26: 199-213 (2002). Initially, 5′ or 3′ UTRs and regions nearby the start codon were avoided assuming that UTR-binding proteins and/or translation initiation complexes may interfere with binding of the siRNP or RISC endonuclease complex. Sometimes regions of the target 23 nucleotides in length conforming to the sequence motif AA(N19)TT (N, an nucleotide), and regions with approximately 30% to 70% G/C-content (often about 50% G/C-content) often are selected. If no suitable sequences are found, the search often is extended using the motif NA(N21). The sequence of the sense siRNA sometimes corresponds to (N19) TT or N21 (position 3 to 23 of the 23-nt motif), respectively. In the latter case, the 3′ end of the sense siRNA often is converted to TT. The rationale for this sequence conversion is to generate a symmetric duplex with respect to the sequence composition of the sense and antisense 3′ overhangs. The antisense siRNA is synthesized as the complement to position 1 to 21 of the 23-nt motif. Because position 1 of the 23-nt motif is not recognized sequence-specifically by the antisense siRNA, the 3′-most nucleotide residue of the antisense siRNA can be chosen deliberately. However, the penultimate nucleotide of the antisense siRNA (complementary to position 2 of the 23-nt motif) often is complementary to the targeted sequence. For simplifying chemical synthesis, TT often is utilized. siRNAs corresponding to the target motif NAR(N 17)YNN, where R is purine (A,G) and Y is pyrimidine (C,U), often are selected. Respective 21 nucleotide sense and antisense siRNAs often begin with a purine nucleotide and can also be expressed from pol III expression vectors without a change in targeting site. Expression of RNAs from pol III promoters often is efficient when the first transcribed nucleotide is a purine.


The sequence of the siRNA can correspond to the full length target gene, or a subsequence thereof. Often, the siRNA is about 15 to about 50 nucleotides in length (e.g., each complementary sequence of the double stranded siRNA is 15-50 nucleotides in length, and the double stranded siRNA is about 15-50 base pairs in length, sometimes about 20-30 nucleotides in length or about 20-25 nucleotides in length, e.g., 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. The siRNA sometimes is about 21 nucleotides in length. Methods of using siRNA are well known in the art, and specific siRNA molecules may be purchased from a number of companies including Dharmacon Research, Inc.


Antisense, ribozyme, RNAi and siRNA nucleic acids can be altered to form modified NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid molecules. The nucleic acids can be altered at base moieties, sugar moieties or phosphate backbone moieties to improve stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of nucleic acid molecules can be modified to generate peptide nucleic acids (see Hyrup et al., Bioorganic & Medicinal Chemistry 4 (1): 5-23 (1996)). As used herein, the terms “peptide nucleic acid” or “PNA” refers to a nucleic acid mimic such as a DNA mimic, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of a PNA can allow for specific hybridization to DNA and RNA under conditions of low ionic strength. Synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described, for example, in Hyrup et al., (1996) supra and Perry-O'Keefe et al., Proc. Natl. Acad. Sci. 93: 14670-675 (1996).


PNAs of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids can be used in prognostic, diagnostic, and therapeutic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, for example, inducing transcription or translation arrest or inhibiting replication. PNAs of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid molecules can also be used in the analysis of single base pair mutations in a gene, (e.g., by PNA-directed PCR clamping); as “artificial restriction enzymes” when used in combination with other enzymes, (e.g., S1 nucleases (Hyrup (1996) supra)); or as probes or primers for DNA sequencing or hybridization (Hyrup et al., (1996) supra; Perry-O'Keefe supra).


In other embodiments, oligonucleotides may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across cell membranes (see e.g., Letsinger et al., Proc. Natl. Acad. Sci. USA 86: 6553-6556 (1989); Lemaitre et al., Proc. Natl. Acad. Sci. USA 84: 648-652 (1987); PCT Publication No. WO88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO89/10134). In addition, oligonucleotides can be modified with hybridization-triggered cleavage agents (See, e.g., Krol et al., Bio-Techniques 6: 958-976 (1988)) or intercalating agents. (See, e.g., Zon, Pharm. Res. 5: 539-549 (1988)). To this end, the oligonucleotide may be conjugated to another molecule, (e.g., a peptide, hybridization triggered cross-linking agent, transport agent, or hybridization-triggered cleavage agent).


Also included herein are molecular beacon oligonucleotide primer and probe molecules having one or more regions complementary to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, two complementary regions one having a fluorophore and one a quencher such that the molecular beacon is useful for quantifying the presence of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid in a sample. Molecular beacon nucleic acids are described, for example, in Lizardi et al., U.S. Pat. No. 5,854,033; Nazarenko et al., U.S. Pat. No. 5,866,336, and Livak et al., U.S. Pat. No. 5,876,930.


Anti-NRP1. NID2. ENDO180. CDK10, FPGT. CARK. PCLO or REPS2 Antibodies


The term “antibody” as used herein refers to an immunoglobulin molecule or immunologically active portion thereof, i.e., an antigen-binding portion. Examples of immunologically active portions of immunoglobulin molecules include F(ab) and F(ab′)2 fragments which can be generated by treating the antibody with an enzyme such as pepsin. An antibody sometimes is a polyclonal, monoclonal, recombinant (e.g., a chimeric or humanized), fully human, non-human (e.g., murine), or a single chain antibody. An antibody may have effector function and can fix complement, and is sometimes coupled to a toxin or imaging agent.


A full-length NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or antigenic peptide fragment can be used as an immunogen or can be used to identify anti-NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibodies made with other immunogens, e.g., cells, membrane preparations, and the like. An antigenic peptide of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 often includes at least 8 amino acid residues of the amino acid sequences set forth in SEQ ID NO: 18-27 and encompasses an epitope of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2. Antigenic peptides sometimes include 10 or more amino acids, 15 or more amino acids, 20 or more amino acids, or 30 or more amino acids. Hydrophilic and hydrophobic fragments of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides sometimes are used as immunogens.


Epitopes encompassed by the antigenic peptide are regions of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 located on the surface of the polypeptide (e.g., hydrophilic regions) as well as regions with high antigenicity. For example, an Emini surface probability analysis of the human NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide sequence can be used to indicate the regions that have a particularly high probability of being localized to the surface of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide and are thus likely to constitute surface residues useful for targeting antibody production. The antibody may bind an epitope on any domain or region on NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides described herein.


Also, chimeric, humanized, and completely human antibodies are useful for applications which include repeated administration to subjects. Chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, can be made using standard recombinant DNA techniques. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art, for example using methods described in Robinson et al International Application No. PCT/US86/02269; Akira, et al European Patent Application 184,187; Taniguchi, M., European Patent Application 171,496; Morrison et al European Patent Application 173,494; Neuberger et al PCT International Publication No. WO 86/01533; Cabilly et al U.S. Pat. No. 4,816,567; Cabilly et al European Patent Application 125,023; Better et al., Science 240: 1041-1043 (1988); Liu et al., Proc. Natl. Acad. Sci. USA 84: 3439-3443 (1987); Liu et al., J. Immunol. 139: 3521-3526 (1987); Sun et al., Proc. Natl. Acad. Sci. USA 84: 214-218 (1987); Nishimura et al., Canc. Res. 47: 999-1005 (1987); Wood et al., Nature 314: 446-449 (1985); and Shaw et al., J. Natl. Cancer Inst. 80: 1553-1559 (1988); Morrison, S. L., Science 229: 1202-1207 (1985); Oi et al., BioTechniques 4: 214 (1986); Winter U.S. Pat. No. 5,225,539; Jones et al., Nature 321: 552-525 (1986); Verhoeyan et al., Science 239: 1534; and Beidler et al., J. Immunol. 141: 4053-4060 (1988).


Completely human antibodies are particularly desirable for therapeutic treatment of human patients. Such antibodies can be produced using transgenic mice that are incapable of expressing endogenous immunoglobulin heavy and light chains genes, but which can express human heavy and light chain genes. See, for example, Lonberg and Huszar, Int. Rev. Immunol. 13: 65-93 (1995); and U.S. Pat. Nos. 5,625,126; 5,633,425; 5,569,825; 5,661,016; and 5,545,806. In addition, companies such as Abgenix, Inc. (Fremont, Calif.) and Medarex, Inc. (Princeton, N.J.), can be engaged to provide human antibodies directed against a selected antigen using technology similar to that described above. Completely human antibodies that recognize a selected epitope also can be generated using a technique referred to as “guided selection.” In this approach a selected non-human monoclonal antibody (e.g., a murine antibody) is used to guide the selection of a completely human antibody recognizing the same epitope. This technology is described for example by Jespers et al., Bio/Technology 12: 899-903 (1994).


An anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibody can be a single chain antibody. A single chain antibody (scFV) can be engineered (see, e.g., Colcher et al., Ann. NY Acad. Sci. 880: 263-80 (1999); and Reiter, Clin. Cancer Res. 2: 245-52 (1996)). Single chain antibodies can be dimerized or multimerized to generate multivalent antibodies having specificities for different epitopes of the same target NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide.


Antibodies also may be selected or modified so that they exhibit reduced or no ability to bind an Fc receptor. For example, an antibody may be an isotype or subtype, fragment or other mutant, which does not support binding to an Fc receptor (e.g., it has a mutagenized or deleted Fc receptor binding region).


Also, an antibody (or fragment thereof) may be conjugated to a therapeutic moiety such as a cytotoxin, a therapeutic agent or a radioactive metal ion. A cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Examples include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thiotepa chlorambucil, melphalan, carmustine (BCNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine and vinblastine).


Antibody conjugates can be used for modifying a given biological response. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a polypeptide such as tumor necrosis factor, gamma-interferon, alpha-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator; or, biological response modifiers such as, for example, lymphokines, interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors. Also, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Pat. No. 4,676,980, for example.


An anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibody (e.g., monoclonal antibody) can be used to isolate NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides by standard techniques, such as affinity chromatography or immunoprecipitation. Moreover, an anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibody can be used to detect a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression of the polypeptide. Anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibodies can be used diagnostically to monitor polypeptide levels in tissue as part of a clinical testing procedure, e.g., to determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance (i.e., antibody labeling). Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, 3-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 125I, 131I, 35S or 3H. Also, an anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibody can be utilized as a test molecule for determining whether it can treat melanoma, and as a therapeutic for administration to a subject for treating melanoma.


An antibody can be made by immunizing with a purified NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antigen, or a fragment thereof, e.g., a fragment described herein, a membrane associated antigen, tissues, e.g., crude tissue preparations, whole cells, preferably living cells, lysed cells, or cell fractions.


Included herein are antibodies which bind only a native NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, only denatured or otherwise non-native NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, or which bind both, as well as those having linear or conformational epitopes. Conformational epitopes sometimes can be identified by selecting antibodies that bind to native but not denatured NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Also featured are antibodies that specifically bind to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK PCLO or REPS2 protein or polypeptide variant associated with melanoma. For example, the antibody sometimes specifically binds to an epitope that comprises a serine at amino acid position 656 in a NID2 protein, polypeptide or peptide.


Screening Assays


Featured herein are methods for identifying a candidate therapeutic for treating melanoma and detecting occurrence of melanoma. The methods comprise contacting a test molecule with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, substantially identical nucleic acid, polypeptide, substantially identical polypeptide, or fragment of the foregoing in a system. The nucleic acid often is a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleotide sequence represented by SEQ ID NO: 1-17, respectively; sometimes is a nucleotide sequence substantially identical to the nucleotide sequence of SEQ ID NO: 1-17 or sometimes a fragment thereof; and the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or fragment thereof is a polypeptide encoded by any of these nucleic acids. The method also comprises determining the presence or absence of an interaction between the test molecule and the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or polypeptide, where the presence of an interaction between the test molecule and the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or polypeptide identifies the test molecule as a candidate melanoma therapeutic.


As used herein, the term “test molecule” and “candidate therapeutic” refers to modulators of regulation of transcription and translation of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids and modulations of expression and activity of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides. The term “modulator” as used herein refers to a molecule which agonizes or antagonizes NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 DNA replication and/or DNA processing (e.g., methylation), NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 RNA transcription and/or RNA processing (e.g., removal of intronic sequences and/or translocation from the nucleus), NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide production (e.g., translation of the polypeptide from mRNA, and/or post-translational modification such as glycosylation, phosphorylation, and proteolysis of pro-polypeptides), and/or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 function (e.g., conformational changes, binding of nucleotides or nucleotide analogs, interaction with binding partners, effect on phosphorylation, and/or effect on occurrence of melanoma). Test molecules and candidate therapeutics include, but are not limited to, compounds, RNAi or siRNA molecules, antisense nucleic acids, ribozymes, NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides or fragments thereof, and immunotherapeutics (e.g., antibodies and HLA-presented polypeptide fragments).


As used herein, the term “system” refers to a cell free in vitro environment and a cell-based environment such as a collection of cells, a tissue, an organ, or an organism. A system is “contacted” with a test molecule in a variety of manners, including adding molecules in solution and allowing them to interact with one another by diffusion, cell injection, and any administration routes in an animal. As used herein, the term “interaction” refers to an effect of a test molecule on a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, polypeptide, or variant thereof (collectively referred to as a “NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule”), where the effect is sometimes binding between the test molecule and the nucleic acid or polypeptide, and is often an observable change in cells, tissue, or organism.


There are many standard methods for detecting the presence or absence of interaction between a test molecule and a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or polypeptide. For example, titrametric, acidimetric, radiometric, NMR, monolayer, polarographic, spectrophotometric, fluorescent, and ESR assays probative of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 function may be utilized.


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity and/or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 interactions can be detected and quantified using assays known in the art and described in Examples hereafter.


An interaction can be determined by labeling the test molecule and/or the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule, where the label is covalently or non-covalently attached to the test molecule or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule. The label is sometimes a radioactive molecule such as 125I, 131I, 35S or 3H, which can be detected by direct counting of radioemission or by scintillation counting. Also, enzymatic labels such as horseradish peroxidase, alkaline phosphatase, or luciferase may be utilized where the enzymatic label can be detected by determining conversion of an appropriate substrate to product. Also, presence or absence of an interaction can be determined without labeling. For example, a microphysiometer (e.g., Cytosensor) is an analytical instrument that measures the rate at which a cell acidifies its environment using a light-addressable potentiometric sensor (LAPS). Changes in this acidification rate can be used as an indication of an interaction between a test molecule and NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 (McConnell, H. M. et al., Science 257: 1906-1912 (1992)).


In cell-based systems, cells typically include a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or polypeptide or variants thereof and are often of mammalian origin, although the cell can be of any origin. Whole cells, cell homogenates, and cell fractions (e.g., cell membrane fractions) can be subjected to analysis. Where interactions between a test molecule with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or variant thereof are monitored, soluble and/or membrane bound forms of the polypeptide or variant may be utilized. Where membrane-bound forms of the polypeptide are used, it may be desirable to utilize a solubilizing agent. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton® X-100, Triton® X-114, Thesit®, Isotridecypoly(ethylene glycol ether)n, 3-[(3-cholamidopropyl)dimethylamminio]-1-propane sulfonate (CHAPS), 3-[(3cholamidopropyl)dimethylamminio]-2-hydroxy-1-propane sulfonate (CHAPSO), or N-dodecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate.


An interaction between two molecules can also be detected by monitoring fluorescence energy transfer (FET) (see, for example, Lakowicz et al., U.S. Pat. No. 5,631,169; Stavrianopoulos et al. U.S. Pat. No. 4,868,103). A fluorophore label on a first, “donor” molecule is selected such that its emitted fluorescent energy will be absorbed by a fluorescent label on a second, “acceptor” molecule, which in turn is able to fluoresce due to the absorbed energy. Alternately, the “donor” polypeptide molecule may simply utilize the natural fluorescent energy of tryptophan residues. Labels are chosen that emit different wavelengths of light, such that the “acceptor” molecule label may be differentiated from that of the “donor”. Since the efficiency of energy transfer between the labels is related to the distance separating the molecules, the spatial relationship between the molecules can be assessed. In a situation in which binding occurs between the molecules, the fluorescent emission of the “acceptor” molecule label in the assay should be maximal. An FET binding event can be conveniently measured through standard fluorometric detection means well known in the art (e.g., using a fluorimeter).


In another embodiment, determining the presence or absence of an interaction between a test molecule and a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule can be effected by using real-time Biomolecular Interaction Analysis (BIA) (see, e.g., Sjolander & Urbaniczk, Anal. Chem. 63: 2338-2345 (1991) and Szabo et al., Curr. Opin. Struct. Biol. 5: 699-705 (1995)). “Surface plasmon resonance” or “BIA” detects biospecific interactions in real time, without labeling any of the interactants (e.g., BIAcore). Changes in the mass at the binding surface (indicative of a binding event) result in alterations of the refractive index of light near the surface (the optical phenomenon of surface plasmon resonance (SPR)), resulting in a detectable signal which can be used as an indication of real-time reactions between biological molecules.


In another embodiment, the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule or test molecules are anchored to a solid phase. The NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule/test molecule complexes anchored to the solid phase can be detected at the end of the reaction. The target NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule is often anchored to a solid surface, and the test molecule, which is not anchored, can be labeled, either directly or indirectly, with detectable labels discussed herein.


It may be desirable to immobilize a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule, an anti-NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibody, or test molecules to facilitate separation of complexed from uncomplexed forms of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules and test molecules, as well as to accommodate automation of the assay. Binding of a test molecule to a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtitre plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion polypeptide can be provided which adds a domain that allows a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule to be bound to a matrix. For example, glutathione-5-transferase/NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 fusion polypeptides or glutathione-5-transferase/target fusion polypeptides can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtitre plates, which are then combined with the test compound or the test compound and either the non-adsorbed target polypeptide or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, and the mixture incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads or microtitre plate wells are washed to remove any unbound components, the matrix immobilized in the case of beads, complex determined either directly or indirectly, for example, as described above. Alternatively, the complexes can be dissociated from the matrix, and the level of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 binding or activity determined using standard techniques.


Other techniques for immobilizing a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule on matrices include using biotin and streptavidin. For example, biotinylated NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or target molecules can be prepared from biotin-NHS(N-hydroxy-succinimide) using techniques known in the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical).


In order to conduct the assay, the non-immobilized component is added to the coated surface containing the anchored component. After the reaction is complete, unreacted components are removed (e.g., by washing) under conditions such that any complexes formed will remain immobilized on the solid surface. The detection of complexes anchored on the solid surface can be accomplished in a number of ways. Where the previously non-immobilized component is pre-labeled, the detection of label immobilized on the surface indicates that complexes were formed. Where the previously non-immobilized component is not pre-labeled, an indirect label can be used to detect complexes anchored on the surface; e.g., using a labeled antibody specific for the immobilized component (the antibody, in turn, can be directly labeled or indirectly labeled with, e.g., a labeled anti-Ig antibody).


In one embodiment, this assay is performed utilizing antibodies reactive with NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or test molecules but which do not interfere with binding of the NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide to its test molecule. Such antibodies can be derivatized to the wells of the plate, and unbound target or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or target molecule, as well as enzyme-linked assays which rely on detecting an enzymatic activity associated with the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or test molecule.


Alternatively, cell free assays can be conducted in a liquid phase. In such an assay, the reaction products are separated from unreacted components, by any of a number of standard techniques, including but not limited to: differential centrifugation (see, for example, Rivas, G., and Minton, A. P., Trends Biochem Sci August; 18(8): 284-7 (1993)); chromatography (gel filtration chromatography, ion-exchange chromatography); electrophoresis (see, e.g., Ausubel et al., eds. Current Protocols in Molecular Biology, J Wiley: New York (1999)); and immunoprecipitation (see, for example, Ausubel, F. et al., eds. Current Protocols in Molecular Biology, J. Wiley: New York (1999)). Such resins and chromatographic techniques are known to one skilled in the art (see, e.g., Heegaard, J. Mol. Recognit. Winter; 11(1-6): 141-8 (1998); Hage & Tweed, J. Chromatogr. B Biomed. Sci. Appl. October 10; 699 (1-2): 499-525 (1997)). Further, fluorescence energy transfer may also be conveniently utilized, as described herein, to detect binding without further purification of the complex from solution.


In another embodiment, modulators of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression are identified. For example, a cell or cell free mixture is contacted with a candidate compound and the expression of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide evaluated relative to the level of expression of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide in the absence of the candidate compound. When expression of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide is greater in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide expression. Alternatively, when expression of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide is less (statistically significantly less) in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide expression. The level of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide expression can be determined by methods described herein for detecting NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 mRNA or polypeptide.


In an embodiment, binding partners that interact with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule are detected. The NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules can interact with one or more cellular or extracellular macromolecules, such as polypeptides, in vivo, and these molecules that interact with NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules are referred to herein as “binding partners.” Molecules that disrupt such interactions can be useful in regulating the activity of the target gene product. Such molecules can include, but are not limited to molecules such as antibodies, peptides, and small molecules (e.g., siRNA). The preferred target genes/products for use in this embodiment are the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 genes herein identified. In an alternative embodiment, provided are methods for determining the ability of the test compound to modulate the activity of a NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide through modulation of the activity of a downstream effector of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 target molecule. For example, the activity of the effector molecule on an appropriate target can be determined, or the binding of the effector to an appropriate target can be determined, as previously described.


To identify compounds that interfere with the interaction between the target gene product and its cellular or extracellular binding partner(s), e.g., a substrate, a reaction mixture containing the target gene product and the binding partner is prepared, under conditions and for a time sufficient, to allow the two products to form complex. In order to test an inhibitory agent, the reaction mixture is provided in the presence and absence of the test compound. The test compound can be initially included in the reaction mixture, or can be added at a time subsequent to the addition of the target gene and its cellular or extracellular binding partner. Control reaction mixtures are incubated without the test compound or with a placebo. The formation of any complexes between the target gene product and the cellular or extracellular binding partner is then detected. The formation of a complex in the control reaction, but not in the reaction mixture containing the test compound, indicates that the compound interferes with the interaction of the target gene product and the interactive binding partner. Additionally, complex formation within reaction mixtures containing the test compound and normal target gene product can also be compared to complex formation within reaction mixtures containing the test compound and mutant target gene product. This comparison can be important in those cases wherein it is desirable to identify compounds that disrupt interactions of mutant but not normal target gene products.


These assays can be conducted in a heterogeneous or homogeneous format. Heterogeneous assays involve anchoring either the target gene product or the binding partner onto a solid phase, and detecting complexes anchored on the solid phase at the end of the reaction. In homogeneous assays, the entire reaction is carried out in a liquid phase. In either approach, the order of addition of reactants can be varied to obtain different information about the compounds being tested. For example, test compounds that interfere with the interaction between the target gene products and the binding partners, e.g., by competition, can be identified by conducting the reaction in the presence of the test substance. Alternatively, test compounds that disrupt preformed complexes, e.g., compounds with higher binding constants that displace one of the components from the complex, can be tested by adding the test compound to the reaction mixture after complexes have been formed. The various formats are briefly described below.


In a heterogeneous assay system, either the target gene product or the interactive cellular or extracellular binding partner, is anchored onto a solid surface (e.g., a microtitre plate), while the non-anchored species is labeled, either directly or indirectly. The anchored species can be immobilized by non-covalent or covalent attachments. Alternatively, an immobilized antibody specific for the species to be anchored can be used to anchor the species to the solid surface.


In order to conduct the assay, the partner of the immobilized species is exposed to the coated surface with or without the test compound. After the reaction is complete, unreacted components are removed (e.g., by washing) and any complexes formed will remain immobilized on the solid surface. Where the non-immobilized species is pre-labeled, the detection of label immobilized on the surface indicates that complexes were formed. Where the non-immobilized species is not pre-labeled, an indirect label can be used to detect complexes anchored on the surface; e.g., using a labeled antibody specific for the initially non-immobilized species (the antibody, in turn, can be directly labeled or indirectly labeled with, e.g., a labeled anti-Ig antibody). Depending upon the order of addition of reaction components, test compounds that inhibit complex formation or that disrupt preformed complexes can be detected.


Alternatively, the reaction can be conducted in a liquid phase in the presence or absence of the test compound, the reaction products separated from unreacted components, and complexes detected; e.g., using an immobilized antibody specific for one of the binding components to anchor any complexes formed in solution, and a labeled antibody specific for the other partner to detect anchored complexes. Again, depending upon the order of addition of reactants to the liquid phase, test compounds that inhibit complex or that disrupt preformed complexes can be identified.


In an alternate embodiment, a homogeneous assay can be used. For example, a preformed complex of the target gene product and the interactive cellular or extracellular binding partner product is prepared in that either the target gene products or their binding partners are labeled, but the signal generated by the label is quenched due to complex formation (see, e.g., U.S. Pat. No. 4,109,496 that utilizes this approach for immunoassays). The addition of a test substance that competes with and displaces one of the species from the preformed complex will result in the generation of a signal above background. In this way, test substances that disrupt target gene product-binding partner interaction can be identified.


Also, binding partners of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules can be identified in a two-hybrid assay or three-hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et al., Cell 72: 223-232 (1993); Madura et al., J. Biol. Chem. 268: 12046-12054 (1993); Bartel et al., Biotechniques 14: 920-924 (1993); Iwabuchi et al., Oncogene 8: 1693-1696 (1993); and Brent WO94/10300), to identify other polypeptides, which bind to or interact with NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 (“NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2-binding polypeptides” or “NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2-bp”) and are involved in NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity. Such NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2-bps can be activators or inhibitors of signals by the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 targets as, for example, downstream elements of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2-mediated signaling pathway.


A two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene that codes for a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4). In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified polypeptide (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor. (Alternatively the: NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide can be the fused to the activator domain.) If the “bait” and the “prey” polypeptides are able to interact, in vivo, forming a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., LacZ) which is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene which encodes the polypeptide which interacts with the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide.


Candidate therapeutics for treating melanoma are identified from a group of test molecules that interact with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid or polypeptide. Test molecules often are ranked according to the degree with which they interact or modulate (e.g., agonize or antagonize) DNA replication and/or processing, RNA transcription and/or processing, polypeptide production and/or processing, and/or function of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules, for example, and then top ranking modulators are selected. Also, pharmacogenomic information described herein can determine the rank of a modulator. Candidate therapeutics typically are formulated for administration to a subject.


Therapeutic Treatments


Formulations or pharmaceutical compositions typically include in combination with a pharmaceutically acceptable carrier a compound, an antisense nucleic acid, a ribozyme, an antibody, a binding partner that interacts with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid, or a fragment thereof. The formulated molecule may be one that is identified by a screening method described above. Also, formulations may comprise a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or fragment thereof and a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable carrier” includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Supplementary active compounds can also be incorporated into the compositions.


A pharmaceutical composition is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.


Oral compositions generally include an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.


Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.


Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation often utilized are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.


For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.


Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams (e.g., sunscreen) as generally known in the art. Molecules can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.


In one embodiment, active molecules are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. Materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.


It is advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.


Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50. Molecules which exhibit high therapeutic indices often are utilized. While molecules that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.


The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such molecules lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any molecules used in the method, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.


As defined herein, a therapeutically effective amount of protein or polypeptide (i.e., an effective dosage) ranges from about 0.001 to 30 mg/kg body weight, sometimes about 0.01 to 25 mg/kg body weight, often about 0.1 to 20 mg/kg body weight, and more often about 1 to 10 mg/kg, 2 to 9 mg/kg, 3 to 8 mg/kg, 4 to 7 mg/kg, or 5 to 6 mg/kg body weight. The protein or polypeptide can be administered one time per week for between about 1 to 10 weeks, sometimes between 2 to 8 weeks, often between about 3 to 7 weeks, and more often for about 4, 5, or 6 weeks. The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of a protein, polypeptide, or antibody can include a single treatment or, preferably, can include a series of treatments.


With regard to polypeptide formulations, featured herein is a method for treating melanoma in a subject, which comprises contacting one or more cells in the subject with a first NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide, where genomic DNA in the subject comprises a second NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid having one or more polymorphic variations associated with melanoma. The first NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide comprises fewer polymorphic variations associated with melanoma than the second NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. The first and second NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides are encoded by a nucleic acid which comprises a nucleotide sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1-17; a nucleotide sequence which encodes a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 18-27; and a nucleotide sequence which encodes a polypeptide that is 90% or more identical to an amino acid sequence set forth in SEQ ID NO: 18-27. The second NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide also may be encoded by a fragment of the foregoing nucleic acids comprising the one or more polymorphic variations. The subject is often a human.


For antibodies, a dosage of 0.1 mg/kg of body weight (generally 10 mg/kg to 20 mg/kg) is often utilized. If the antibody is to act in the brain, a dosage of 50 mg/kg to 100 mg/kg is often appropriate. Generally, partially human antibodies and fully human antibodies have a longer half-life within the human body than other antibodies. Accordingly, lower dosages and less frequent administration is often possible. Modifications such as lipidation can be used to stabilize antibodies and to enhance uptake and tissue penetration (e.g., into the brain). A method for lipidation of antibodies is described by Cruikshank et al., J. Acquired Immune Deficiency Syndromes and Human Retrovirology 14: 193 (1997).


Antibody conjugates can be used for modifying a given biological response, the drug moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a polypeptide such as tumor necrosis factor, .alpha.-interferon, .beta.-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator; or, biological response modifiers such as, for example, lymphokines, interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors. Alternatively, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Pat. No. 4,676,980.


For compounds, exemplary doses include milligram or microgram amounts of the compound per kilogram of subject or sample weight, for example, about 1 microgram per kilogram to about 500 milligrams per kilogram, about 100 micrograms per kilogram to about 5 milligrams per kilogram, or about 1 microgram per kilogram to about 50 micrograms per kilogram. It is understood that appropriate doses of a small molecule depend upon the potency of the small molecule with respect to the expression or activity to be modulated. When one or more of these small molecules is to be administered to an animal (e.g., a human) in order to modulate expression or activity of a polypeptide or nucleic acid, a physician, veterinarian, or researcher may, for example, prescribe a relatively low dose at first, subsequently increasing the dose until an appropriate response is obtained. In addition, it is understood that the specific dose level for any particular animal subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, gender, and diet of the subject, the time of administration, the route of administration, the rate of excretion, any drug combination, and the degree of expression or activity to be modulated.


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid molecules can be inserted into vectors and used in gene therapy methods for treating melanoma. Featured herein is a method for treating melanoma in a subject, which comprises contacting one or more cells in the subject with a first NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid. Genomic DNA in the subject comprises a second NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid comprising one or more polymorphic variations associated with melanoma, and the first NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid comprises fewer polymorphic variations associated with melanoma. The first and second NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids typically comprise a nucleotide sequence selected from the group consisting of the nucleotide sequence of SEQ ID NO: 1-17; a nucleotide sequence which encodes a polypeptide consisting of an amino acid sequence set forth in SEQ ID NO: 18-27; and a nucleotide sequence which encodes a polypeptide that is 90% or more identical to an amino acid sequence set forth in SEQ ID NO: 18-27. The second NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid may also be a fragment of the foregoing comprising one or more polymorphic variations. The subject is often a human.


Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see U.S. Pat. No. 5,328,470) or by stereotactic injection (see e.g., Chen et al., (1994) Proc. Natl. Acad. Sci. USA 91: 3054-3057). Pharmaceutical preparations of gene therapy vectors can include a gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells (e.g., retroviral vectors) the pharmaceutical preparation can include one or more cells which produce the gene delivery system. Examples of gene delivery vectors are described herein.


Pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.


Pharmaceutical compositions of active ingredients can be administered by any of the paths described herein for therapeutic and prophylactic methods for treating melanoma. With regard to both prophylactic and therapeutic methods of treatment, such treatments may be specifically tailored or modified, based on knowledge obtained from pharmacogenomic analyses described herein. As used herein, the term “treatment” is defined as the application or administration of a therapeutic agent to a patient, or application or administration of a therapeutic agent to an isolated tissue or cell line from a patient, who has a disease, a symptom of disease or a predisposition toward a disease, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease, the symptoms of disease or the predisposition toward disease. A therapeutic agent includes, but is not limited to, small molecules, peptides, antibodies, ribozymes and antisense oligonucleotides.


Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 aberrance, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending on the type of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 aberrance, for example, a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecule, NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 agonist, or NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein.


As discussed, successful treatment of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 disorders can be brought about by tech niques that serve to inhibit the expression or activity of target gene products. For example, compounds (e.g., an agent identified using an assays described above) that exhibit negative modulatory activity can be used to prevent and/or treat melanoma. Such molecules can include, but are not limited to peptides, phosphopeptides, small organic or inorganic molecules, or antibodies (including, for example, polyclonal, monoclonal, humanized, anti-idiotypic, chimeric or single chain antibodies, and FAb, F(ab′)2 and FAb expression library fragments, scFV molecules, and epitope-binding fragments thereof).


Further, antisense and ribozyme molecules that inhibit expression of the target gene can also be used to reduce the level of target gene expression, thus effectively reducing the level of target gene activity. Still further, triple helix molecules can be utilized in reducing the level of target gene activity. Antisense, ribozyme and triple helix molecules are discussed above.


It is possible that the use of antisense, ribozyme, and/or triple helix molecules to reduce or inhibit mutant gene expression can also reduce or inhibit the transcription (triple helix) and/or translation (antisense, ribozyme) of mRNA produced by normal target gene alleles, such that the concentration of normal target gene product present can be lower than is necessary for a normal phenotype. In such cases, nucleic acid molecules that encode and express target gene polypeptides exhibiting normal target gene activity can be introduced into cells via gene therapy method. Alternatively, in instances in that the target gene encodes an extracellular polypeptide, it can be preferable to co-administer normal target gene polypeptide into the cell or tissue in order to maintain the requisite level of cellular or tissue target gene activity.


Another method by which nucleic acid molecules may be utilized in treating or preventing a disease characterized by NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression is through the use of aptamer molecules specific for NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Aptamers are nucleic acid molecules having a tertiary structure which permits them to specifically bind to polypeptide ligands (see, e.g., Osborne, et al., Curr. Opin. Chem. Biol. 1 (1): 5-9 (1997); and Patel, D. J., Curr. Opin. Chem. Biol. June; 1 (1): 3246 (1997)). Since nucleic acid molecules may in many cases be more conveniently introduced into target cells than therapeutic polypeptide molecules may be, aptamers offer a method by which NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide activity may be specifically decreased without the introduction of drugs or other molecules which may have pluripotent effects.


Antibodies can be generated that are both specific for target gene product and that reduce target gene product activity. Such antibodies may, therefore, by administered in instances whereby negative modulatory techniques are appropriate for the treatment of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 disorders. For a description of antibodies, see the Antibody section above.


In circumstances where injection of an animal or a human subject with a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or epitope for stimulating antibody production is harmful to the subject, it is possible to generate an immune response against NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 through the use of anti-idiotypic antibodies (see, for example, Herlyn, D., Ann. Med.; 31(1): 66-78 (1999); and Bhattacharya-Chatterjee & Foon, Cancer Treat. Res.; 94: 51-68 (1998)). If an anti-idiotypic antibody is introduced into a mammal or human subject, it should stimulate the production of anti-anti-idiotypic antibodies, which should be specific to the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide. Vaccines directed to a disease characterized by NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression may also be generated in this fashion.


In instances where the target antigen is intracellular and whole antibodies are used, internalizing antibodies often are utilized. Lipofectin or liposomes can be used to deliver the antibody or a fragment of the Fab region that binds to the target antigen into cells. Where fragments of the antibody are used, the smallest inhibitory fragment that binds to the target antigen often are utilized. For example, peptides having an amino acid sequence corresponding to the Fv region of the antibody can be used. Alternatively, single chain neutralizing antibodies that bind to intracellular target antigens can also be administered. Such single chain antibodies can be administered, for example, by expressing nucleotide sequences encoding single-chain antibodies within the target cell population (see e.g., Marasco et al., Proc. Natl. Acad. Sci. USA 90: 7889-7893 (1993)).


NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules and compounds that inhibit target gene expression, synthesis and/or activity can be administered to a patient at therapeutically effective doses to prevent, treat or ameliorate NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 disorders. A therapeutically effective dose refers to that amount of the compound sufficient to result in amelioration of symptoms of the disorders.


Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50. Compounds that exhibit large therapeutic indices often are utilized. While compounds that exhibit toxic side effects can be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.


Data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography.


Another example of effective dose determination for an individual is the ability to directly assay levels of “free” and “bound” compound in the serum of the test subject. Such assays may utilize antibody mimics and/or “biosensors” that have been created through molecular imprinting techniques.


The compound which is able to modulate NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is used as a template, or “imprinting molecule”, to spatially organize polymerizable monomers prior to their polymerization with catalytic reagents. The subsequent removal of the imprinted molecule leaves a polymer matrix which contains a repeated “negative image” of the compound and is able to selectively rebind the molecule under biological assay conditions. A detailed review of this technique can be seen in Ansell et al., Current Opinion in Biotechnology 7: 89-94 (1996) and in Shea, Trends in Polymer Science 2: 166-173 (1994). Such “imprinted” affinity matrixes are amenable to ligand-binding assays, whereby the immobilized monoclonal antibody component is replaced by an appropriately imprinted matrix. An example of the use of such matrixes in this way can be seen in Vlatakis, et al., Nature 361: 645-647 (1993). Through the use of isotope-labeling, the “free” concentration of compound which modulates the expression or activity of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 can be readily monitored and used in calculations of IC50. Such “imprinted” affinity matrixes can also be designed to include fluorescent groups whose photon-emitting properties measurably change upon local and selective binding of target compound. These changes can be readily assayed in real time using appropriate fiberoptic devices, in turn allowing the dose in a test subject to be quickly optimized based on its individual IC50. A rudimentary example of such a “biosensor” is discussed in Kriz et al., Analytical Chemistry 67: 2142-2144 (1995).


Provided herein are methods of modulating NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression or activity for therapeutic purposes. Accordingly, in an exemplary embodiment, the modulatory method involves contacting a cell with NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 or an agent that modulates one or more activities of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide activity associated with the cell. An agent that modulates NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide activity can be an agent as described herein, such as a nucleic acid or a polypeptide, a naturally-occurring target molecule of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide (e.g., a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 substrate or receptor), a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibody, a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 agonist or antagonist, a peptidomimetic of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 agonist or antagonist, or other small molecule.


In one embodiment, the agent stimulates one or more NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activities. Examples of such stimulatory agents include active NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide and a nucleic acid molecule encoding NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2. In another embodiment, the agent inhibits one or more NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activities. Examples of such inhibitory agents include antisense NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acid molecules, anti-NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 antibodies, and NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 inhibitors. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, provided are methods of treating an individual afflicted with a disease or disorder characterized by aberrant or unwanted expression or activity of a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or nucleic acid molecule. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., upregulates or downregulates) NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression or activity. In another embodiment, the method involves administering a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptide or nucleic acid molecule as therapy to compensate for reduced, aberrant, or unwanted NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 expression or activity.


Stimulation of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is desirable in situations in which NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 is abnormally downregulated and/or in which increased NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is likely to have a beneficial effect. For example, stimulation of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is desirable in situations in which a NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 is downregulated and/or in which increased NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is likely to have a beneficial effect. Likewise, inhibition of NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is desirable in situations in which NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 is abnormally upregulated and/or in which decreased NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 activity is likely to have a beneficial effect.


The examples set forth below are intended to illustrate but not limit the invention.


EXAMPLES

In the following studies a group of subjects were selected according to specific parameters relating to melanoma. Nucleic acid samples obtained from individuals in the study group were subjected to genetic analysis, which identified associations between melanoma and certain polymorphic regions in the NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 genes. Methods are described for producing NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 polypeptides and polypeptide variants in vitro or in vivo. NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 nucleic acids or polypeptides and variants thereof are utilized for screening test molecules for those that interact with NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules. Test molecules identified as interactors with NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO or REPS2 molecules and variants are further screened in vivo to determine whether they treat melanoma.


Example 1
Samples and Pooling Strategies

Sample Selection


Blood samples were collected from individuals diagnosed with melanoma, which were referred to as case samples. Also, blood samples were collected from individuals not diagnosed with melanoma or a history of melanoma; these samples served as gender and age-matched controls. A database was created that listed all phenotypic trait information gathered from individuals for each case and control sample. Genomic DNA was extracted from each of the blood samples for genetic analyses.


DNA Extraction from Blood Samples


Six to ten milliliters of whole blood was transferred to a 50 ml tube containing 27 ml of red cell lysis solution (RCL). The tube was inverted until the contents were mixed. Each tube was incubated for 10 minutes at room temperature and inverted once during the incubation. The tubes were then centrifuged for 20 minutes at 3000×g and the supernatant was carefully poured off. 100-200 μl of residual liquid was left in the tube and was pipetted repeatedly to resuspend the pellet in the residual supernatant. White cell lysis solution (WCL) was added to the tube and pipetted repeatedly until completely mixed. While no incubation was normally required, the solution was incubated at 37° C. or room temperature if cell clumps were visible after mixing until the solution was homogeneous. Two ml of protein precipitation was added to the cell lysate. The mixtures were vortexed vigorously at high speed for 20 sec to mix the protein precipitation solution uniformly with the cell lysate, and then centrifuged for 10 minutes at 3000×g. The supernatant containing the DNA was then poured into a clean 15 ml tube, which contained 7 ml of 100% isopropanol. The samples were mixed by inverting the tubes gently until white threads of DNA were visible. Samples were centrifuged for 3 minutes at 2000×g and the DNA was visible as a small white pellet. The supernatant was decanted and 5 ml of 70% ethanol was added to each tube. Each tube was inverted several times to wash the DNA pellet, and then centrifuged for 1 minute at 2000×g. The ethanol was decanted and each tube was drained on clean absorbent paper. The DNA was dried in the tube by inversion for 10 minutes, and then 1000 μl of 1×TE was added. The size of each sample was estimated, and less TE buffer was added during the following DNA hydration step if the sample was smaller. The DNA was allowed to rehydrate overnight at room temperature, and DNA samples were stored at 2-8° C.


DNA was quantified by placing samples on a hematology mixer for at least 1 hour. DNA was serially diluted (typically 1:80, 1:160, 1:320, and 1:640 dilutions) so that it would be within the measurable range of standards. 125 μl of diluted DNA was transferred to a clear U-bottom microtitre plate, and 125 μl of 1×TE buffer was transferred into each well using a multichannel pipette. The DNA and 1×TE were mixed by repeated pipetting at least 15 times, and then the plates were sealed. 50 μl of diluted DNA was added to wells A5-H12 of a black flat bottom microtitre plate. Standards were inverted six times to mix them, and then 50 μl of 1×TE buffer was pipetted into well A1, 1000 ng/ml of standard was pipetted into well A2, 500 ng/ml of standard was pipetted into well A3, and 250 ng/ml of standard was pipetted into well A4. PicoGreen (Molecular Probes, Eugene, Oreg.) was thawed and freshly diluted 1:200 according to the number of plates that were being measured. PicoGreen was vortexed and then 501 μl was pipetted into all wells of the black plate with the diluted DNA. DNA and PicoGreen were mixed by pipetting repeatedly at least 10 times with the multichannel pipette. The plate was placed into a Fluoroskan Ascent Machine (microplate fluorometer produced by Labsystems) and the samples were allowed to incubate for 3 minutes before the machine was run using filter pairs 485 nm excitation and 538 nm emission wavelengths. Samples having measured DNA concentrations of greater than 450 ng/μl were re-measured for conformation. Samples having measured DNA concentrations of 20 ng/μl or less were re-measured for confirmation.


Pooling Strategies


Samples were placed into one of four groups, based on gender and disease status. The four groups were male case samples, male control samples, female case samples, and female control samples. A select set of samples from each group were utilized to generate pools, and one pool was created for each group. Each individual sample in a pool was represented by an equal amount of genomic DNA. For example, where 25 ng of genomic DNA was utilized in each PCR reaction and there were 200 individuals in each pool, each individual would provide 125 pg of genomic DNA. Inclusion or exclusion of samples for a pool was based upon the following criteria: the sample was derived from an individual characterized as Caucasian; the sample was derived from an individual of German paternal and maternal descent; the database included relevant phenotype information for the individual; case samples were derived from individuals diagnosed with melanoma; control samples were derived from individuals free of cancer; and sufficient genomic DNA was extracted from each blood sample for all allelotyping and genotyping reactions performed during the study. Phenotype information included sex of the individual, number of nevi (few, moderate, numerous), hair color (black, brown, blond, red), diagnosed with melanoma (tumor thickness, date of primary diagnosis, age of individual as of primary diagnosis, post-operative tumor classification, presence of nodes, occurrence of metastases, subtype, location), country or origin of mother and father, presence of certain conditions for each individual (coronary heart disease, cardiomyopathy, arteriosclerosis, abnormal blood clotting/thrombosis, emphysema, asthma, diabetes type 1, diabetes type 2, Alzheimer's disease, epilepsy, schizophrenia, manic depression/bipolar disorder, autoimmune disease, thyroid disorder, and hypertension), presence of cancer in the donor individual or blood relative (melanoma, basaliom/spinaliom/lentigo malignant/mycosis fungoides, breast cancer, colon cancer, rectum cancer, lung cancer, lung cancer, bronchus cancer, prostate cancer, stomach cancer, leukemia, lymphoma, or other cancer in donor, donor parent, donor aunt or uncle, donor offspring or donor grandparent. Samples that met these criteria were added to appropriate pools based on gender and disease status.


The selection process yielded the pools set forth in Table 1, which were used in the studies that follow:

TABLE 1MaleMaleFemaleFemalecontrolcasecontrolcasePool size (Number)217236233266Pool Criteriacontrolcasecontrolcase(ex: case/control)Mean Age (ex: years)48514749


Example 2
Association of Polymorphic Variants with Melanoma

A whole-genome screen was performed to identify particular SNPs associated with occurrence of melanoma. As described in Example 1, two sets of samples were utilized: female individuals having melanoma (female cases) and samples from female individuals not having melanoma or any history of melanoma (female controls), and male individuals having melanoma (male cases) and samples from male individuals not having melanoma or any history of melanoma (male controls). The initial screen of each pool was performed in an allelotyping study, in which certain samples in each group were pooled. By pooling DNA from each group, an allele frequency for each SNP in each group was calculated. These allele frequencies were then compared to one another. Particular SNPs were considered as being associated with melanoma when allele frequency differences calculated between case and control pools were statistically significant. SNP disease association results obtained from the allelotyping study were then validated by genotyping each associated SNP across all samples from each pool. The results of the genotyping were then analyzed, allele frequencies for each group were calculated from the individual genotyping results, and a p-value was calculated to determine whether the case and control groups had statistically significantly differences in allele frequencies for a particular SNP. When the genotyping results agreed with the original allelotyping results, the SNP disease association was considered validated at the genetic level.


SNP Panel Used for Genetic Analyses


A whole-genome SNP screen began with an initial screen of approximately 25,000 SNPs over each set of disease and control samples using a pooling approach. The pools studied in the screen are described in Example 1. The SNPs analyzed in this study were part of a set of 25,488 SNPs confirmed as being statistically polymorphic as each is characterized as having a minor allele frequency of greater than 10%. The SNPs in the set reside in genes or in close proximity to genes, and many reside in gene exons. Specifically, SNPs in the set are located in exons, introns, and within 5,000 base-pairs upstream of a transcription start site of a gene. In addition, SNPs were selected according to the following criteria: they are located in ESTs; they are located in Locuslink or Ensembl genes; and they are located in Genomatix promoter predictions. SNPs in the set were also selected on the basis of even spacing across the genome, as depicted in Table 2.

TABLE 2General StatisticsSpacing StatisticsTotal # of SNPs28,532Median34,424bp# of Exonic SNPs 8,497 (30%)Minimum*1,000bp# SNPs with26,625 (93%)Maximum*3,000,000bprefSNP IDGene Coverage23,874Mean122,412bpChromosomeAllStd354,bpCoverageDeviation
*Excludes outliers


Genotyping Results


The genetic studies summarized above and described in more detail below identified allelic variants associated with melanoma. The allelic variants identified from the SNP panel described in Table 2 are summarized below in Table 3.

TABLE 3MelanomaSNPChromo-ContigContigSequenceSequenceAllelicAssociatedReferencesomeIdentificationPositionIdentificationPositionVariabilityAllelers136045710NT_00886215592254NM_003873intronicT/CTrs75305014NT_02643732425413NM_007361intronicA/GArs72964717NT_01094019400937NM_006039intronicA/GGrs84041689506442NT_0105421320117NM_003674UTRC/Trs1044639782063609NT_0079337685796XM_168530UTRC/Ars2034453174082116NT_03297736275269NM_015978intronicA/Grs20344531NT_032977NM_003838intragenicrs1904528X12970302NT_01175712970302NM_004726UTRC/T


Table 3 includes information pertaining to the incident polymorphic variant associated with melanoma identified herein. Public information pertaining to the polymorphism and the genomic sequence that includes the polymorphism are indicated. The genomic sequence identified in Table 3 may be accessed at the http address www.ncbi.nih.gov/entrez/query.fcgi, for example, by using the publicly available SNP reference number (e.g., rs1360457). The chromosome position refers to the position of the SNP within NCBI's Genome Build 34 and dbSNP 119, which may be accessed at the following http address: www.ncbi.nlm.nih.gov/mapview/map_search.cgi?chr=hum_chr.inf&query=. The “Contig Position” provided in Table 3 corresponds to a nucleotide position set forth in the contig sequence, and designates the polymorphic site corresponding to the SNP reference number. The sequence containing the polymorphisms also may be referenced by the “Sequence Identification” set forth in Table 3. The “Sequence Identification” corresponds to cDNA sequence that encodes associated target polypeptides (e.g., NRP1 of the invention. The position of the SNP within the cDNA sequence is provided in the “Sequence Position” column of Table 3. In the case of rs2034453, genetic evidence suggests an association with a region on chromosome 1 band p31.1 that includes two genes: fucose-1-phosphate guanylyltransferase (FPGT) (NM003838) and cardiac ankyrin repeat kinase (CARK) (NM015978). Also, the allelic variation at the polymorphic site and the allelic variant identified as associated with melanoma is specified in Table 3. All nucleotide sequences referenced and accessed by the parameters set forth in Table 3 are incorporated herein by reference.


Assay for Verifying, Allelotyping, and Genotyping SNPs


A MassARRAY™ system (Sequenom, Inc.) was utilized to perform SNP genotyping in a high-throughput fashion. This genotyping platform was complemented by a homogeneous, single-tube assay method (hME™ or homogeneous MassEXTEND™ (Sequenom, Inc.)) in which two genotyping primers anneal to and amplify a genomic target surrounding a polymorphic site of interest. A third primer (the MassEXTEND™ primer), which is complementary to the amplified target up to but not including the polymorphism, was then enzymatically extended one or a few bases through the polymorphic site and then terminated.


For each polymorphism, SpectroDESIGNER™ software (Sequenom, Inc.) was used to generate a set of PCR primers and a MassEXTEND™ primer which where used to genotype the polymorphism. Other primer design software could be used or one of ordinary skill in the art could manually design primers based on his or her knowledge of the relevant factors and considerations in designing such primers. Table 4 shows PCR primers and Table 5 shows extension primers used for analyzing the polymorphism set forth in Table 3. The initial PCR amplification reaction was performed in a 5 μl total volume containing 1×PCR buffer with 1.5 mM MgCl2 (Qiagen), 200 μM each of dATP, dGTP, dCTP, dTTP (Gibco-BRL), 2.5 ng of genomic DNA, 0.1 units of HotStar DNA polymerase (Qiagen), and 200 nM each of forward and reverse PCR primers specific for the polymorphic region of interest.

TABLE 4PCR PrimersSNPReferenceForward PCR primerReverse PCR primerrs1360457TTGTCCCAACTGAGGCTTTGAACAGCCTTTCAGCTTTGGCrs753050ACGTTGGATGCCTCTGTTTCCAACTCAAGGACGTTGGATGTTCACAGAGGTTACTAAGGGrs729647AACCAACCCACTGGGTTGACTGATCTGGGAGAGTTTGGAGrs8404GCCTCCTGTTGGGTCCTCAAGGTATGGGGTGGGAGCrs1044639CGCAAACAAAAAGGACACACCTCCTTTGTTTCCACCATCCrs2034453TTGCTGGACAATAGAAAGACGTGACTGGAAACTGAGAATGrs1904528GAAGACTGAAAAAAATCCACGGCTATCTCTTTCACATTGCTC


Samples were incubated at 95° C. for 15 minutes, followed by 45 cycles of 95° C. for 20 seconds, 56° C. for 30 seconds, and 72° C. for 1 minute, finishing with a 3 minute final extension at 72° C. Following amplification, shrimp alkaline phosphatase (SAP) (0.3 units in a 2 μl volume) (Amersham Pharmacia) was added to each reaction (total reaction volume was 7 μl) to remove any residual dNTPs that were not consumed in the PCR step. Samples were incubated for 20 minutes at 37° C., followed by 5 minutes at 85° C. to denature the SAP.


Once the SAP reaction was complete, a primer extension reaction was initiated by adding a polymorphism-specific MassEXTEND™ primer cocktail to each sample. Each MassEXTEND™ cocktail included a specific combination of dideoxynucleotides (ddNTPs) and deoxynucleotides (dNTPs) used to distinguish polymorphic alleles from one another. Methods for verifying, allelotyping and genotyping SNPs are disclosed, for example, in U.S. Pat. No. 6,258,538, the content of which is hereby incorporated by reference. In Table 5, ddNTPs are shown and the fourth nucleotide not shown is the dNTP.

TABLE 5Extension PrimersSNPTerminationReferenceExtend ProbeMixrs1360457CAGCTTTGGCCAGGAGATGACTrs753050AGCACATAACACAGCATGGCACTrs729647GGTTGACGTCAACACAGGCACTrs8404CGAGACTACCAGGAGAGCCCACGrs1044639CATCCATCCAACCTGGCTCCGTrs2034453AAACTGAGAATGTTGATGGACAACTrs1904528TCACATTGCTCTGCACTTTTGACG


The MassEXTEND™ reaction was performed in a total volume of 9 μl, with the addition of 1× ThermoSequenase buffer, 0.576 units of ThermoSequenase (Amersham Pharmacia), 600 nM MassEXTEND™ primer, 2 mM of ddATP and/or ddCTP and/or ddGTP and/or ddTTP, and 2 mM of dATP or dCTP or dGTP or dTTP. The deoxy nucleotide (dNTP) used in the assay normally was complementary to the nucleotide at the polymorphic site in the amplicon. Samples were incubated at 94° C. for 2 minutes, followed by 55 cycles of 5 seconds at 94° C., 5 seconds at 52° C., and 5 seconds at 72° C.


Following incubation, samples were desalted by adding 16 μl of water (total reaction volume was 25 μl), 3 mg of SpectroCLEAN™ sample cleaning beads (Sequenom, Inc.) and allowed to incubate for 3 minutes with rotation. Samples were then robotically dispensed using a piezoelectric dispensing device (SpectroJET™ (Sequenom, Inc.)) onto either 96-spot or 384-spot silicon chips containing a matrix that crystallized each sample (SpectroCHIP™ (Sequenom, Inc.)). Subsequently, MALDI-TOF mass spectrometry (Biflex and Autoflex MALDI-TOF mass spectrometers (Bruker Daltonics) can be used) and SpectroTYPER RT™ software (Sequenom, Inc.) were used to analyze and interpret the SNP genotype for each sample.


Genetic Analysis


The minor allelic frequencies for the polymorphisms set forth in Table 3 were verified as being 10% or greater using the extension assay described above in a group of samples isolated from 92 individuals originating from the state of Utah in the United States, Venezuela and France (Coriell cell repositories).


Genotyping results for the allelic variant set forth in Table 3 are shown for female pools in Table 6 and for male pools in Table 8. In Table 6, “F case” and “F control” refer to female case and female control groups, and in Table 7, “M case” and “M control” refer to male case and male control groups.

TABLE 6Female Genotyping ResultsSNPp-OddsReferenceF caseF controlvalueRatiors1360457C = 0.869C = 0.9030.0890.71T = 0.131T = 0.097rs753050A = 0.270A = 0.2100.0210.71G = 0.730G = 0.790rs729647A = 0.578A = 0.6200.1871.19G = 0.422G = 0.380rs8404C = 0.144C = 0.0940.0160.62T = 0.856T = 0.906rs1044639A = 0.614A = 0.5280.0061.42C = 0.386C = 0.472rs2034453G = 0.854G = 0.9010.0230.64A = 0.146A = 0.099rs1904528T = 0.771T = 0.8380.0080.65C = 0.229C = 0.162









TABLE 7










Male Genotyping Results













SNP


p-
Odds



Reference
M case
M control
value
Ratio







rs1360457
C = 0.876
C = 0.917
0.042
0.66




T = 0.124
T = 0.083



rs753050
A = 0.270
A = 0.190
0.009
0.66




G = 0.730
G = 0.810



rs729647
A = 0.543
A = 0.640
0.004
1.48




G = 0.457
G = 0.360



rs8404
C = 0.136
C = 0.098
0.071
0.69




T = 0.864
T = 0.902



rs1044639
A = 0.611
A = 0.551
0.072
1.28




C = 0.389
C = 0.449



rs2034453
G = 0.896
G = 0.890
0.767
1.07




A = 0.104
A = 0.110



rs1904528
T = 0.790
T = 0.822
0.396
0.82




C = 0.210
C = 0.178










The single marker alleles set forth in Table 3 were considered validated, since the genotyping data for the females, males or both pools were significantly associated with melanoma, and because the genotyping results agreed with the original allelotyping results. Particularly significant associations with melanoma are indicated by a calculated p-value of less than 0.05 for genotype results, which are set forth in bold text.


The odds ratios were calculated for the alleles at each SNP and the results are shown in Tables 6 and 7. An odds ratio is an unbiased estimate of relative risk which can be obtained from most case-control studies. Relative risk (RR) is an estimate of the likelihood of disease in the exposed group (susceptibility allele or genotype carriers) compared to the unexposed group (not carriers). It can be calculated by the following equation:

RR═IA/Ia

    • IA is the incidence of disease in the A carriers and Ia is the incidence of disease in the non-carriers.
    • RR>1 indicates the A allele increases disease susceptibility.
    • RR<1 indicates the a allele increases disease susceptibility.


      For example, RR=1.5 indicates that carriers of the A allele have 1.5 times the risk of disease than non-carriers, i.e., 50% more likely to get the disease.


Case-control studies do not allow the direct estimation of IA and Ia, therefore relative risk cannot be directly estimated. However, the odds ratio (OR) can be calculated using the following equation:

OR=(nDAnda)/(ndAnDa)=pDA(1−pdA)/pdA(1−pDA), or
OR=((case f)/(1−case f))/((control f)/(1−control f)), where f=susceptibility allele frequency.


An odds ratio can be interpreted in the same way a relative risk is interpreted and can be directly estimated using the data from case-control studies, i.e., case and control allele frequencies. The higher the odds ratio value, the larger the effect that particular allele has on the development of melanoma, thus possessing that particular allele translates to having a higher risk of developing melanoma.


NID2 Non-Synonomous SNP


In addition, a non-synonomous, coding SNP (rs3742536) located at amino acid position 656 (P656S) of NID2 was genotyped and found to be significantly associated with the occurrence of melanoma. This SNP (rs3742536) was genotyped as described in Examples 1 and 2 herein using the following PCR primers, extend primer and termination mix: forward primer—ACGTTGGATGAGACCAACATTCAAGGCCAG; reverse primer—ACGTTGGATGAGTGGTACAGCTCCTTGTAG; extend primer—GGCTGTGAAATTTGCTG; and termination mix-ACT. The genotyping results for female (F) and male (M) cases and controls are shown in Table 8.

TABLE 8Genotpying ResultsChromosomeOddsRs numberPositionSexAllelesCase AFControl AFp-ValueRatiors374253650497467*MaleC/TT = 0.261T = 0.1920.01440.67C = 0.739C = 0.808FemaleT = 0.257T = 0.2050.05610.75C = 0.743C = 0.795
*based on NCBI's Build 34


Example 3

Samples and Pooling Strategies for the Replication Cohort


The single marker alleles set forth in Table 3 were genotyped again in a replication cohort to further validate their association with melanoma. Like the original study population described in Examples 1 and 2, the replication cohort consisted of individuals diagnosed with melanoma (cases) and individuals free of melanoma (controls). The case and control samples were selected and genotyped as described below.


Sample Selection


Blood samples were collected from individuals diagnosed with melanoma, which were referred to case samples. Also, blood samples were collected from individuals not diagnosed with me lanoma or a history of melanoma; these samples served as gender and age-matched controls.


DNA Extraction from Blood Samples


Blood samples for DNA preparation were taken in 5 EDTA tubes. If it was not possible to get a blood sample from a patient, a sample from the cheek mucosa was taken. Red blood cells were lysed to facilitate their separation from the white blood cells. The white cells were pelleted and lysed to release the DNA. Lysis was done in the presence of a DNA preservative using an anionic detergent to solubilize the cellular components. Contaminating RNA was removed by treatment with an RNA digesting enzyme. Cytoplasmic and nuclear proteins were removed by salt precipitation.


Genomic DNA was then isolated by precipitation with alcohol (2-propanol and then ethanol) and rehydrated in water. The DNA was transferred to 2-ml tubes and stored at 4° C. for short-term storage and at −70° C. for long-term storage.


Pooling Strategies


Samples were placed into one of four groups based on disease status. The four groups were female case samples, female control samples, male case samples, and male control samples. A select set of samples from each group were utilized to generate pools, and one pool was created for each group.


Replication samples were obtained from QIMR (Queensland Institute of Medical Research) through Nick Martin. All samples are of Australian descent. Sample sources were as follows: a.) Queensland Familial Melanoma Study-702 cutaneous malignant melanoma cases plus 46 unaffected relatives; and b.) Twin mole study-2367 controls, consisting of adolescent twins and siblings closest in age that had nevi counted and other skin and pigmentary phenotypes assessed. The subjects available for replication from Australia included 702 mostly unrelated melanoma cases from the Queensland Familial Melanoma study and 2367 controls from the Twin Mole study with subjects organized into pedigrees with up to eight unaffected individuals.


To facilitate a direct comparison with the discovery study design and subsequent meta analyses, we selected a subset of the Australian sample to produce a research data set of unrelated cases and controls. For cases, this was done by selecting the proband from all families within the familial melanoma study. For controls, this was accomplished by selecting the pedigree founders from the twin study, usually the father and mother of the collected twins. The resulting data set consisted of 376 female and 300 male cases, and 640 female and 515 male controls.


Example 4
Association of Polymorphic Variants with Melanoma in the Replication Cohort

The associated SNPs from the initial scan were re-validated by genotyping the associated SNP across the replication cohort described in Example 3. The results of the genotyping were then analyzed, allele frequencies for each group were calculated from the individual genotyping results, and a p-value was calculated to determine whether the case and control groups had statistically significantly differences in allele frequencies for a particular SNP. The replication genotyping results were considered significant with a calculated p-value of less than 0.05 for genotype results, which are set forth in bold text. See Tables 8 and 9 herein.


Assay for Verifying Allelotyping, and Genotyping SNPs


Genotyping of the replication cohort was performed using the same methods used for the original genotyping, as described herein. A MassARRAY™ system (Sequenom, Inc.) was utilized to perform SNP genotyping in a high-throughput fashion. This genotyping platform was complemented by a homogeneous, single-tube assay method (hME™ or homogeneous MassEXTEND™ (Sequenom, Inc.)) in which two genotyping primers anneal to and amplify a genomic target surrounding a polymorphic site of interest. A third primer (the MassEXTEND™ primer), which is complementary to the amplified target up to but not including the polymorphism, was then enzymatically extended one or a few bases through the polymorphic site and then terminated.


For each polymorphism, SpectroDESIGNER™ software (Sequenom, Inc.) was used to generate a set of PCR primers and a MassEXTEND™ primer which where used to genotype the polymorphism. Other primer design software could be used or one of ordinary skill in the art could manually design primers based on his or her knowledge of the relevant factors and considerations in designing such primers. Table 4 shows PCR primers and Table 5 shows extension probes used for analyzing (e.g., genotyping) polymorphisms. The initial PCR amplification reaction was performed in a 5 μl total volume containing 1×PCR buffer with 1.5 mM MgCl2 (Qiagen), 200 μM each of dATP, dGTP, dCTP, dTTP (Gibco-BRL), 2.5 ng of genomic DNA, 0.1 units of HotStar DNA polymerase (Qiagen), and 200 nM each of forward and reverse PCR primers specific for the polymorphic region of interest.


Samples were incubated at 95° C. for 15 minutes, followed by 45 cycles of 95° C. for 20 seconds, 56° C. for 30 seconds, and 72° C. for 1 minute, finishing with a 3 minute final extension at 72° C. Following amplification, shrimp alkaline phosphatase (SAP) (0.3 units in a 2 μl volume) (Amersham Pharmacia) was added to each reaction (total reaction volume was 7 μl) to remove any residual dNTPs that were not consumed in the PCR step. Samples were incubated for 20 minutes at 37° C., followed by 5 minutes at 85° C. to denature the SAP.


Once the SAP reaction was complete, a primer extension reaction was initiated by adding a polymorphism-specific MassEXTEND™ primer cocktail to each sample. Each MassEXTEND™ cocktail included a specific combination of dideoxynucleotides (ddNTPs) and deoxynucleotides (dNTPs) used to distinguish polymorphic alleles from one another. Methods for verifying, allelotyping and genotyping SNPs are disclosed, for example, in U.S. Pat. No. 6,258,538, the content of which is hereby incorporated by reference. In Table 5, ddNTPs are shown and the fourth nucleotide not shown is the dNTP.


The MassEXTEND™ reaction was performed in a total volume of 9 μl, with the addition of 1× ThermoSequenase buffer, 0.576 units of ThermoSequenase (Amersham Pharmacia), 600 nM MassEXTEND™ primer, 2 mM of ddATP and/or ddCTP and/or ddGTP and/or ddTTP, and 2 mM of dATP or dCTP or dGTP or dTTP. The deoxy nucleotide (dNTP) used in the assay normally was complementary to the nucleotide at the polymorphic site in the amplicon. Samples were incubated at 94° C. for 2 minutes, followed by 55 cycles of 5 seconds at 94° C., 5 seconds at 52° C., and 5 seconds at 72° C.


Following incubation, samples were desalted by adding 16 μl of water (total reaction volume was 25 μl), 3 mg of SpectroCLEAN™ sample cleaning beads (Sequenom, Inc.) and allowed to incubate for 3 minutes with rotation. Samples were then robotically dispensed using a piezoelectric dispensing device (SpectroJET™ (Sequenom, Inc.)) onto either 96-spot or 384-spot silicon chips containing a matrix that crystallized each sample (SpectroCHIP™ (Sequenom, Inc.)). Subsequently, MALDI-TOF mass spectrometry (Biflex and Autoflex MALDI-TOF mass spectrometers (Bruker Daltonics) can be used) and SpectroTYPER RT™ software (Sequenom, Inc.) were used to analyze and interpret the SNP genotype for each sample.


Genetic Analysis


The minor allelic frequencies for the polymorphisms set forth in Table 3 were verified as being 10% or greater using the extension assay described above in a group of samples isolated from 92 individuals originating from the state of Utah in the United States, Venezuela and France (Coriell cell repositories).


Replication genotyping results are shown for females and males in Table 9. P-values and odds ratios are provided in Table 10.

TABLE 9Female and Male Replication Genotyping ResultsAlleleAllelefrequency-femalesfrequency-malesdbSNPCon-Con-rs#GeneCasetrolDeltaCasetrolDeltars8404CDK100.870.91−0.0430.880.91−0.029rs1044639PCLO0.600.570.0310.610.560.042rs2034453FPGT/0.8587.00−0.0220.840.87−0.026TNNI3K(CARK)rs1904528REPS20.780.79−0.0050.770.82−0.051









TABLE 10










Female and Male Replication Genotyping Analysis











Replication F
Replication M
Summary














dbSNP

p-

p-

p-



rs#
Gene
value
OR
value
OR
value
OR





rs8404
CDK10
0.002
0.65
0.054
0.72
0.000004
0.67


rs1044639
PCLO
0.180
1.13
0.100
1.19
0.000290
1.23


rs2034453
FPGT/
0.170
0.83
0.150
0.81
0.014000
0.82



TNNI3K



(CARK)


rs1904528
REPS2
0.800
0.97
0.076
0.73
0.033000
0.80









Meta analyses, combining the results of the German discovery sample and the Australian replication sample, were carried out using a random effects (DerSimonian-Laird) procedure. Statistically significant associations with melanoma are indicated by a calculated p-value of less than 0.05 for genotype results, which are set forth in bold text.


The absence of a statistically significant association in the replication cohort should not be interpreted as minimizing the value of the original finding. There are many reasons why a biologically derived association identified in a sample from one population would not replicate in a sample from another population. The most important reason is differences in population history. Due to bottlenecks and founder effects, there may be common disease predisposing alleles present in one population that are relatively rare in another, leading to a lack of association in the candidate region. Also, because common diseases such as melanoma are the result of susceptibilities in many genes and many environmental risk factors, differences in population-specific genetic and environmental backgrounds could mask the effects of a biologically relevant allele.


Example 5
NRP1 Proximal SNPs

It has been discovered that a polymorphic variation in a gene encoding the receptor neuropilin 1 (NRP1 is associated with the occurrence of melanoma.


Ninety-five allelic variants located within or nearby the target gene were identified and subsequently allelotyped in melanoma case and control sample sets as described in Examples 1 and 2. The polymorphic variants are set forth in Table 11. The chromosome position provided in column four of Table 11 is based on Genome “Build 34” of NCBI's GenBank and dbSNP 119.

TABLE 11AllelePositionPresentChromo-Chromosomein SEQAllelein SEQIUPACdbSNP rs#somePositionID NO. 1VariantsID NO: 1Coders791040510335900412,141T/CTYrs578433110335908612,961—/AANrs648184510335966838,783T/CTYrs2065364103359800810,108C/TTYrs10643659103359954911,649—/GTTTGNrs7896887103360303115,131A/GGRrs7086456103360641818,518C/TCYrs6481846103360693719,037G/AGRrs7475391103360695519,055C/TTYrs7477281103360695619,056C/TCYrs7475393103360695919,059C/TTYrs7475394103360696619,066C/TTYrs7477282103360697019,070C/TCYrs7475396103360697819,078C/TTYrs2768420103360963821,738G/AGRrs9418057103361248624,586C/TTYrs7070290103361521027,310G/AGRrs1360457103362093533,035T/CARrs1411924103362356835,668G/ACYrs5784335103362617738,277T/—TNrs27769251033718511130,611C/TARrs27769241033718757130,857G/CTSrs19529831033719174131,274A/CTKrs8696361033722278134,378C/TARrs8696371033722444134,544C/TGRrs28044701033723869135,969G/AGRrs27769281033724644136,744T/CTYrs9999661033724924137,024C/TARrs28044751033724926137,026C/GGSrs49348581033728527140,627C/TCYrs49348631033729317141,417C/TTYrs49348641033729327141,427C/TGRrs13313261033730129142,229G/ACYrs13313251033730408142,508A/GTYrs49348711033733328145,428G/AGRrs42770571033736929149,029G/TTKrs27769301033738120150,220C/TARrs13313241033739704151,804G/CGSrs27769321033742645154,745C/TGRrs49348961033743848155,948A/GARrs27769331033745084157,184T/CCYrs49349011033746285158,385A/TTWrs10153001033747483159,583G/CASrs27769341033750081162,181G/CGSrs27769351033750537162,637G/AGRrs27769361033751120163,220C/TTYrs28044911033751126163,226A/CAMrs28044921033752285164,385T/CTYrs28044931033755915168,015A/GGRrs27769371033756428168,528A/GARrs28044941033759655171,755G/CTSrs49349141033761218173,318G/AGRrs27769221033762121174,221T/CCYrs28044951033762739174,839G/TCMrs28044961033764040176,140T/CARrs49345971033764960177,060T/CTYrs28044971033766276178,376G/TAMrs28044981033770946183,046T/CARrs20263191033771741183,841G/CCSrs18886841033773216185,316T/CGRrs37507331033773490185,590A/GCYrs49349271033778301190,401G/CGSrs27769461033782804194,904T/CTYrs49345991033784201196,301T/CTYrs49349341033785140197,240A/GARrs27769441033785311197,411T/GTKrs27769431033786453198,553C/TARrs28048861033787179199,279G/AGRrs28049001033787641199,741G/ATYrs28049021033787672199,772T/CCYrs27683971033788724200,824G/CCSrs27683991033788911201,011A/GARrs27769411033790186202,286C/GASrs28048881033791746203,846G/AARrs28044991033791760203,860C/GTSrs28048891033792103204,203G/AGRrs28048911033792647204,747G/ACYrs27769401033794228206,328A/TAWrs28048971033794781206,881C/TCYrs28048961033795259207,359T/CCYrs27769391033795324207,424C/TCYrs28048941033797330209,430T/CCYrs28045011033798562210,662A/TCWrs21837151033799002211,102A/CTKrs22548221033799071211,171G/ATYrs21506861033799262211,362G/AARrs28048931033799778211,878T/CARrs28045021033800099212,199T/CARrs28048921033800318212,418T/CCYrs21837141033803309215,409G/ATYrs28045031033804955217,055C/TTYrs13204851033806274218,374T/CGRrs49346031033808882220,982A/GTYrs28048901033808922221,022C/TARrs18541451033809260221,360A/GGR


Assay for Verifying and Allelotyping SNPs


The methods used to verify and allelotype the seventy-six proximal SNPs of Table 11 are the same methods described in Examples 1 and 2 herein. The primers used in these assays are provided in Table 12 and Table 13, respectively.

TABLE 12dbSNP rs #Forward PCR PrimerReverse PCR Primerrs6481844GGAGAAAACCAAGATGGCACCATAATTGGCAACTCCCCAGrs7910405GAGTCAGGACTTGCAAACAGCAGACTGAGAACCAACTTAGrs5784331GAATTGGGAGGCATGCTTTGAGCAGATTTACTCCCTTCCCrs6481845AATGCCATCAACTTGGGCTCCAGCAGCTACCAAGCAGTATrs2065364AATCTTGCTGGTGCTCACTCCAGAAGGAGACACTGTCTCArs10643659TTATTACGTAAGTTCCTTCCCAGTCTACCTCTGAAAATAGrs7896887CCTCTCCCACCACTGTTTTAAAGTGCGGTGATAGGAAAGCrs7086456CAGTCTACCTCTGAAAATAGTTATTACGTAAGTTCCTTCCrs6481846ATTGGAATCCCTCCTGGATCTTTTGAACAGGGCACCACAGrs747539TTTAGGAATGGAAATCCTGGGCCTGGTTGTCAGGCTTTAAArs747728TTTATTACGTAAGTTCCTTCCCAGTCTACCTCTGAAAATAGrs7475393AAATAGCTCATGTACCCCAGCCTGCCATAGAGCATATTACrs7475394TTATTACGTAAGTTCCTTCCCAGTCTACCTCTGAAAATAGrs7477282TTATTACGTAAGTTCCTTCCCAGTCTACCTCTGAAAATAGrs7475396TTATTACGTAAGTTCCTTCCCAGTCTACCTCTGAAAATAGrs2768420TTTCCTTGACAGATAACTGCGCAGCTAAAAGACATCATTGrs9418057TTATTACGTAAGTTCCTTCCCAGTCTACCTCTGAAAATAGrs7070290GTGGCACTAGTTTTCAGAAGGCCCTATGAAAACCAGATTCrs1360457CAGGATTATCTGACAGGAGGTATTTTCCTTTCCCACCAGCrs1411924TCCTTCCTCTCCTCCTTCTCCCTACTTTTCCTTCAAACACrs5784335TGATTCACATTCTCCTCCCCTAGGCATGAGCACTTTGTGGrs2776925ACCACTCATGTCTGCCTGCTTGAAGGATCTTTACTCAGGCrs2776924CCCCAAGAATCTTGCACTTCAGGTCTTCAAGACCTGTTCCrs1952983GCTGGCACTCATTTTCTCTCAGTTCCACATAGCTGGAGAGrs869636CACACTCACTGAAGCGTTTGCCAAATAACATGCTGATCGCrs869637AAGTGTTCTTCTCCCACAGCCCTTTAACCCTTACAGAGCCrs2804470GGTGCAGCTAAAGAGAAACGTGGCCGAGGGTTAATTTTTGrs2776928GACAGAAAGATTTGTGACGCCATTTGACAGCTGAGATCCCrs999966GCAGTGGGCACTAACCTAAGGGTTGCGGTTATCTTAGCAGrs2804475TGTGGCATCATTTGTCTTCCCATCATTCCCCCAGTTCTAGrs4934858GATCATTGGTGGAAACAGGGCCACCCATCATTACAATCGCrs4934863GCTTCTATTAGGGTGTTGGCCAGTGCTCACTCAGTTAGTCrs4934864AGTCTCACCAAGACCTTCTGCACCTAGGGATTTATAGACGrs1331326TTTAAACATGGTCTCGTGGCCTACAGTGGCAAAGAACACGrs1331325CCTTCTTCATTCATCAGGGCAAAACTGCCATTCCTCCCACrs4934871GGAATATAAGTCCTGCAGCCTGCACATATAGACGTATCACrs4277057ACACCCTGCATTTTCTCTTGGCAAATGTGCTAAGGCATCGrs2776930CCTCTTCCAAAAGTAAGCCCGGAAGAAACCAGTCCAAGAGrs1331324GTGCTTATTACCCGAGTGACCCTGTACGTGTACAGGTTTGrs2776932CTGCAAGAACCTGAACTCTGAGAAATCGAGGTGTTGGCTGrs4934896ACAGATATGCTGCAGTGACCATTACCCAAGTAGGCTTCCCrs2776933CTGAGTATCTGGGACTACAGTGGTTAACACGGTGAAACCCrs4934901GCACTAGTCCAGCATTTTCCACTTTGCTTCATACCATGGArs1015300CCTCTTCTGTAATCTCTAGTGAACGGACAAGTACTCTAAGCrs2776934CTCCCTTTCTTGAGGTTGACCAAACACGAATCTAGGAGCCrs2776935AATACTCTCACCACCACCACCCCAGCACAAATAAGTGCTCrs2776936CAATCCCAGGGGATTATAGCTGTTTACCGATGCTCGTACCrs2804491AACTCCTGACCTCAAGTGACAAATGAACTCCTCTTGGCCGrs2804492ATGCCTGAAATCCCAGCTACAGTGGCTCAATCTTGGCTCCrs2804493TCAGCTAATTCCCAGCATTCGGAGTTGACAGTTCAAATCCTrs2776937GAAATCCTGTGGTAGGAATCGACATCTCATTTTCTAGGTGrs2804494CCTAGACCTTAGTCTTACCCTCAGAAGACCACATTCTCAGrs4934914TAAGTCCTGCAGCCACTTTGGCATACACAAATATATGCACrs2776922TTGAAATCCTTTCCACTGGCGTTCTGTTGCCTTCATGCTCrs2804495GGAAGATCTTTGAGAGTTAGCAGGAACAGCCACATTTTGACrs2804496AAAGAAGAAGGCCCCAACACTTCTTGGCCCTGATTTCTCCrs4934597TTCTTGACCTCGTGATCCACAATTAACCCTCGGCCAGGTGrs2804497AACTCCCTCCTTCCCTTAACCTCAGGGTGTATTTTACACGrs2804498AGCACTTAAGACAGCCACTCGTGGCCAGATTCTCTGAATGrs2026319AATTGGCATGCTGATCTGGGGGGCTTTTCCTTACCTCTGTrs1888684AACAGCCTTTCAGCTTTGGCTTGTCCCAACTGAGGCTTTGrs3750733CTGCCTGTCACTTACCGTTGGCTCCTCTGCGCCGTGCTCrs4934927TTTATTCATGGCAGAAGGCGATAAAGTGCAGTACCTCCCCrs2776946CCCATAAAGGAAGCAGAAGCTTTGTCCCCTATCCTCTCTCrs4934599TTTTCGAGACGGAGTCTTGCGAGGCGGAGATTACAGTAAGrs4934934GGATGAAATGAGAATGACCCGAGGAGTCGACTGTTGAATGrs2776944CATAAGGCCATTCATGAGGGGAAAGAGCTTGGGATCATGGrs2776943GGAGTTGACAGTTCAAATCCTTCAGCTAATTCCCAGCATTCrs2804886GAAAAGAACATTATTTGCAGCCTCCCTTTTTCTTATTAGTCrs2804900CTAATTTCACCTGTAGGATGTTCACATGGTGAAAGCAGGGrs2804902AAGTTTGAGAACCACTGGGCCATGAATCCCTCTGCAACTCrs2768397CACACCAGAGCTCTTTAATCGAGAGATTGAACAGAGCCAGrs2768399CAAGATGAGATTTGGGTGGGTGGACTGGTTTCTTCCTCACrs277694TTGGTTTATGGTCTTGGCGTCGGGAAGAAAAGACGACCTACrs2804888TAACATGTCCACAGCCTTCCGGATGGTTTCACTACTGTATGrs2804499TGGCTCACTCGCAAACATTGATGTTGGCCAGGCTGGTTTCrs2804889ACACCCAACTCTCTATGAGCAGGTCCTGACCATGCATGAGrs2804891GTGAACAAAAGTAGACTCCCGCACTCTCTGGGATAATTTCrs2776940TACATGAAGGCAATCGCACCTGGTGGTGGATAAGCCAATGrs2804897ATCTGGTCAGGTTTGGTCTCCAAATAGAACTGACATCTGCrs2804896AAGACCCTCTTCCTGGTTTGTGCTTTCTCTCTCTTCTCCCrs2776939TTTCCAGCTTGCAGAACCTCTAGAGCATGATCTAGGTCCCrs2804894CATTAATATGTGGCCATGGGAGGGGTACACAATTCAATCCrs2804501AGTTCGTTGCACCTTGAGAGGACACTACTAACTGCCCTAGrs2183715ATGGATTGAGCCACACTCTCTGTCTCTAGACGTGTGTTGGrs2254822TTCAGTGAAAGTTGCCTGTGAGAGGTGCAATAACTGGTGGrs2150686TTCAGTGAAAGTTGCCTGTGAGAGGTGCAATAACTGGTGGrs2804893ATACAAAAATTAGCCGGGCGCTCCTGGGTTCAAGAGATTCrs2804502AATGTGGCAGCATTTCAGGCCTGGGCTGCCTATATTGTTCrs2804892TAATTTCCTCCATCCCTCCCCCCATGAAAGACTTAACCTTCrs2183714TTTAAACATGGTCTCGTGGCCTACAGTGGCAAAGAACACGrs2804503GTCCTCTCTCTGTTTTAGGCAATCTGTTTGCTTACTTTTCrs1320485ATGCCTGAAATCCCAGCTACAGTGGCTCAATCTTGGCTCCrs4934603CTCTCCTCTGTGTAACCAACGTTACAAGTATGCGATCTTCCrs2804890ACTTAAGTGGCATCAGGTAGATCACAAATTGGAAGCCAACrs1854145GGATCCTAATTTGCGCTGACTCTGGGCAGTAAGTTTCAGG












TABLE 13













Term












dbSNP rs #
Extend Primer
Mix
















rs6481844
AACAACTGGAAAACCTTGGCT
ACT








rs7910405
GATGTTCAGTGCTTGCTTTGC
ACT







rs5784331
ATGATTCCAGGGGATTTTTTTTT
ACG







rs6481845
AGAAGGTGAATGCTATAACTAAA
ACT







rs2065364
TCTCCTCCCCATGTGCT
ACG







rs10643659
CATATTATTTCAGAAATTATGGTTG
ACT







rs7896887
CTGGGACTACAGACGCCC
ACT







rs7086456
TCATGTACCCCAGTAATGTATA
ACG







rs6481846
GAGAGAAGGAAGGAAGGAAAA
ACG







rs7475391
TTCGTTTTCCTTCCTTCCTTC
ACG







rs7477281
TCGTTTTCCTTCCTTCCTTCT
ACG







rs7475393
TTTTCCTTCCTTCCTTCTCTC
ACG







rs7475394
CCTTCCTTCTCTCTCTCTCT
ACG







rs7477282
TCCTTCTCTCTCTCTCTTTCT
ACG







rs7475396
TCTCTCTCTTTCTCTCTCTCT
ACG







rs2768420
CACCATGAGATGCAGGA
ACG







rs9418057
TGGTGCTCACTCTTTGCGG
ACG







rs7070290
GATGGAGTTTAAAAGCACAAAG
ACG







rs1360457
CAGCTTTGGCCAGGAGATG
ACT







rs1411924
ATTTATAATCTACATTGTACACAC
ACG







rs5784335
TTCCATGTTCATAATTTTTTTTTT
ACT







rs2776925
CCAGTGGTGTGAGCTTC
ACG







rs2776924
GAGGTTGACTCATGATTT
ACT







rs1952983
CCTGATTGAAGCAAAACCAGAA
ACT







rs869636
TGTTCAACAGACCCTTC
ACG







rs869637
GTAATCTCTAGTGTTTATTGAAAG
ACG







rs2804470
TTGCAGCATTTCCCTAA
ACG







rs2776928
GTGGATCCTAATGATCAC
ACT







rs999966
GGAGAACTGCTTGAACC
ACG







rs2804475
AGAACTGCTTGAACCCG
ACT







rs4934858
TGGGATTTCGTTACCTAA
ACG







rs4934863
CAACCTGAGGATGTATAGAA
ACG







rs4934864
GGATGTATAGAATGCCTATATG
ACG







rs1331326
TTCCTTGATCTCTTTCTTC
ACG







rs1331325
TTGCGCTGACAAAAACC
ACT







rs4934871
GAGATATGTAGGACCCC
ACG







rs4277057
AGAGGGTAGTAAATCCTG
CGT







rs2776930
TGCTATTACTACTTCTACCTAC
ACG







rs1331324
GTGTGTGCACATCTCAA
ACT







rs2776932
CGTAGTTCTTCGCCCCT
ACG







rs4934896
ACCCCATCTAAACAAAAG
ACT







rs2776933
TAGCCGGGCGTGGTGGT
ACT







rs4934901
TGGGATCATGAATCCTAG
CGT







rs1015300
CTAAACCCTTGTGACATG
ACT







rs2776934
TTTTACTGACTCTTCAAGTG
ACT







rs2776935
TATCTTCATCTGCAGCC
ACG







rs2776936
GACAGTTCAAATCCTTTTTTT
ACG







rs280449
TTGTCTAGTATTTCTAAACACTG
ACT







rs2804492
CTTAGTCTTACCCATGAAC
ACT







rs2804493
GGAGTTGCCAATTATGTATAAG
ACT







rs2776937
GGTGGGGATAAGGAGCT
ACT







rs2804494
GGGTGTTGGAGTGGGTG
ACT







rs4934914
CCCTCATTCATACAGTTT
ACG







rs2776922
TTTGTGACGCAGCAACT
ACT







rs2804495
GGTTGATTGGCCTGTAT
CGT







rs2804496
AGGCAGGCATCACTTGA
ACT







rs4934597
TTTTTCCTAACTCAGTGC
ACT







rs2804497
GTATAGTCAACTTCCACC
CGT







rs2804498
TCCATCATAAAATGTGACAAT
ACT







rs2026319
TGACTTCTGAGAAAGTGA
ACT







rs1888684
ATGCTGATCTGGGAAGGTAG
ACT







rs3750733
TTACCGTTGCGAAAAGC
ACT







rs4934927
TGAATCTTTGGTAACACC
ACT







rs2776946
GTGCTAGGATTACAGGC
ACT







rs4934599
CACAGAGCTCACCCACC
ACT







rs4934934
AGGAAAAAGGCTTATCAAG
ACT







rs2776944
ACTTGACCTAGCACCAG
ACT







rs2776943
GGCTAAAAAGTGTGTCAA
ACG







rs2804886
GATGTGCTCAGCTCTTA
ACG







rs2804900
CTGTAATCTCCGCCTCC
ACG







rs2804902
GGCTGGAGTGCAGTGGC
ACT







rs2768397
CTGGCTAGCCTTTTAGAA
ACT







rs2768399
ACAAAAACCACCATCAAA
ACT







rs2776941
ATGCCAACTGAGAAGGA
ACT







rs2804888
CTCCCTTCTGGGAGGAT
ACG







rs2804499
CCCAATCCTCCCAGAAG
ACT







rs2804889
GGTCTCCAAATGAGTGG
ACG







rs2804891
GAAACAGGGATGGAGATA
ACG







rs2776940
CAGCAACAGGTCTCACC
CGT







rs2804897
GAGAGGAATGGAGAAAAA
ACG







rs2804896
GTACTGGGATTACAGGC
ACT







rs2776939
AAACGTCAGAGCCAGCC
ACG







rs2804894
TCTCACTGGGGACCCAC
ACT







rs2804501
GCATTTTCCTCTCTCTG
CGT







rs2183715
GATAGAGGTCAAGGAGAG
ACT







rs2254822
GTAAGCCCTACCTATGC
ACG







rs2150686
GTTGCCTGTGTTCTGGA
ACG







rs2804893
GCAATTCTCCTGCCTCA
ACT







rs2804502
GTGAATATGGAAAAAACCAA
ACT







rs2804892
TGCCATCTTCTCATTGT
ACT







rs2183714
TTGTGTAGAATAGAGCCC
ACG







rs2804503
TTGTCCTCTGTGCTAGA
ACG







rs1320485
TGCTGGTGAGCAGGGAG
ACT







rs4934603
GCAGAAGGCGAAGTGGT
ACT







rs2804890
GTGCAGTACCTCCCCCC
ACG







rs1854145
CTCATTTTCTCTCTATTCCC
ACT










Genetic Analysis


Allelotyping results are shown for female (F) and male (M) cases and controls in Table 14 and Table 15, respectively. Allele frequency is noted in the fourth and fifth columns for melanoma pools and control pools, respectively.

TABLE 14FemalesMelanomaChromo-ChromosomeFemale CaseFemale ControlFOddsAssociateddbSNP rs#somePositionAllelesAFAFp-ValueRatioAllelers64818441033588137T/C T = 0.587 T = 0.6120.4401.110C  C = 0.412   C = 0.387 rs79104051033590041T/C T = 0.926 T = 0.8900.0900.640T  C = 0.073   C = 0.109 rs57843311033590861—/A— = 0.375  — = 0.386  0.7251.049AA = 0.624A = 0.613rs64818451033596683T/C T = 0.329 T = 0.3050.4450.896T  C = 0.670   C = 0.694 rs20653641033598008A/GA = 0.354A = 0.3960.1921.197GG = 0.645G = 0.603rs106436591033599549—/GTTT— = 0.198  — = 0.125  0.0060.582GTTT = 0.801    GTTT = 0.874    rs78968871033603031A/GA = 0.631A = 0.5710.0660.778AG = 0.368G = 0.428rs70864561033606418C/T  C = 0.857   C = 0.853 0.8700.965C T = 0.142 T = 0.146rs64818461033606937G/AG = 0.849G = 0.8370.6630.919GA = 0.150A = 0.162rs74753911033606955C/T  C = 0.705   C = 0.608 0.0140.647C T = 0.294 T = 0.391rs74772811033606956C/T  C = 0.291   C = 0.236 0.0790.752C T = 0.708 T = 0.763rs74753931033606959C/T  C = 0.549   C = 0.486 0.1590.777C T = 0.450 T = 0.513rs74753941033606966C/T  C = 0.729   C = 0.815 0.0071.641T T = 0.270 T = 0.184rs74772821033606970C/T  C = 0.908   C = 0.899 0.7380.903C T = 0.091 T = 0.100rs74753961033606978C/T  C = 0.209   C = 0.104 ˜0.0010.438C T = 0.790 T = 0.895rs27684201033609638T/C T = 0.789 T = 0.7910.9251.015C  C = 0.210   C = 0.208 rs94180571033612486C/T  C = 0.865   C = 0.916 0.0191.698T T = 0.134 T = 0.083rs70702901033615210G/AG = 0.948G = 0.9410.7250.879GA = 0.051A = 0.058rs13604571033620935T/C T = 0.278 T = 0.2100.0200.692T  C = 0.721   C = 0.789 rs14119241033623568G/AG = 0.165G = 0.1260.1100.729GA = 0.834A = 0.873rs57843351033626177T/— T = 0.219 T = 0.129˜0.0010.526T— = 0.780  — = 0.870  rs27769251033572278C/T  C = 0.436   C = 0.452 0.6081.071T T = 0.563 T = 0.547rs27769241033572524G/CG = 0.755G = 0.7240.3070.850G  C = 0.244   C = 0.275 rs19529831033572941A/CA = 0.616A = 0.5530.0810.771A  C = 0.383   C = 0.446 rs8696361033576045C/T  C = 0.442   C = 0.444 0.9491.009T T = 0.557 T = 0.555rs8696371033576211C/T  C = 0.888   C = 0.914 0.2251.338T T = 0.111 T = 0.085rs28044701033577636G/AG = 0.181G = 0.2400.0711.422AA = 0.818A = 0.759rs27769281033578411T/C T = 0.893 T = 0.9200.1771.385C  C = 0.106   C = 0.079 rs9999661033578691C/T  C = 0.828   C = 0.828 0.9951.001T T = 0.171 T = 0.171rs28044751033578693C/G  C = 0.035   C = 0.061 0.1771.808GG = 0.964G = 0.938rs49348581033582294C/T  C = 0.505   C = 0.525 0.5521.084T T = 0.494 T = 0.474rs49348631033583084C/T  C = 0.190   C = 0.189 0.9660.992C T = 0.809 T = 0.810rs49348641033583094C/T  C = 0.536   C = 0.508 0.4260.892C T = 0.463 T = 0.491rs13313261033583896G/AG = 0.437G = 0.4650.4361.118AA = 0.562A = 0.534rs13313251033584175A/GA = 0.261A = 0.1890.0090.660AG = 0.738G = 0.810rs49348711033587095G/AG = 0.769G = 0.7090.0390.734GA = 0.230A = 0.290rs42770571033590696G/TG = 0.867G = 0.8600.7470.938G T = 0.132 T = 0.139rs27769301033591887C/T  C = 0.210   C = 0.159 0.0480.708C T = 0.789 T = 0.840rs13313241033593471G/CG = 0.621G = 0.6110.7640.960G  C = 0.378   C = 0.388 rs27769321033596412C/T  C = 0.189   C = 0.227 0.1721.265T T = 0.810 T = 0.772rs49348961033597615A/GA = 0.860A = 0.8290.2190.790AG = 0.139G = 0.170rs27769331033598851T/CT =    T = 0.535C =      C = 0.465 rs49349011033600052A/TA = 0.837A = 0.8040.1990.800A T = 0.162 T = 0.195rs10153001033601250G/CG = 0.948G = 0.9560.7131.200C  C = 0.051   C = 0.043 rs27769341033603848G/CG = 0.256G = 0.2930.2221.204C  C = 0.743   C = 0.706 rs27769351033604304G/AG = 0.459G = 0.3790.0130.719GA = 0.540A = 0.620rs27769361033604887C/T  C = 0.293   C = 0.213 0.0060.654C T = 0.706 T = 0.786rs28044911033604893A/CA = 0.236A = 0.1770.0350.700A  C = 0.763   C = 0.822 rs28044921033606052T/C T = 0.603 T = 0.6270.4921.108C  C = 0.396   C = 0.372 rs28044931033609682A/GA = 0.234A = 0.2100.3830.868AG = 0.765G = 0.789rs27769371033610195A/GA = 0.642A = 0.5990.2690.832AG = 0.357G = 0.400rs28044941033613422G/CG = 0.381G = 0.3110.0440.734G  C = 0.618   C = 0.688 rs49349141033614985G/AG = 0.920G = 0.9260.7201.098AA = 0.079A = 0.073rs27769221033615888T/C T = 0.293 T = 0.2050.0030.622T  C = 0.706   C = 0.794 rs28044951033616506G/TG = 0.399G = 0.3230.0310.717G T = 0.600 T = 0.676rs28044961033617807T/C T = 0.364 T = 0.250˜0.0010.582T  C = 0.635   C = 0.749 rs49345971033618727T/C T = 0.224 T = 0.1970.3340.849T  C = 0.775   C = 0.802 rs28044971033620043G/TG = 0.968G = 0.9700.8911.068T T = 0.031 T = 0.029rs28044981033624713T/C T = 0.757 T = 0.7700.6441.077C  C = 0.242   C = 0.229 rs20263191033625508G/CG = 0.901G = 0.9080.7421.083C  C = 0.098   C = 0.091 rs18886841033626983T/C T = 0.295 T = 0.2280.0510.707T  C = 0.704   C = 0.771 rs37507331033627257A/GA = 0.177A =     G = 0.823G =     rs49349271033632068G/CG = 0.896G = 0.9060.6081.127C  C = 0.103   C = 0.093 rs27769461033636571T/C T = 0.375 T = 0.3290.1900.818T  C = 0.624   C = 0.670 rs49345991033637968T/C T = 0.088 T = 0.0740.4790.834T  C = 0.911   C = 0.925 rs49349341033638907A/GA = 0.917A = 0.9150.9110.973AG = 0.082G = 0.084rs27769441033639078T/G T = 0.550 T = 0.5200.4480.887TG = 0.449G = 0.479rs27769431033640220C/T  C = 0.314   C = 0.259 0.0770.765C T = 0.685 T = 0.740rs28048861033640946G/AG = 0.870G = 0.9090.0771.495AA = 0.129A = 0.090rs28049001033641408G/AG = 0.754G = 0.7390.6440.925GA = 0.246A = 0.261rs28049021033641439T/C T = 0.692 T = 0.6800.6940.945T  C = 0.307   C = 0.319 rs27683971033642491G/CG = 0.904G = 0.9340.1871.515C  C = 0.095   C = 0.065 rs27683991033642678A/GA = 0.245A = 0.2020.1720.782AG = 0.754G = 0.797rs27769411033643953C/G  C = 0.697   C = 0.731 0.2651.185GG = 0.302G = 0.268rs28048881033645513G/AG = 0.053G = 0.0720.2901.392AA = 0.946A = 0.927rs28044991033645527C/G  C = 0.796   C = 0.835 0.1841.298GG = 0.203G = 0.164rs28048891033645870G/AG = 0.815G = 0.8590.0831.380AA = 0.184A = 0.140rs28048911033646414G/AG = 0.303G = 0.2320.0150.696GA = 0.696A = 0.767rs27769401033647995A/TA = 0.274A = 0.1970.0090.654A T = 0.725 T = 0.802rs28048971033648548C/T  C = 0.842   C = 0.900 0.0111.691T T = 0.157 T = 0.099rs28048961033649026T/C T = 0.540 T = 0.4950.1930.835T  C = 0.459   C = 0.504 rs27769391033649091C/T  C = 0.400   C = 0.359 0.2140.841C T = 0.599 T = 0.640rs28048941033651097T/C T = 0.747 T = 0.7970.0781.328C  C = 0.252   C = 0.202 rs28045011033652329A/TA = 0.381A = 0.3730.8170.969A T = 0.618 T = 0.626rs21837151033652769A/CA = 0.904A = 0.9070.8841.037C  C = 0.095   C = 0.092 rs22548221033652838G/AG =     G =     A =     A =     rs21506861033653029G/AG = 0.116G = 0.1260.6321.105AA = 0.883A = 0.873rs28048931033653545T/C T = 0.173 T = 0.1320.1440.729T  C = 0.827   C = 0.868 rs28045021033653866T/C T = 0.765 T = 0.8140.0701.347C  C = 0.234   C = 0.185 rs28048921033654085T/C T = 0.871 T = 0.9110.0631.515C  C = 0.128   C = 0.088 rs21837141033657076G/AG = 0.267G = 0.1850.0120.627GA = 0.732A = 0.814rs28045031033658722C/T  C = 0.416   C = 0.376 0.2370.845C T = 0.583 T = 0.623rs13204851033660041T/C T = 0.403 T = 0.3920.7450.956T  C = 0.596   C = 0.607 rs49346031033662649A/GA = 0.585A = 0.6420.1121.272GG = 0.414G = 0.357rs28048901033662689C/TC =      C = 0.559 T =    T = 0.441rs18541451033663027A/GA = 0.042A = 0.0290.4850.674AG = 0.957G = 0.970









TABLE 15










Males























Melanoma



Chromo-
Chromosome

Male Case
Male Control
M
Odds
Associated


dbSNP rs#
some
Position
Alleles
AF
AF
p-Value
Ratio
Allele





rs6481844
10
33588137
T/C
 T = 0.625
 T = 0.575
0.134
0.810
T







  C = 0.374 


  C = 0.424 



rs7910405
10
33590041
T/C
 T = 0.906
 T = 0.872
0.129
0.703
T







  C = 0.093 


  C = 0.127 



rs5784331
10
33590861
—/A
— = 0.386  
— = 0.386  
0.998
1.000
A






A = 0.613
A = 0.613


rs6481845
10
33596683
T/C
 T = 0.342
 T = 0.252

0.004

0.647
T







  C = 0.657 


  C = 0.747 



rs2065364
10
33598008
A/G
A = 0.397
A = 0.338
0.086
0.775
A






G = 0.602
G = 0.661


rs10643659
10
33599549
—/GTTT
— = 0.185  
— = 0.123  

0.022

0.617







GTTT = 0.814    
GTTT = 0.876    


rs7896887
10
33603031
A/G
A = 0.600
A = 0.560
0.256
0.845
A






G = 0.399
G = 0.439


rs7086456
10
33606418
C/T

  C = 0.838 


  C = 0.866 

0.321
1.247
T






 T = 0.161
 T = 0.133


rs6481846
10
33606937
G/A
G = 0.810
G = 0.863
0.059
1.474
A






A = 0.189
A = 0.136


rs7475391
10
33606955
C/T

  C = 0.675 


  C = 0.466 


˜0.001

0.419
C






 T = 0.324
 T = 0.533


rs7477281
10
33606956
C/T

  C = 0.271 


  C = 0.238 

0.300
0.840
C






 T = 0.728
 T = 0.761


rs7475393
10
33606959
C/T

  C = 0.502 


  C = 0.463 

0.333
0.856
C






 T = 0.497
 T = 0.536


rs7475394
10
33606966
C/T

  C = 0.757 


  C = 0.835 


0.006

1.630
T






 T = 0.242
 T = 0.164


rs7477282
10
33606970
C/T
C =    
C =    






T =   
T =   


rs7475396
10
33606978
C/T
C =    
C =    






T =   
T =   


rs2768420
10
33609638
T/C
 T = 0.780
 T = 0.782
0.941
1.013
C







  C = 0.219 


  C = 0.217 



rs9418057
10
33612486
C/T

  C = 0.870 


  C = 0.917 


0.035

1.654
T






 T = 0.129
 T = 0.082


rs7070290
10
33615210
G/A
G = 0.941
G = 0.952
0.557
1.227
A






A = 0.058
A = 0.047


rs1360457
10
33620935
T/C
 T = 0.264
 T = 0.204
0.081
0.716
T







  C = 0.735 


  C = 0.795 



rs1411924
10
33623568
G/A
G = 0.134
G = 0.098
0.121
0.701
G






A = 0.865
A = 0.901


rs5784335
10
33626177
T/—
 T = 0.183
 T = 0.113

0.024

0.574
T






— = 0.816  
— = 0.886  


rs2776925
10
33572278
C/T

  C = 0.440 


  C = 0.459 

0.571
1.082
T






 T = 0.559
 T = 0.540


rs2776924
10
33572524
G/C
G = 0.751
G = 0.726
0.470
0.881
G







  C = 0.248 


  C = 0.273 



rs1952983
10
33572941
A/C
A = 0.613
A = 0.590
0.511
0.908
A







  C = 0.386 


  C = 0.409 



rs869636
10
33576045
C/T

  C = 0.428 


  C = 0.440 

0.720
1.053
T






 T = 0.571
 T = 0.559


rs869637
10
33576211
C/T

  C = 0.944 


  C = 0.941 

0.908
0.958
C






 T = 0.055
 T = 0.058


rs2804470
10
33577636
G/A
G = 0.189
G = 0.167
0.415
0.858
G






A = 0.810
A = 0.832


rs2776928
10
33578411
T/C
 T = 0.918
 T = 0.936
0.361
1.295
C







  C = 0.081 


  C = 0.063 



rs999966
10
33578691
C/T

  C = 0.822 


  C = 0.827 

0.840
1.039
T






 T = 0.177
 T = 0.172


rs2804475
10
33578693
C/G

  C = 0.052 


  C = 0.032 

0.435
0.598
C






G = 0.947
G = 0.967


rs4934858
10
33582294
C/T
C =    
C =    






T =   
T =   


rs4934863
10
33583084
C/T

  C = 0.203 


  C = 0.236 

0.294
1.207
T






 T = 0.796
 T = 0.763


rs4934864
10
33583094
C/T

  C = 0.572 


  C = 0.520 

0.163
0.810
C






 T = 0.427
 T = 0.479


rs1331326
10
33583896
G/A
G = 0.441
G = 0.420
0.586
0.916
G






A = 0.558
A = 0.579


rs1331325
10
33584175
A/G
A = 0.205
A = 0.149

0.037

0.678
A






G = 0.794
G = 0.850


rs4934871
10
33587095
G/A
G = 0.751
G = 0.754
0.921
1.018
A






A = 0.248
A = 0.245


rs4277057
10
33590696
G/T
G = 0.843
G = 0.855
0.626
1.105
T






 T = 0.156
 T = 0.144


rs2776930
10
33591887
C/T

  C = 0.226 


  C = 0.170 

0.056
0.702
C






 T = 0.773
 T = 0.829


rs1331324
10
33593471
G/C
G = 0.631
G = 0.572
0.086
0.782
G







  C = 0.368 


  C = 0.427 



rs2776932
10
33596412
C/T

  C = 0.206 


  C = 0.175 

0.328
0.816
C






 T = 0.793
 T = 0.824


rs4934896
10
33597615
A/G
A = 0.841
A = 0.826
0.582
0.896
A






G = 0.158
G = 0.173


rs2776933
10
33598851
T/C
T =   
 T = 0.495






C =    

  C = 0.505 



rs4934901
10
33600052
A/T
A = 0.812
A = 0.826
0.689
1.104
T






 T = 0.187
 T = 0.173


rs1015300
10
33601250
G/C
G = 0.966
G = 0.936
0.124
0.519
G







  C = 0.033 


  C = 0.063 



rs2776934
10
33603848
G/C
G = 0.256
G = 0.219
0.215
0.816
G







  C = 0.743 


  C = 0.780 



rs2776935
10
33604304
G/A
G = 0.482
G = 0.387

0.006

0.680
G






A = 0.517
A = 0.612


rs2776936
10
33604887
C/T
C =    
C =    






T =   
T =   


rs2804491
10
33604893
A/C
A = 0.225
A = 0.167

0.047

0.694
A







  C = 0.774 


  C = 0.832 



rs2804492
10
33606052
T/C
 T = 0.635
 T = 0.587
0.186
0.817
T







  C = 0.364 


  C = 0.412 



rs2804493
10
33609682
A/G
A = 0.212
A = 0.210
0.935
0.986
A






G = 0.787
G = 0.789


rs2776937
10
33610195
A/G
A = 0.621
A = 0.582
0.303
0.849
A






G = 0.378
G = 0.417


rs2804494
10
33613422
G/C
G = 0.363
G = 0.314
0.138
0.802
G







  C = 0.636 


  C = 0.685 



rs4934914
10
33614985
G/A
G = 0.930
G = 0.949
0.279
1.405
A






A = 0.069
A = 0.050


rs2776922
10
33615888
T/C
 T = 0.265
 T = 0.223
0.164
0.796
T







  C = 0.734 


  C = 0.776 



rs2804495
10
33616506
G/T
G = 0.350
G = 0.322
0.392
0.883
G






 T = 0.649
 T = 0.677


rs2804496
10
33617807
T/C
 T = 0.332
 T = 0.212

0.001

0.542
T







  C = 0.667 


  C = 0.787 



rs4934597
10
33618727
T/C
 T = 0.192
 T = 0.160
0.235
0.801
T







  C = 0.807 


  C = 0.839 



rs2804497
10
33620043
G/T
G = 0.977
G = 0.995
0.108
4.348
T






 T = 0.023
 T = 0.005


rs2804498
10
33624713
T/C
 T = 0.768
 T = 0.777
0.759
1.053
C







  C = 0.231 


  C = 0.222 



rs2026319
10
33625508
G/C
G = 0.919
G = 0.949
0.310
1.636
C







  C = 0.080 


  C = 0.050 



rs1888684
10
33626983
T/C
 T = 0.245
 T = 0.161

0.038

0.590
T







  C = 0.754 


  C = 0.838 



rs3750733
10
33627257
A/G
A =     
A =     






G =     
G =     


rs4934927
10
33632068
G/C
G = 0.908
G = 0.934
0.193
1.455
C







  C = 0.091 


  C = 0.065 



rs2776946
10
33636571
T/C
 T = 0.351
 T = 0.315
0.326
0.853
T







  C = 0.648 


  C = 0.684 



rs4934599
10
33637968
T/C
 T = 0.084
 T = 0.060
0.226
0.707
T







  C = 0.915 


  C = 0.939 



rs4934934
10
33638907
A/G
A = 0.923
A = 0.935
0.552
1.191
G






G = 0.076
G = 0.064


rs2776944
10
33639078
T/G
T =   
T =   






G =     
G =     


rs2776943
10
33640220
C/T

  C = 0.266 


  C = 0.235 

0.307
0.849
C






 T = 0.733
 T = 0.764


rs2804886
10
33640946
G/A
G = 0.905
G = 0.934
0.179
1.485
A






A = 0.094
A = 0.065


rs2804900
10
33641408
G/A
G = 0.746
G = 0.738
0.810
0.957
G






A = 0.254
A = 0.262


rs2804902
10
33641439
T/C
 T = 0.710
 T = 0.678
0.327
0.860
T







  C = 0.289 


  C = 0.321 



rs2768397
10
33642491
G/C
G = 0.951
G = 0.945
0.753
0.892
G







  C = 0.048 


  C = 0.054 



rs2768399
10
33642678
A/G
A = 0.200
A = 0.154
0.108
0.727
A






G = 0.799
G = 0.845


rs2776941
10
33643953
C/G

  C = 0.739 


  C = 0.731 

0.836
0.963
C






G = 0.260
G = 0.268


rs2804888
10
33645513
G/A
G = 0.034
G = 0.074

0.043

2.246
A






A = 0.965
A = 0.925


rs2804499
10
33645527
C/G

  C = 0.855 


  C = 0.856 

0.993
1.002
G






G = 0.144
G = 0.143


rs2804889
10
33645870
G/A
G = 0.854
G = 0.879
0.323
1.238
A






A = 0.145
A = 0.120


rs2804891
10
33646414
G/A
G = 0.282
G = 0.240
0.167
0.805
G






A = 0.717
A = 0.759


rs2776940
10
33647995
A/T
A = 0.235
A = 0.196
0.170
0.793
A






 T = 0.764
 T = 0.803


rs2804897
10
33648548
C/T

  C = 0.892 


  C = 0.924 

0.170
1.467
T






 T = 0.107
 T = 0.075


rs2804896
10
33649026
T/C
 T = 0.453
 T = 0.525
0.059
1.331
C







  C = 0.546 


  C = 0.474 



rs2776939
10
33649091
C/T

  C = 0.370 


  C = 0.349 

0.535
0.913
C






 T = 0.629
 T = 0.650


rs2804894
10
33651097
T/C
 T = 0.779
 T = 0.816
0.206
1.257
C







  C = 0.220 


  C = 0.183 



rs2804501
10
33652329
A/T
A = 0.357
A = 0.374
0.625
1.072
T






 T = 0.642
 T = 0.626


rs2183715
10
33652769
A/C
A = 0.921
A = 0.925
0.856
1.051
C







  C = 0.078 


  C = 0.074 



rs2254822
10
33652838
G/A
G = 0.420
G = 0.419
0.979
0.996
G






A = 0.579
A = 0.580


rs2150686
10
33653029
G/A
G = 0.106
G = 0.112
0.800
1.060
A






A = 0.893
A = 0.887


rs2804893
10
33653545
T/C
 T = 0.145
 T = 0.127
0.515
0.857
T







  C = 0.855 


  C = 0.873 



rs2804502
10
33653866
T/C
 T = 0.799
 T = 0.810
0.689
1.077
C







  C = 0.200 


  C = 0.189 



rs2804892
10
33654085
T/C
 T = 0.903
 T = 0.932
0.174
1.465
C







  C = 0.096 


  C = 0.067 



rs2183714
10
33657076
G/A
G = 0.219
G = 0.187
0.300
0.825
G






A = 0.780
A = 0.812


rs2804503
10
33658722
C/T

  C = 0.385 


  C = 0.380 

0.871
0.977
C






 T = 0.614
 T = 0.619


rs1320485
10
33660041
T/C
 T = 0.411
 T = 0.352
0.124
0.776
T







  C = 0.588 


  C = 0.647 



rs4934603
10
33662649
A/G
A = 0.591
A = 0.620
0.420
1.131
G






G = 0.408
G = 0.379


rs2804890
10
33662689
C/T

  C = 0.476 

C =    






 T = 0.524
T =   


rs1854145
10
33663027
A/G
A = 0.019
A = 0.067

0.007

3.680
G






G = 0.981
G = 0.933









Allelotyping results were considered particularly significant with a calculated p-value of less than or equal to 0.05 for allelotype results. These values are indicated in bold. The assay failed for those SNPs in which the allele frequency is blank. The combined allelotyping p-values for males and females were plotted in FIG. 1A and separately for females and males in FIGS. 1B and 1C, respectively. The position of each SNP on the chromosome is presented on the x-axis. The y-axis gives the negative logarithm (base 10) of the p-value comparing the estimated allele frequency in the case group to that of the control group. The minor allele frequency of the control group for each SNP designated by an X or other symbol on the graphs in FIGS. 1A, 1B and 1C can be determined by consulting Table 14 or 15. By proceeding down the Table from top to bottom and across the graphs from left to right the allele frequency associated with each symbol shown can be determined.


To aid the interpretation, multiple lines have been added to the graph. The broken horizontal lines are drawn at two common significance levels, 0.05 and 0.01. The vertical broken lines are drawn every 20 kb to assist in the interpretation of distances between SNPs. Two other lines are drawn to expose linear trends in the association of SNPs to the disease. The light gray line (or generally bottom-most curve) is a nonlinear smoother through the data points on the graph using a local polynomial regression method (W. S. Cleveland, E. Grosse and W. M. Shyu (1992) Local regression models. Chapter 8 of Statistical Models in S eds J. M. Chambers and T. J. Hastie, Wadsworth & Brooks/Cole.). The black line (or generally top-most curve, e.g., see peak in left-most graph just to the left of position 92150000) provides a local test for excess statistical significance to identify regions of association. This was created by use of a 10 kb sliding window with 1 kb step sizes. Within each window, a chi-square goodness of fit test was applied to compare the proportion of SNPs that were significant at a test wise level of 0.01, to the proportion that would be expected by chance alone (0.05 for the methods used here). Resulting p-values that were less than 1-8 were truncated at that value.


Finally, the exons and introns of the genes in the covered region are plotted below each graph at the appropriate chromosomal positions. The gene boundary is indicated by the broken horizontal line. The exon positions are shown as thick, unbroken bars. An arrow is placed at the 3′ end of each gene to show the direction of transcription.


Example 6
ENDO180 Proximal SNPs

It has been discovered that a polymorphic variation in a gene encoding ENDO180 is associated with the occurrence of melanoma.


Forty-eight allelic variants located within or nearby the target gene were identified and subsequently allelotyped in melanoma case and control sample sets as described in Examples 1 and 2. One of the SNPs, designated by SNP_ID “AA” in column 1 of Table 16, is novel and was identified through SNP discovery methods using dye terminator sequencing. Methods of diedeoxynucleotide chain termination chemistry derived sequencing are described by Sanger et al. (Proc. Nat. Acad. Sci. USA 74, 5463-5467, 1997). Briefly, terminators are labeled with fluorescent dyes for automated detection. Fluorescein is measured by an argon ion laser in the sequencing instrument, which transfers light energy to a rhodamine acceptor dye. Each acceptor dye emits light at a characteristic wavelength for detection, identifying the nucleotide that terminated extension of the DNA fragment. Novel SNPs are identified when sequence from multiple samples are compared on one or more strands. The forward and reverse primers used to identify AA are GGCTGACTCAACAGCAGAAG and GGAGGCCTGACAGCAAGG, respectively. All of the polymorphic variants from ENDO180 are set forth in Table 16. The chromosome position provided in column four of Table 16 is based on Genome “Build 34” of NCBI's GenBank and dbSNP 119.

TABLE 16relative_positionSnpGenomeDeducedSNP Idin SEQ ID NO: 3ChromChromPositionAllelesLetterIUPACrs24293932241761200674A/CGKrs24293921,8741761202324A/GCYrs24654212,2151761202665T/CTYrs24654107,5851761208035T/CGRrs24603028,0251761208475C/GGSrs24603018,6871761209137A/GTYrs246545912,6031761213053C/GCSrs242939113,8851761214335C/TGRrs201405514,8381761215288C/TGRrs225281415,2691761215719C/AGKAA16,7271761217177G/AGRrs807958218,3881761218838C/TCYrs242939018,7921761219242C/TGRrs720933119,1951761219645A/GGRrs740581519,9251761220375C/TTYrs496870619,9491761220399C/TTYrs496861320,9001761221350C/TTYrs72964721,8471761222297G/ACYrs807298422,4541761222904G/AARrs989644424,1931761224643C/TCYrs496861726,8251761227275A/CAMrs735090728,6671761229117T/GTKrs242938932,2611761232711T/CARrs246030032,2681761232718C/TARrs246545432,8731761233323T/GAMrs246545235,1651761235615C/TGRrs721971135,4491761235899A/CCMrs246544036,8331761237283G/TGKrs242938836,9521761237402T/CARrs378613037,9641761238414T/CTYrs246028939,6541761240104G/AGRrs246542839,7071761240157C/AGKrs74069840,0721761240522G/ACYrs246542541,1641761241614T/CARrs246029041,7671761242217T/CTYrs382653742,7241761243174T/CARrs316975643,1391761243589G/TTKrs155823747,2411761247691T/CARrs242938549,7201761250170T/CGRrs582136850,0361761250486—/TTNrs807253850,8361761251286G/AGRrs225139351,8531761252303C/TGRrs225139151,9461761252396T/CARrs206865757,8641761258314A/GCYrs807251959,4141761259864C/TCYrs722050560,0741761260524C/TCYrs246028865,7211761266171T/CCYrs496873667,9951761268445G/TTK


Assay for Verifying and Allelotyping SNPs


The methods used to verify and allelotype the seventy-six proximal SNPs of Table 16 are the same methods described in Examples 1 and 2 herein. The primers used in these assays are provided in Table 17 and Table 18, respectively.

TABLE 17dbSNP rs #Forward PCR PrimerReverse PCR Primerrs2429393GCCTTCTTGTCATTCTTTTTAGCCGAGATAGTGCCACTCrs2429392CTGGGGGACAGAGTGAAACTCAAGTGTTACTTGAAAGACCrs2465421AACAACTTAGATACCCCCCCCTCAGGAACTAATCATTCTGGrs2465410GACCATATGGTGATAAGGGCGAGAATTTCATAGATGCCAGCrs2460302AAGAGAAAAGGCGTCCTGAGCTCTGCCCGCTTTGTATGTGrs2460301TCCTTTGTCACATGAGGCACAAAGTTGGGTGGATTTGAACrs2465459TCCTGGGTCCAAATCTCAAGTGACTGTCTTTGGAGAATGGrs2429391TCTTTCCACCTATCTCCTCGTCTGTAAAGTGGGTGTGCTGrs2014055TGCAGTCCATGGCTGAAGATTGTGATGCCTTCGTGTCTTGrs2252814AATCTGTCCCTCATGGCTCCACCTATGTGCTCTGCCCTACAAAGAAGGTCTCGCAGTCGTTATAGTGTAGCCTTTTGGTGGGrs8079582TTAAGGGGTGGTGAGTGATGCTACTGAGCTCAAGCAATCCrs2429390GATTCAATTACTCTGGGGCGAGTATGGTCATCCAACCAGCrs7209331TGGGCAAGCAGCTTAGTCTCTCACAACAACCTTGCAAGGGrs7405815AGTGCAGTGGCACAATCTCCGAGGCTGGAGCATGAGAATCrs4968706TCTCATCTCCAGGTTCAAGCGTGGCACACTCATGCCTATAArs4968613TTGCCAAGACCAAGTAAGGGTTCAGGTTCACCTTCTAGGGrs729647TCTGGGAGAGTTTGGAGAGGAACCAACCCACTGGGTTGACrs8072984TAACTAGGATGTCCTTCAGCATCTGGGGCCCGGTGAGATrs9896444AACTGCTTGTGCTCTCATCCTTCTGACATGTCTGTGTCCCrs4968617TCCTCGCTTCACCAAGGCAAGGATGAAGTCATGTACACCCrs7350907TGAACAGGATTGTCCTTAGCCTACTGCCTTTCCCACCATGrs2429389TGTACATGACAGCCAGCCGAATGACTTCTGCCCAGGCTCGrs2460300ACAGACTGATGACTTCTGCCCAGAGTGCGCAGTCAGCTGTrs2465454CACTGATCTAGAACACTCACTCACTGTAGTAGCACCTTCCrs2465452TGATCTCAGGTGATCCACCCTTCAAAAGAACATTACACGGrs7219711TTTTTAAGGTAGAAGAGGTGCAGCCTGGGCGAGAGATTGArs2465440AAGAGGCTGTCCCACAAACCCTCAGGCTGTAATGCCCCTTrs2429388CCTGCCATACCCCTTAATACTATTCAGCTCAGTCCAGCCGrs3786130TAGTAGAGACGGGGTTTCACGAAGCTGCTGCTAATAGATCrs2460289CCCAGACCAAGGCTTGGTAGCTCCCTGTCCATTTGTTAGCrs2465428AGGCACAAGGCCAGGAGAAGACACCTACCAAGCCTTGGTCrs740698GTGGGCTCCCGACACTTTTGAATAGTCAGGATAGGGTGCGrs2465425GGGACAGGGAGTCGGTTCTAAGGAAAGAAGGCACTGAGGGrs2460290AGGCATGAGCCACCACATCAGCCTCAGTTTCCTCGTATGTrs3826537CATCCTGGTGTCAGACATGGAACTGGACAGGCAGCTGGTGrs3169756ACAGAGCTGAGGCTGCCAGCTTCTTTGCAGAGCCCGAGGrs1558237ACCACATGGCTCTCCTGGAATTTGCCTTGTGCCTGCAATGrs2429385CCTGGTTGAAACTACTACTGCGAGATTGCGCCATCGCACTrs5821368CCTACTGGTTGGTTTTCAAGTCACCTTAAAATCTGGTCCCrs8072538TGGCTCATGCCTATAAGCTCCTATGGCCCAGGCTGATTTCrs2251393ACCAAAGCGTTCACAGTTAGATAGATGCTCAAGCCAGACCrs2251391CTTGGGGACGTAGAAAGAAGCATTTCTCAAAGCCAGGTCGrs2068657ACCTTCAGCTAGGCTCTTCGGATTGGATGAAAAGCGAGGCrs8072519GTCTAGCGTCTAGTAGCTTCATTCATGCACTGCCCTGTACrs7220505CCTACTTTCATTTGGGTGCCTGAGCCATGATTGGTGCCACrs2460288ATGCGGCGAGCCAAATAACAGTGTAACCTGTGACCTAAGCrs4968736AGAAAGCAGCAAGGAACGAGTGCTGGTGGCTGATCCTTAG












TABLE 18!Term? ? !dbSNP rs #? Extend Primer? Mix













rs2429393
AGTGTCGCTCTATTGCT
ACT








rs2429392
GAGTGAAACTCGTCTCC
ACT







rs2465421
CAAACAAAAGAAATGTGTAGC
ACT







rs2465410
CAATACCCTCAGTGCCT
ACT







rs2460302
GCTCTGTGTTCTTTGTAC
ACT







rs2460301
ACATGAGGCACAGCTTC
ACT







rs2465459
AGTTCTCTCTCTTCCCC
ACT







rs2429391
ACCCTCTGTAGAAGTCA
ACG







rs2014055
GCTGAAGATGAGGAAGA
ACG







rs2252814
TCCAGTCCCCTGGACAT
CGT







AA
CAGTCGTTACCTGCCACCA
ACG







rs8079582
GGCTCACACCTATAATCC
ACG







rs2429390
CATTTCTAACAAGCCCTC
ACG







rs7209331
CTCATTGCAAATGGAGAAT
ACT







rs7405815
GTTCAAGCGATTCTCATC
ACG







rs4968706
TCAAGCGATTCTCATGC
ACG







rs4968613
CCTCAGCTGCCCTCGGA
ACG







rs729647
GAGGTGACTGGGAGGCCT
ACG







rs8072984
ATTTTGTGGCCCTGTGA
ACG







rs9896444
TGAGGCTTATGGGTTCATCC
ACG







rs4968617
CACCAAGGCAAGTCAAA
ACT







rs7350907
TTGTCCTTAGCTAGCCT
ACT







rs2429389
GCGCAGTCAGCTGTGCA
ACT







rs2460300
CCAGGCTCGCCCAGCAT
ACG







rs2465454
CCAAATTATTTGGACAACC
ACT







rs2465452
CCAAAGTGCTGGGATTA
ACG







rs7219711
GAAGAGGTGTAATTTTTTTTTT
ACT







rs2465440
TCTTAGGAGGTGAGCAG
CGT







rs2429388
CCCTTAATACACTTGCAC
ACT







rs3786130
GCGCCCAGCCCTAGGAC
ACT







rs2460289
GGCTTGGTAGGTGTCAG
ACG







rs2465428
CACACCAGGCAAAGACC
CGT







rs740698
CACTTTTGCCCTGGGCC
ACG







rs2465425
AGTCGGTTCTAGAACAC
ACT







rs2460290
CCACCACATCAGTCTGG
ACT







rs3826537
CATGGAAATGAATGAGCA
ACT







rs3169756
CAGGTGGGAAGAGAGACAC
CGT







rs1558237
TCTCCTGGAACGGAGCC
ACT







rs2429385
TTTTTTTGAGATGGAGTTTC
ACT







rs5821368
CATCACTTTAACTGGAACATA
ACG







rs8072538
GGCCAAGGCAGGTAGAC
ACG







rs2251393
CACAGTTAGATTTTCACCT
ACG







rs2251391
TAGAAAGAAGGGCTGGC
ACT







rs2068657
AGGCTCTTCGCGGACAC
ACT







rs8072519
TCCCTGATCCTCATCTC
ACG







rs7220505
GGTCTCACTCTGTCACA
ACG







rs2460288
GGATGAGGTGAAAACAG
ACT







rs4968736
GCAGTGTGGCCAGGGTC
CGT










Genetic Analysis


Allelotyping results are shown for female (F) and male (M) cases and controls in Table 19 and Table 20, respectively. Allele frequency is noted in the fourth and fifth columns for melanoma pools and control pools, respectively.

TABLE 19FemalesMelanomaChromo-ChromosomeFemale CaseFemale ControlFOddsAssociateddbSNP rs#somePositionAllelesAFAFp-ValueRatioAllelers24293931761200674A/CA = 0.754A = 0.7290.4160.879A  C = 0.245   C = 0.270 rs24293921761202324A/GA = 0.753A = 0.7030.0890.775AG = 0.246G = 0.296rs24654211761202665T/C T = 0.456 T = 0.4770.5351.089C  C = 0.543   C = 0.522 rs24654101761208035T/C T = 0.873 T = 0.8620.6190.905T  C = 0.126   C = 0.137 rs24603021761208475C/G  C = 0.500   C = 0.515 0.6811.059GG = 0.499G = 0.484rs24603011761209137A/GA = 0.482A = 0.5080.4561.106GG = 0.517G = 0.491rs24654591761213053C/G  C = 0.688   C = 0.673 0.6440.934CG = 0.311G = 0.326rs24293911761214335C/T  C = 0.473   C = 0.437 0.3040.867C T = 0.526 T = 0.562rs20140551761215288C/T  C = 0.807   C = 0.785 0.4160.874C T = 0.192 T = 0.214rs22528141761215719C/A  C = 0.562   C = 0.541 0.5620.917CA = 0.437A = 0.458AA1761217177G/AG = 0.893G = 0.9220.2471.416AA = 0.106A = 0.077rs80795821761218838C/T  C = 0.902   C = 0.906 0.8561.044T T = 0.097 T = 0.093rs24293901761219242CAT  C = 0.452   C = 0.415 0.2720.859C T = 0.547 T = 0.584rs72093311761219645A/GA = 0.226A = 0.1540.0110.626AG = 0.773G = 0.845rs74058151761220375C/T  C = 0.078   C = 0.054 0.2140.676C T = 0.921 T = 0.945rs49687061761220399C/T  C = 0.165   C = 0.063 ˜0.0010.340C T = 0.834 T = 0.936rs49686131761221350C/T  C = 0.132   C = 0.100 0.1570.731C T = 0.867 T = 0.899rs7296471761222297A/GA = 0.700A = 0.7110.0351.053GG = 0.299G = 0.288rs80729841761222904G/AG = 0.216G = 0.1370.0030.580GA = 0.783A = 0.862rs98964441761224643C/T  C = 0.893   C = 0.932 0.0591.649T T = 0.106 T = 0.067rs49686171761227275A/CA = 0.695A = 0.7320.2301.201C  C = 0.304   C = 0.267 rs73509071761229117T/G T = 0.774 T = 0.8450.0141.586GG = 0.225G = 0.154rs24293891761232711T/C T = 0.011 T = 0.0260.2272.403C  C = 0.989   C = 0.974 rs24603001761232718C/T  C = 0.983   C = 0.973 0.3850.609T T = 0.167 T = 0.027rs24654541761233323T/G T = 0.186 T = 0.0280.4711.533TG = 0.981G = 0.972rs24654521761235615C/T  C = 0.047   C = 0.039 0.6710.826C T = 0.952 T = 0.960rs72197111761235899A/CA = 0.809A = 0.7900.5120.891A  C = 0.190   C = 0.209 rs24654401761237283G/TG = 0.672G = 0.6530.5680.918G T = 0.327 T = 0.346rs24293881761237402T/C T = 0.030 T = 0.0370.6601.227C  C = 0.969   C = 0.962 rs37861301761238414T/C T = 0.733 T = 0.7490.6601.090C  C = 0.266   C = 0.250 rs24602891761240104G/AG = 0.045G = 0.0300.3680.655GA = 0.954A = 0.969rs24654281761240157C/A  C = 0.016   C = 0.021 0.6851.353AA = 0.984A = 0.978rs7406981761240522G/AG = 0.142G = 0.1130.2910.766GA = 0.857A = 0.886rs24654251761241614T/C T = 0.027 T = 0.0320.7251.210C  C = 0.972   C = 0.967 rs24602901761242217T/C T = 0.039 T = 0.0280.5530.716T  C = 0.960   C = 0.971 rs38265371761243174T/C T = 0.644 T = 0.6760.3791.154C  C = 0.355   C = 0.323 rs31697561761243589G/TG = 0.067G = 0.0420.1250.609G T = 0.932 T = 0.957rs15582371761247691T/C T = 0.070 T = 0.0630.7260.892T  C = 0.929   C = 0.936 rs24293851761250170T/C T = 0.956 T = 0.9520.8110.913T  C = 0.043   C = 0.047 rs58213681761250486—/T— = 0.375  — = 0.396  0.5241.091T T = 0.624 T = 0.603rs80725381761251286G/AG = 0.016G = 0.0220.6191.379AA = 0.984A = 0.978rs22513931761252303C/T  C = 0.222   C = 0.203 0.5320.896C T = 0.777 T = 0.796rs22513911761252396T/C T = 0.146 T = 0.0990.0780.648T  C = 0.853   C = 0.900 rs20686571761258314A/GA = 0.061A = 0.0620.9601.016GG = 0.938G = 0.937rs80725191761259864C/T  C = 0.763   C = 0.828 0.0321.496T T = 0.236 T = 0.171rs72205051761260524C/T  C = 0.714   C = 0.701 0.7040.942C T = 0.285 T = 0.298rs24602881761266171T/C T = 0.168 T = 0.1980.2501.221C  C = 0.831   C = 0.801 rs49687361761268445G/TG = 0.302G = 0.3060.8841.021T T = 0.697 T = 0.693









TABLE 20










Males























Melanoma



Chromo-
Chromosome

Male Case
Male Control
M
Odds
Associated


dbSNP rs#
some
Position
Alleles
AF
AF
p-Value
Ratio
Allele





rs2429393
17
61200674
A/C
A = 0.705
A = 0.726
0.524
1.108
C







  C = 0.294 


  C = 0.273 



rs2429392
17
61202324
A/G
A = 0.692
A = 0.720
0.412
1.142
G






G = 0.307
G = 0.279


rs2465421
17
61202665
T/C
 T = 0.490
 T = 0.466
0.496
0.907
T







  C = 0.509 


  C = 0.533 



rs2465410
17
61208035
T/C
 T = 0.848
 T = 0.875
0.263
1.259
C







  C = 0.151 


  C = 0.124 



rs2460302
17
61208475
C/G

  C = 0.545 


  C = 0.527 

0.618
0.933
C






G = 0.454
G = 0.472


rs2460301
17
61209137
A/G
A = 0.536
A = 0.489
0.178
0.830
A






G = 0.463
G = 0.510


rs2465459
17
61213053
C/G

  C = 0.659 


  C = 0.655 

0.905
0.982
C






G = 0.340
G = 0.344


rs2429391
17
61214335
C/T

  C = 0.503 


  C = 0.437 

0.063
0.768
C






 T = 0.496
 T = 0.562


rs2014055
17
61215288
C/T

  C = 0.812 


  C = 0.812 

0.994
0.999
C






 T = 0.187
 T = 0.187


rs2252814
17
61215719
C/A

  C = 0.605 


  C = 0.539 

0.064
0.762
C






A = 0.394
A = 0.460


AA
17
61217177
G/A
G = 0.908
G = 0.943
0.067
1.685
A






A = 0.091
A = 0.056


rs8079582
17
61218838
C/T

  C = 0.908 


  C = 0.940 

0.095
1.594
T






 T = 0.091
 T = 0.059


rs2429390
17
61219242
C/T

  C = 0.477 


  C = 0.390 


0.011

0.701
C






 T = 0.522
 T = 0.609


rs7209331
17
61219645
A/G
A = 0.167
A = 0.100

0.009

0.558
A






G = 0.832
G = 0.899


rs7405815
17
61220375
C/T

  C = 0.067 


  C = 0.045 

0.212
0.658
C






 T = 0.932
 T = 0.954


rs4968706
17
61220399
C/T

  C = 0.096 


  C = 0.071 

0.293
0.725
C






 T = 0.903
 T = 0.928


rs4968613
17
61221350
C/T

  C = 0.111 


  C = 0.081 

0.155
0.706
C






 T = 0.888
 T = 0.918


rs729647
17
61222297
A/G
A = 0.660
A = 0.740

0.033

1.472
G






G = 0.339
G = 0.259


rs8072984
17
61222904
G/A
G = 0.161
G = 0.098

0.013

0.567
G






A = 0.838
A = 0.901


rs9896444
17
61224643
C/T

  C = 0.917 


  C = 0.965 


0.024

2.511
T






 T = 0.082
 T = 0.034


rs4968617
17
61227275
A/C
A = 0.681
A = 0.757

0.018

1.463
C







  C = 0.318 


  C = 0.242 



rs7350907
17
61229117
T/G
 T = 0.804
 T = 0.845
0.165
1.326
G






G = 0.195
G = 0.154


rs2429389
17
61232711
T/C
 T = 0.055
 T = 0.031
0.142
0.548
T







  C = 0.944 


  C = 0.968 



rs2460300
17
61232718
CAT

  C = 0.961 


  C = 0.974 

0.339
1.571
T






 T = 0.038
 T = 0.025


rs2465454
17
61233323
T/G
 T = 0.056
 T = 0.027
0.088
0.479
T






G = 0.943
G = 0.972


rs2465452
17
61235615
C/T

  C = 0.057 


  C = 0.032 

0.117
0.545
C






 T = 0.942
 T = 0.967


rs7219711
17
61235899
A/C
A = 0.789
A = 0.793
0.903
1.023
C







  C = 0.210 


  C = 0.206 



rs2465440
17
61237283
G/T
G = 0.672
G = 0.625
0.173
0.813
G






 T = 0.327
 T = 0.374


rs2429388
17
61237402
T/C
 T = 0.053
 T = 0.026
0.096
0.492
T







  C = 0.946 


  C = 0.973 



rs3786130
17
61238414
T/C
 T = 0.754
 T = 0.796
0.272
1.270
C







  C = 0.245 


  C = 0.203 



rs2460289
17
61240104
G/A
G = 0.039
G = 0.044
0.806
1.123
A






A = 0.960
A = 0.955


rs2465428
17
61240157
C/A

  C = 0.047 


  C = 0.028 

0.236
0.590
C






A = 0.952
A = 0.971


rs740698
17
61240522
G/A
G = 0.130
G = 0.083
0.081
0.602
G






A = 0.869
A = 0.916


rs2465425
17
61241614
T/C
 T = 0.040
 T = 0.019
0.237
0.470
T







  C = 0.960 


  C = 0.981 



rs2460290
17
61242217
T/C
 T = 0.084
 T = 0.048
0.105
0.551
T







  C = 0.915 


  C = 0.951 



rs3826537
17
61243174
T/C
T =   
T =   






C =    
C =    


rs3169756
17
61243589
G/T
G = 0.050
G = 0.031
0.250
0.619
G






 T = 0.949
 T = 0.968


rs1558237
17
61247691
T/C
 T = 0.071
 T = 0.068
0.891
0.960
T







  C = 0.928 


  C = 0.931 



rs2429385
17
61250170
T/C
 T = 0.934
 T = 0.950
0.366
1.342
C







  C = 0.065 


  C = 0.049 



rs5821368
17
61250486
—/T
— = 0.399  
— = 0.407  
0.808
1.034
T






 T = 0.600
 T = 0.592


rs8072538
17
61251286
G/A
G = 0.029
G = 0.023
0.646
0.786
G






A = 0.970
A = 0.976


rs2251393
17
61252303
C/T

  C = 0.215 


  C = 0.178 

0.187
0.793
C






 T = 0.784
 T = 0.821


rs2251391
17
61252396
T/C
T =   
T =   






C =    
C =    


rs2068657
17
61258314
A/G
A = 0.072
A = 0.060
0.557
0.832
A






G = 0.927
G = 0.939


rs8072519
17
61259864
C/T

  C = 0.793 


  C = 0.848 


0.042

1.457
T






 T = 0.206
 T = 0.151


rs7220505
17
61260524
C/T

  C = 0.710 


  C = 0.707 

0.944
0.989
C






 T = 0.289
 T = 0.292


rs2460288
17
61266171
T/C
 T = 0.169
 T = 0.187
0.505
1.131
C







  C = 0.830 


  C = 0.812 



rs4968736
17
61268445
G/T
G = 0.277
G = 0.283
0.850
1.029
T






 T = 0.722
 T = 0.716









Allelotyping results were considered particularly significant with a calculated p-value of less than or equal to 0.05 for allelotype results. These values are indicated in bold. The assay failed for those SNPs in which the allele frequency is blank. The combined allelotyping p-values for males and females were plotted in FIG. 2A and separately for females and males in FIGS. 2B and 2C, respectively. The position of each SNP on the chromosome is presented on the x-axis. The y-axis gives the negative logarithm (base 10) of the p-value comparing the estimated allele frequency in the case group to that of the control group. The minor allele frequency of the control group for each SNP designated by an X or other symbol on the graphs in FIGS. 2A, 2B and 2C can be determined by consulting Table 19 or 20. By proceeding down the Table from top to bottom and across the graphs from left to right the allele frequency associated with each symbol shown can be determined.


To aid the interpretation, multiple lines have been added to the graph. The broken horizontal lines are drawn at two common significance levels, 0.05 and 0.01. The vertical broken lines are drawn every 20 kb to assist in the interpretation of distances between SNPs. Two other lines are drawn to expose linear trends in the association of SNPs to the disease. The light gray line (or generally bottom-most curve) is a nonlinear smoother through the data points on the graph using a local polynomial regression method (W. S. Cleveland, E. Grosse and W. M. Shyu (1992) Local regression models. Chapter 8 of Statistical Models in S eds J. M. Chambers and T. J. Hastie, Wadsworth & Brooks/Cole.). The black line (or generally top-most curve, e.g., see peak in left-most graph just to the left of position 92150000) provides a local test for excess statistical significance to identify regions of association. This was created by use of a 10 kb sliding window with 1 kb step sizes. Within each window, a chi-square goodness of fit test was applied to compare the proportion of SNPs that were significant at a test wise level of 0.01, to the proportion that would be expected by chance alone (0.05 for the methods used here). Resulting p-values that were less than 10−8 were truncated at that value.


Finally, the exons and introns of the genes in the covered region are plotted below each graph at the appropriate chromosomal positions. The gene boundary is indicated by the broken horizontal line. The exon positions are shown as thick, unbroken bars. An arrow is placed at the 3′ end of each gene to show the direction of transcription.


Example 7
CDK10 Proximal SNPs

It has been discovered that a polymorphic variation (rs8404) in the untranslated region of a gene encoding cyclin-dependent kinase 10 (CDK10) is associated with the occurrence of melanoma. See Table 3. Subsequently, one-hundred-three allelic variants located within or nearby the target gene were identified and subsequently allelotyped in melanoma case and control sample sets as described in Examples 1 and 2. Several SNPs, designated by SNP_ID “AA”, “AB”, and “AC” in column 1 of Table 10, are novel and were identified through SNP discovery methods using dye terminator sequencing. Methods of diedeoxynucleotide chain termination chemistry derived sequencing are described by Sanger et al. (Proc. Nat. Acad. Sci. USA 74, 5463-5467, 1997). Briefly, terminators are labeled with fluorescent dyes for automated detection. Fluorescein is measured by an argon ion laser in the sequencing instrument, which transfers light energy to a rhodamine acceptor dye. Each acceptor dye emits light at a characteristic wavelength for detection, identifying the nucleotide that terminated extension of the DNA fragment. Novel SNPs are identified when sequence from multiple samples are compared on one or more strands. The forward and reverse primers used to identify AA are TTCTTCTTGCCCAGATGGAG and GGCTAGCTTTGGGAGATGTTT, respectively. The forward and reverse primers used to identify AB are CACAGAACTGAGTGGCTGGA and AGAGCTGAGCCTGCAGTGA, respectively. The forward and reverse primers used to identify AC are GGAGGCTGTGTCCTGTGGAG and CTATGGGAGGCAGGAGCTGA, respectively. The polymorphic variants are set forth in Table 21. The chromosome position provided in column four of Table 21 is based on Genome “Build 34” of NCBI's GenBank and dbSNP build “119”.

TABLE 21PositionAlleledbSNPChromo-in SEQChromosomeAllelePresent inIUPACrs#someID NO.: 4PositionVariantsSEQ ID NO: 4Coders4608791623489456684T/CCYrs4609841651989456969T/CCYrs2437957162,99389459443C/TGRrs3815949163,26189459711C/GCSrs2437956163,59689460046G/ATYrs2437955163,62089460070G/TCMrs2434872164,26089460710C/GCSrs467357164,74289461192G/AARrs258337168,05589464505A/TTWrs258336168,17689464626T/GTKrs258335168,28989464739A/GGRrs164752169,73589466185G/AGRrs1546631613,38089469830T/CTYrs1647531614,94089471390T/CCYrs2583321615,02889471478G/AGRrs20106231616,37089472820C/TARrs1872831616,68189473131G/AARrs2583301616,91989473369T/GTKrs2583281617,65989474109C/TCYrs4599201618,17289474622T/CTYrs1662971618,53089474980G/AARrs2583181619,39589475845T/CTYrs2583171619,58389476033C/TCYrs1718051620,95989477409G/AARrs20819841621,27189477721C/TCYrs4645861621,43389477883C/TARrs4603191621,43889477888G/TGKrs4477351621,69489478144A/GTYrs23770581622,17689478626T/CARrs6048681623,52289479972G/ATYrs21181931627,24889483698C/TGRrs4670351627,63089484080C/GCSrs21154011627,95489484404C/TTYrs4173231633,62289490072T/GTKrs23772331634,24789490697C/GGSAA1636,58989493039G/CCSrs97461141638,67289495122C/GGSrs37517001639,53989495989G/AGRrs22779051639,84689496296A/GARrs3978911640,37689496826C/GGSrs37946381640,78189497231G/TCMrs37946371640,91589497365G/ACYrs2583241641,60089498050C/ATKrs2583231642,03989498489C/GGSrs20758801642,56189499011A/GARrs2583221643,24889499698C/TARrs2583211643,81889500268C/TARrs1647441644,11389500563A/CTKrs38338291644,12689500576A/—CNAB1644,54489500994T/CCYrs1647431644,77689501226A/GTYrs1647421645,07089501520C/TARrs1872821646,31889502768T/CARrs4655071647,83789504287C/GCSrs21629431647,99789504447G/ACYAC1648,30489504754G/CCSrs19464821649,75589506205T/CTYrs84041649,99289506442C/TTYrs10458141650,26089506710G/ACYrs42473531650,55989507009T/CTYrs4627691650,60889507058A/GTYrs38036901650,70489507154C/GGSrs37516961650,96489507414A/TAWrs99728641652,54289508992A/GGRrs105672261653,30789509757TTT/—TNrs4174141653,49389509943A/GCYrs1546611653,91689510366G/CGSrs47857041659,71589516165G/AARrs47855841659,72989516179C/TTYrs47855851659,75089516200C/TCYrs47857051659,75989516209T/CCYrs47855861659,76189516211G/AGRrs47855871659,96489516414A/GGRrs20169681660,25189516701C/GCSrs37516931661,74189518191C/TGRrs38096461674,20089530650A/GCYrs47855901677,51289533962G/CGSrs20991051680,90089537350A/GTYrs47857081683,91489540364C/TCYrs47857101684,10189540551T/CCYrs47855911684,85889541308C/TCYrs47855921684,86089541310C/GGSrs47855931684,97289541422C/TCYrs47857111685,29189541741C/GCSrs47857121685,29989541749G/CCSrs47857131685,44389541893G/CCSrs47857141685,74989542199C/ACMrs47855941685,86189542311C/GGSrs38036891687,40389543853A/GCYrs47857151687,48089543930G/AARrs37642581688,35989544809C/GCSrs47857161688,47289544922T/CTYrs47857171688,47889544928G/AARrs20770011689,43589545885C/TGRrs23770491690,31389546763A/CTKrs20035221690,63189547081C/GCSrs12301692,20089548650C/TCYrs18003591692,60689549056A/GARrs10616461693,32289549772T/CGRrs18003581696,66489553114C/TTYrs22863921696,89489553344C/TARrs20749041698,89189555341T/CARrs20749031699,00889555458A/GTY


Assay for Verifying and Allelotyping SNPs


The methods used to verify and allelotype the one-hundred-three proximal SNPs of Table 21 are the same methods described in Examples 1 and 2 herein. The primers and probes used in these assays are provided in Table 22 and Table 23, respectively.

TABLE 22dbSNP rs #Forward PCR primerReverse PCR primerrs460879TCCAGCCTCTAACCCACAACCGAAACGCTGTGTCACCAACrs460984ATGCTCTGGACAACAGGTGGTGAGGGAACGAAAAGTGCAGrs2437957ATCCAGCTTCCCTCAACCTCCAGCTCAGAGTTGACGGAAGrs3815949TTTTTCTGCAGAAGGGCCTGCACACACCACCAAGCTCTTCrs2437956TCAACACTGGGCTCCTAGGGTCTCCTGACCTCGTGATCTGrs2437955TCAACACTGGGCTCCTAGGGTCTCCTGACCTCGTGATCTGrs2434872TAGACACCAGCTCTGTAGGGATTACAGGCATGAGCCACTGrs467357TCATCTCTGTGCTCTGGACGTGTGTGGTGCAGCCCAGAATrs258337TAGAATCCAAAAGATGGTGGTGCTGGGATTACAGGTGAACrs258336CTTTGAGGAGAAAGAGTCTGTTAAAGCCAGGGTTTGAGGCrs258335TCCTGACCTCAAGTGATCTGAAAGAGCTGATTTCCAGCGGrs164752AAGTGCTGGAATTACAGCCGAAAATATGCTGGCCAGGCCGrs154663ATTGCTCGAACCTTGAACCGGACAGAGTCTCACTCTGTAGrs164753AGTGAGAAGAACAGAAGGGCGCCAAGCCACATCAATATCCrs258332TGTGTCCTCACCCAAATCTCCCTGCCCCCATAATTCAATCrs2010623CCATGACGCCCAGCTAATTTAACCAGGAGTTTGAGGCCAGrs187283CCGGGTTCAAGCAATTCTTCAATTACCCAGGCATGGTGGCrs258330GCTTTTTTCACTCAGCATGGGGTAGAGTATACTTAATTGCrs258328TCAGGTTTCACCATGTTGCCCAGAACTTTGGGAAGCCTAGrs459920AACAGCCATGTGTAACCCTCTAATCGCAGGAGAGACACACrs166297ACTTCATTCCTTGAACCGGGACTGCATTCTCATGAGGAGCrs258318TGCGTCTGGCCTAATATTTCCACTATTGAAATTTTCTAGTCrs258317TGAACTCCTGACCTCAAGTGGCCAAGTGTAGTGATTCACGrs171805ACCATGGTCTGCTTGGCAACGCCTTTTGTTATGCCCAGACrs2081984TTCCAAGTAGCTGGGACCACTATTACACTGGAACAGCTCGrs464586GACCTTCAAGATCCTGTCTCATAGCCTCATCTGTGTCAGCrs460319GACCTTCAAGATCCTGTCTCATAGCCTCATCTGTGTCAGCrs447735ATGGCTTGGCTTGTCTGTACCACAGCAGCTTCTGCTATAGrs2377058GGGTTAGAAGACTCAGTCACTACCAAGTGTCCCACAAAGCrs604868CAACCCTGGTGTACCCTCCCTGCCCCCGCCGTGAGAAArs2118193TTTTTGAGACGGAGTCTCGCAGGAGAATGGCTGAACCTGGrs467035CGTGTCTGTTTCCACTCTTGCCCTGTGTAGAAAAGAAGGCrs211540TTGGCATCTTTGGGCGTTTTGCAACCCGCTTCAGCCTTGAGrs417323GAGGACTGAAGAAAGAATTTGGCGAGACTCTGTCTCAAATArs2377233TCTCTTTTTGGACTTCAGGGTCCCACAGTGCTAGGATTACAAGGTCCTGCAGATAACAATGCAGCCGTAGGTATCAGACATGrs9746114TGAATGGAGTCTAGGTCTGGTGTTTATGGCCAGTTTTGCGrs3751700TCAGGTCTTCCATGAGTGAGTTCAAGCTTAGCTTCTGGGCrs2277905TCCTCCAGTTGGGAGTCCTTCACCTGGAAGACTCTCTCACrs397891CGGGAGCGTCTCTTGGTAACCAGGCGCGAAAGCTCCTTCrs3794638AGCCTGCTCTTCCCAAGTCCCGGTCCCTGGAGATCTGAGrs3794637GAGCCTTCAAGCTCGAACTCAGCCGCGAGCCACTTGTTTGrs258324AAGCAGAGGATGTGAGAAGGAAGCATCTGGCCTGTCACTCrs258323CACGCCCAGCTAATTTTTTCAGTCAGGAGATCTAGAGCACrs2075880AAGTGCTGGGATTACAAGCGAAAGCATTGGCCCAGTATCGrs258322TCTGAAACAGGGCAGAAACCGGATGCTTACGTTTACCCAGrs25832TTTGGCAACAGAGTGAGACCCAGCTTCCAGAATGACCTACGrs164744TCAGTCCCCACTCTGTGCAAACAGAGCAAAGTTGGGCACCrs3833829ATGCCCAGTGACACTCAGTCCCTGCCCAGTTCAAATGAGGABATGGAGACTGCAGAGCCCTCTGGTGTGGCCCAGCACATCArs164743AAAAGTGCTTATTGGGCCGGAACTGCCGACCTCAGGTGATrs164742AACGAGCGAAACTCCGTCTCTGATAGCAGGTGCTGCAGTGrs187282TGGGAACCCCTGAGCTTTTAGTTGGAAGCACAAATTCGGCrs465507AAGCACCCACCTCAATACGGTCCGAGTTGGGACAGGTTTCrs2162943TGAGCTCAGGAACCAGGTGACCAGCATCGACATGTGGTGAGACTGACTGCAGCCAGACCTTTGTGCCTGTCCTTCACAGTGTCrs1946482TCCTCGCTATGTTGGAAATGACCACAACTCAAAGACAGCGrs8404AAGGTATGGGGTGGGAGCGCCTCCTGTTGGGTCCTCrs1045814AGAGGGGCTGTCACAGTGGATGGGTCCAGTGGTAATGGTGrs4247353AGCTGCTGTGACACCCAAGGAAGCTCTGCTGGAGCCAATCrs462769AAGCCAGGTCAGCCGTGCATCCTCAAGGGCCTGTTGGTGrs3803690ACACTCGTGTGGAGAATCCCTCCCTGCAGTACGGAGTACGrs3751696ATACAGAGAGTCCCACCTCCTCTGGTTGCCTTGTCCCTGGrs9972864TACAAGCGTGAGCCACTGTGACTGTACTCCAGACAGAGCGrs10567226TCCATCCTGGTGACAGAGTGACAGCCGTGGGACGGTAGAArs417414CCACACCCAGCTCATTTTTGGAGATTGACACCAGCCTAACrs154661CCCTTGAGGCCTCCCAGGGGGCCAGGAAGCAGCAGAACrs4785704CACGCCCGGCTGATTTTTTGCACGAGGTCAGGAGATCAAGrs4785584TAATCCCAGCACTTTGGGAGGGTTTCACCGTGTTAGCCAGrs4785585CCTGTAATCCCAGCACTTTGAGGATGGTCTTGATCTCCTGrs4785705AGGATGGTCTTGATCTCCTGCCTGTAATCCCAGCACTTTGrs4785586ATCTCCTGACCTCGTGATCCAAATCCTACAAATGGCCGGGrs4785587AACAAATCCTGGCGTGGGAGGGATACAGCTGAGCCAGGACrs2016968GACACTTCTGTCTCCCTATGAAAAGCAGTAGCTGTGGACCrs3751693ATCAGGTACCTGCATGGATGGCAGACACTACCTTCTGGAGrs3809646AAAAATGCACGAACGCACGGTAGAGGAGATGTAGCAGGAGrs4785590ACCTCTTGCTGGTGCTAGACCTAGAGCAGTGGAGACATTCrs2099105ATCAGAGGCTGCATAGCACCGATGACTAGAAGGGAGACTGrs4785708TGTTCATGCTGAAGCTGCTGGAAAAGAGAAACGGGAGCAGrs4785710TTGTTCCCAGGAGTCATGGCAGGTGTCCTGGGATTCAGACrs4785591ATTGGGTGGCCCCTTGTTTGGAGGAGGTGAGTGGTCATTGrs4785592ATTGGGTGGCCCCTTGTTTGGAGGAGGTGAGTGGTCATTGrs4785593CGCACTGAGAAGAACTGTTGGGCTAGGTCATGAAAACCAGrs4785711TAAAGAGTGCATTGAGGCCGAACTCCTGACCTCATGATCCrs4785712CAAAGTGCTGGGATTACAGGAACAACATGAGACCCTGTCGrs4785713AAGTGAGCCTCTCACCTCAGTAGTTTACACTGCCGAGTCCrs4785714CTGTGAAGGACTGAAAGCTCTGAGAGCAGTGGTGAGAGAArs4785594GCACTTGTGGTTCCTTTCTGTATCGTGGTGAAGAAGTTCCrs3803689GTGTTTCTGCAGCCTAACTCTAGGACTGCTTCAGGTCTCGrs4785715TGAAGCAGTCCTAGCCAGTGAAGAAACTTCTAGACCCCCCrs3764258TTCATGCCTTCTGCCTTGCCATCTCAGCCTTCCAAAGTGCrs4785716ATGAAGGCACACCAGCTCTGTAATGACACGACCTACACACrs4785717ATGAAGGCACACCAGCTCTGCGGTCTTAATGACACGACCTrs2077001AGGCTAAGGCAGTAGGATTGTTTGGTAAAGACGGCCTCACrs2377049CACAAAGTGCTCAATCCCAGACCATATTGATTGGGCTGGGrs2003522CTGAGATTGGGCTGTTGCACCTTGGCATTTTACTCTACGCrs1230TCTCCTCGACTGCTTTAGTGCCAGTGGTTTATTTTCCCGCrs1800359AAAGAGCTTCTCACACGTGGGGCAGCTGTCAATTCTCATGrs1061646ATAGGCAGAGATGTCCAGAGAAAAAGCGAAAGGCAGCAGCrs1800358TTCTCTCTGTCCCAGTTTCCAATCGCAAAGTGCAGTGCAGrs2286392CAGGTCCACGTGAGAGTGTGCAGTCAGCAGCTCTCAGAACrs2074904TGTAGTGGCCTGTAGGAGCAAGATGAGGGTGGCTGAGATGrs2074903CGTCAATTAAGGCTCAAAGCCCTTCTCCAAATTCCACGTC












TABLE 23













Term












dbSNP rs #
Extend Primer
Mix
















rs460879
ACCAACCCCACGCTCTG
ACT








rs460984
CTTGTGAAGCTGAGTGG
ACT







rs2437957
CGGAAGGTGGATCAGCG
ACG







rs3815949
TCCAGCTGCTTTCCCCC
ACT







rs2437956
TCGGCCTCCCAAAGAGC
ACG







rs2437955
ACAGGCGTGAGCCACCG
CGT







rs2434872
GCGGATCCTATCATTTTTA
ACT







rs467357
GTGCAGCCCAGAATGGTTTC
ACG







rs258337
AAAGAATCCCAAGGGAC
CGT







rs258336
CTGGGCCAATTTAAAAGT
ACT







rs258335
TTTCCAGCGGTACAGTC
ACT







rs164752
GCTCAGGCCTGTAACCC
ACG







rs154663
GTGCGATCTCGCTCACC
ACT







rs164753
TCCTAACAGCTGCCACA
ACT







rs258332
CCATAATTCAATCACCTCC
ACG







rs2010623
GCCAACGTGGCGAAACC
ACG







rs187283
GGCGCACACCTGTAGTC
ACG







rs258330
AAGAGAAATGGAACACTTC
ACT







rs258328
CAGGCAGATCACTTGAG
ACG







rs459920
CCTCCCTCTTCCTCATTCC
ACT







rs166297
CTCTTCTAAGCCCATTC
ACG







rs258318
GACTTATTTCATCTTCCTCA
ACT







rs258317
ATTCACGCCTGTAATCC
ACG







rs171805
AGGGCTCCTTGGTCTAG
ACG







rs2081984
GGCACCTGGGTGACTTG
ACG







rs464586
CTAAGCACTGAGCGATA
ACG







rs460319
CTGCACTAAGCACTGAG
CGT







rs447735
TTCTGCTATAGGTCTCTGACA
ACT







rs2377058
CCACAAAGCTGTAAAACA
ACT







rs604868
CGCCGTGAGAAACTGCAG
ACG







rs2118193
GCAGTGAGCTGGGATCG
ACG







rs467035
AAGAAGGCACCAGACTC
ACT







rs2115401
CCTTGAGCACTGGAGTC
ACG







rs417323
ACACTTTTTATATTTGACAAACTT
ACT







rs2377233
GCTAGGATTACAGGTGT
ACT







AA
TTGGCCTTTTGCAGTACCTC
ACT







rs9746114
GGCTGTGGGCTGTTGGGAA
ACT







rs3751700
CAGCTATGTCAGCATTC
ACG







rs2277905
CTCTCACCTGGAGGACC
ACT







rs397891
AAGCTCCTTCCCGGGCTTC
ACT







rs3794638
GGCTCAGGGATGCCTCG
CGT







rs3794637
TGTGGGGAGAATTTACA
ACG







rs258324
TGATGGCCTAGTCTCAA
CGT







rs258323
TAGAGCACCCTGACTAA
ACT







rs2075880
CCACCTTTTCTCATCAGA
ACT







rs258322
ACCCAGAAATGGTACAA
ACG







rs258321
GTGGTGTTTTTGGTTTTTT
ACG







rs164744
CCCCTGCCCAGTTCAAA
ACT







rs3833829
CCGAGACGCGTTCCCCC
ACT







AB
CCCTCCTGGCAGACAGCT
ACT







rs164743
GTGCTGGGATTACAGGC
ACT







rs164742
CCCCACCTCAGGGAGAA
ACG







rs187282
CACAAATTCGGCTGAGGCCT
ACT







rs465507
GGACAGGTTTCCAGGCCA
ACT







rs2162943
CATGTGGTGAGGAGATA
ACG







AC
CCTGCCAGTGTGAGGCAC
ACT







rs1946482
CACCCACCACTCCTCCC
ACT







rs8404
CGAGACTACCAGGAGAGCCC
ACG







rs1045814
CTGTCACAGTGGATGCACC
ACG







rs4247353
CTGACCTGGCTTCGGGG
ACT







rs462769
ACCCCAAGCTCTGCTGG
ACT







rs3803690
GAGTACGGGGCTCAGGA
ACT







rs3751696
GGTTGGCCTGGACTCTCCA
CGT







rs9972864
GCCACTGTGCCCGGCCTA
ACT







rs10567226
GACTCCGTCTCAAAAAAAAAAAAA
ACT







rs417414
CAACATGGTGAAACCCC
ACT







rs154661
GTCCAAACCAGGGCTGTCC
ACT







rs4785704
ATCAAGACCATCCTGGC
ACG







rs4785584
GTTAGCCAGGATGGTCT
ACG







rs4785585
CTCCTGACCTCGTGATC
ACG







rs4785705
CAGCACTTTGGGAGGCC
ACT







rs4785586
CCCAGCACTTTGGGAGG
ACG







rs4785587
TGAGCCAGGACGTTCCC
ACT







rs2016968
CTCTGTGCTTGCACCCT
ACT







rs3751693
GTGTCCTTCTGTTCGTT
ACG







rs3809646
CCAAAATCCGAGTGACG
ACT







rs4785590
TGCTGAAAACCCTGCCC
ACT







rs2099105
CTCAAGTTCTCCCCACC
ACT







rs4785708
GAGCAGTGGAGGAGGCC
ACG







rs4785710
GGATTCAGACTCAACCC
ACT







rs4785591
AGTGGTCATTGTGGGAA
ACG







rs4785592
TGGTCATTGTGGGAACG
ACT







rs4785593
GGGACAGGGAGAGAAGG
ACG







rs4785711
GGGATTACAGGCGTGAG
ACT







rs4785712
AGAGTGCATTGAGGCCG
ACT







rs4785713
TCATGTTGGCAGTCCCA
ACT







rs4785714
GGTGAGAGAATCCGTAT
CGT







rs4785594
GAAGTTCCTTTCTGTTCTT
ACT







rs3803689
TCAAAGTGTCCTTGGGC
ACT







rs4785715
CAAGTGAAAGTCTGGCC
ACG







rs3764258
GATTACAGGTGTGAGCA
ACT







rs4785716
GACCTACACACATGTGAA
ACT







rs4785717
GACACGACCTACACACA
ACG







rs2077001
ACGGCCTCACTATGTTG
ACG







rs2377049
CTGGTCTTGAACTGACT
ACT







rs2003522
TCTTGCTCTGTTGCCCA
ACT







rs1230
CGCAAACGCTGAGTGACT
ACG







rs1800359
CAATTCTCATGTCCCCCAC
ACT







rs1061646
GGTCTGCAACACCAAGAA
ACT







rs1800358
CCAGTCCGGGTTGGGTGC
ACG







rs2286392
TCAGAACGCCAGGATGC
ACG







rs2074904
CCTCCGCTGCCCCAGCC
ACT







rs2074903
CCATGAGTGTGGGTAATAAACT
ACT











Genetic Analysis


Allelotyping results are shown for female (F) and male (M) cases and controls in Table 24 and Table 25, respectively. Allele frequency is noted in the fourth and fifth columns for melanoma pools and control pools, respectively.

TABLE 24FemalesMelanomaChromo-ChromosomeFemaleFemaleFOddsAssociateddbSNP rs#somePositionAllelesCase AFControl AFp-ValueRatioAllelers4608791689456684T/C T = 0.492 T = 0.5950.0371.520C  C = 0.508   C = 0.405 rs4609841689456969T/C T = 0.780 T = 0.8460.0431.552C  C = 0.219   C = 0.153 rs24379571689459443C/T  C = 0.501   C = 0.508 0.8491.026T T = 0.498 T = 0.491rs38159491689459711C/G  C = 0.002   C = 0.043 0.00118.006GG = 0.998G = 0.957rs24379561689460046G/AG = 0.754G = 0.7560.9621.009AA = 0.245A = 0.243rs24379551689460070G/TG = 0.846G = 0.7530.0160.554G T = 0.153 T = 0.246rs24348721689460710C/G  C = 0.530   C = 0.546 0.6201.069GG = 0.469G = 0.453rs4673571689461192G/AG = 0.649G = 0.6590.7791.044AA = 0.350A = 0.340rs2583371689464505A/TA = 0.571A = 0.6430.0471.351T T = 0.428 T = 0.356rs2583361689464626T/G T = 0.618 T = 0.5550.0930.771TG = 0.381G = 0.444rs2583351689464739A/GA = 0.769A = 0.7520.6020.911AG = 0.230G = 0.247rs1647521689466185G/AG = 0.128G = 0.0570.0030.411GA = 0.871A = 0.942rs1546631689469830T/C T = 0.196 T = 0.2080.6791.082C  C = 0.803   C = 0.791 rs1647531689471390T/C T = 0.864 T = 0.8320.2190.780T  C = 0.135   C = 0.167 rs2583321689471478G/AG = 0.950G = 0.9410.6810.840GA = 0.050A = 0.059rs20106231689472820C/T  C = 0.961   C = 0.978 0.4731.810T T = 0.039 T = 0.022rs1872831689473131G/AG = 0.955G = 0.9730.4701.690AA = 0.045A = 0.027rs2583301689473369T/G T = 0.583 T = 0.6260.2441.199GG = 0.416G = 0.373rs2583281689474109C/T  C = 0.870   C = 0.906 0.3401.430T T = 0.130 T = 0.094rs4599201689474622T/C T = 0.723 T = 0.6870.2540.841T  C = 0.276   C = 0.312 rs1662971689474980G/AG = 0.920G = 0.9080.5740.864GA = 0.079A = 0.091rs2583181689475845T/C T = 0.204 T = 0.2500.1161.300C  C = 0.795   C = 0.749 rs2583171689476033C/T  C = 0.978   C = 0.981 0.8531.123T T = 0.022 T = 0.019rs1718051689477409G/AG = 0.979G = 0.9540.2390.450GA = 0.021A = 0.046rs20819841689477721C/T  C = 0.874   C = 0.870 0.8300.958C T = 0.125 T = 0.129rs4645861689477883C/T  C = 0.354   C = 0.157 ˜0.0010.340C T = 0.645 T = 0.842rs4603191689477888G/TG = 0.166G = 0.2130.1031.362T T = 0.833 T = 0.786rs4477351689478144A/GA = 0.699A = 0.6600.2240.833AG = 0.300G = 0.339rs23770581689478626T/C T = 0.780 T = 0.7960.6091.100C  C = 0.220   C = 0.204 rs6048681689479972G/AG = 0.118G = 0.1160.9120.976GA = 0.881A = 0.883rs21181931689483698C/T  C = 0.814 C =     T = 0.186T =   rs4670351689484080C/G  C = 0.647   C = 0.632 0.6530.937CG = 0.352G = 0.367rs21154011689484404C/T  C = 0.660   C = 0.660 0.9991.000C T = 0.339 T = 0.339rs4173231689490072T/G T = 0.634 T = 0.6410.8771.027GG = 0.365G = 0.358rs23772331689490697C/G  C = 0.891   C = 0.845 0.1100.668CG = 0.108G = 0.154AA1689493039G/CG = 0.077G = 0.0420.0520.535Gds000175  C = 0.922   C = 0.957 rs97461141689495122C/G  C = 0.953   C = 0.948 0.7630.897CG = 0.046G = 0.051rs37517001689495989G/AG = 0.974G = 0.9530.5240.544GA = 0.025A = 0.046rs22779051689496296A/GA = 0.919A = 0.8860.1600.687AG = 0.080G = 0.113rs3978911689496826C/G  C = 0.695 C =    G = 0.305G =     rs37946381689497231G/TG = 0.924G = 0.9010.4890.751G T = 0.075 T = 0.098rs37946371689497365G/AG =     G =     A =     A =     rs2583241689498050C/A  C = 0.897   C = 0.891 0.7770.934CA = 0.102A = 0.108rs2583231689498489C/G  C = 0.823   C = 0.779 0.2250.760CG = 0.176G = 0.220rs20758801689499011A/GA = 0.623A = 0.6950.0271.380GG = 0.376G = 0.304rs2583221689499698C/T  C = 0.909   C = 0.950 0.0901.903T T = 0.090 T = 0.049rs2583211689500268C/TC =    C =    T =   T =   rs1647441689500563A/CA =     A =     C =    C =    rs38338291689500576A/—A = 0.103A = 0.1400.1231.411— = 0.896  — = 0.859  AB1689500994T/C T = 0.526 T = 0.4650.0640.783Tds000276  C = 0.473   C = 0.534 rs1647431689501226A/GA = 0.161A = 0.2440.0361.681GG = 0.838G = 0.755rs1647421689501520C/T  C = 0.652   C = 0.700 0.1281.247T T = 0.347 T = 0.299rs1872821689502768T/C T = 0.060 T = 0.0360.2260.588T  C = 0.939   C = 0.963 rs4655071689504287C/G  C = 0.194   C = 0.151 0.1120.735CG = 0.805G = 0.848rs21629431689504447G/AG = 0.796G = 0.8330.1721.281AA = 0.203A = 0.166AC1689504754G/CG = 0.238G = 0.1750.1050.681Gds000173  C = 0.761   C = 0.824 rs19464821689506205T/C T = 0.900 T = 0.8920.7140.915T  C = 0.099   C = 0.107 rs84041689506442C/T  C = 0.227   C = 0.141 0.0060.563C T = 0.772 T = 0.858rs10458141689506710G/AG = 0.659G = 0.6640.8641.025AA = 0.340A = 0.335rs42473531689507009T/C T = 0.376 T = 0.4180.3411.193C  C = 0.623   C = 0.581 rs4627691689507058A/GA = 0.665A = 0.6870.5401.106GG = 0.334G = 0.312rs38036901689507154C/G  C = 0.668   C = 0.702 0.3261.167GG = 0.331G = 0.297rs37516961689507414A/TA =     A =     T =   T =   rs99728641689508992A/GA = 0.048A = 0.0500.8721.058GG = 0.951G = 0.949rs105672261689509757TTT/—TTT = 0.916  TTT = 0.930  0.4301.233— = 0.083  — = 0.069  rs4174141689509943A/GA = 0.297A = 0.2730.886AG = 0.702G = 0.726rs1546611689510366G/CG = 0.045G = 0.0810.1481.838C  C = 0.954   C = 0.918 rs47857041689516165G/AG = 0.058G = 0.0560.8990.950GA = 0.942A = 0.944rs47855841689516179C/T  C = 0.875   C = 0.860 0.5770.880C T = 0.125 T = 0.140rs47855851689516200C/T  C = 0.797   C = 0.801 0.8821.030T T = 0.203 T = 0.199rs47857051689516209T/C T = 0.513 T = 0.4820.3830.880T  C = 0.487   C = 0.518 rs47855861689516211G/AG =     G =     A =     A =     rs47855871689516414A/GA = 0.758A = 0.7140.1340.795AG = 0.241G = 0.285rs20169681689516701C/G  C = 0.655   C = 0.639 0.6490.931CG = 0.344G = 0.360rs37516931689518191C/T  C = 0.377   C = 0.367 0.8160.960C T = 0.623 T = 0.633rs38096461689530650A/GA = 0.779A = 0.7460.2820.831AG = 0.220G = 0.253rs47855901689533962G/CG = 0.657G = 0.6210.3330.856G  C = 0.342   C = 0.378 rs20991051689537350A/GA = 0.439A = 0.3860.1170.802AG = 0.560G = 0.613rs47857081689540364C/T  C = 0.690   C = 0.640 0.1170.799C T = 0.309 T = 0.359rs47857101689540551T/C T = 0.553 T = 0.5980.2511.203C  C = 0.446   C = 0.401 rs47855911689541308C/T  C = 0.716   C = 0.755 0.2001.225T T = 0.283 T = 0.244rs47855921689541310C/G  C = 0.031   C = 0.010 0.0370.268CG = 0.969G = 0.990rs47855931689541422C/T  C = 0.396   C = 0.355 0.2180.843C T = 0.603 T = 0.644rs47857111689541741C/G  C = 0.977   C = 0.983 0.7301.376GG = 0.023G = 0.017rs47857121689541749G/CG = 0.239G = 0.2230.5840.914G  C = 0.760   C = 0.776 rs47857131689541893G/CG = 0.581G = 0.5270.1130.804G  C = 0.418   C = 0.472 rs47857141689542199C/A  C = 0.718   C = 0.631 0.0110.673CA = 0.281A = 0.368rs47855941689542311C/G  C = 0.708   C = 0.753 0.2101.256GG = 0.291G = 0.246rs38036891689543853A/GA = 0.681A = 0.6230.0690.776AG = 0.318G = 0.376rs47857151689543930G/AG = 0.631G = 0.6420.7381.047AA = 0.368A = 0.357rs37642581689544809C/G  C = 0.903   C = 0.910 0.7601.077GG = 0.096G = 0.089rs47857161689544922T/C T = 0.859 T = 0.956˜0.0013.627C  C = 0.140   C = 0.043 rs47857171689544928G/AG = 0.600G = 0.5710.3670.887GA = 0.399A = 0.428rs20770011689545885C/T  C = 0.412   C = 0.460 0.1601.217T T = 0.587 T = 0.539rs23770491689546763A/CA = 0.772A = 0.7920.5411.127C  C = 0.227   C = 0.207 rs20035221689547081C/G  C = 0.387   C = 0.345 0.2240.833CG = 0.612G = 0.654rs12301689548650C/T  C = 0.645   C = 0.666 0.5271.098T T = 0.354 T = 0.333rs18003591689549056A/GA = 0.705A = 0.7760.0151.448GG = 0.294G = 0.223rs10616461689549772T/C T = 0.387 T = 0.3550.3080.869T  C = 0.612   C = 0.644 rs18003581689553114C/T  C = 0.199   C = 0.199 0.9871.003C T = 0.800 T = 0.800rs22863921689553344C/T  C = 0.266   C = 0.270 0.8811.023T T = 0.733 T = 0.729rs20749041689555341T/C T = 0.914 T = 0.9190.8071.063C  C = 0.085   C = 0.080 rs20749031689555458A/GA = 0.921A = 0.9060.5070.829AG = 0.078G = 0.093









TABLE 25










Males























Melanoma



Chromo-
Chromosome

Male
Male
M
Odds
Associated


dbSNP rs#
some
Position
Alleles
Case AF
Control AF
p-Value
Ratio
Allele


















rs460879
16
89456684
T/C
 T = 0.532
T =   










  C = 0.468 

C =    


rs460984
16
89456969
T/C
T =   
T =   






C =    
C =    


rs2437957
16
89459443
C/T

  C = 0.541 


  C = 0.512 

0.430
0.890
C






 T = 0.458
 T = 0.487


rs3815949
16
89459711
C/G

  C = 0.015 


  C = 0.002 

0.207
0.108
C






G = 0.985
G = 0.998


rs2437956
16
89460046
G/A
G = 0.780
G = 0.723
0.081
0.737
G






A = 0.219
A = 0.276


rs2437955
16
89460070
G/T
G = 0.886
G = 0.825

0.037

0.605
G






 T = 0.113
 T = 0.174


rs2434872
16
89460710
C/G

  C = 0.544 


  C = 0.585 

0.352
1.183
G






G = 0.455
G = 0.414


rs467357
16
89461192
G/A
G = 0.654
G = 0.650
0.908
0.982
G






A = 0.345
A = 0.349


rs258337
16
89464505
A/T
A = 0.577
A = 0.654

0.028

1.385
T






 T = 0.422
 T = 0.345


rs258336
16
89464626
T/G
 T = 0.588
 T = 0.522
0.068
0.765
T






G = 0.411
G = 0.477


rs258335
16
89464739
A/G
A = 0.777
A = 0.791
0.709
1.085
G






G = 0.222
G = 0.208


rs164752
16
89466185
G/A
G = 0.101
G = 0.054

0.036

0.511
G






A = 0.898
A = 0.945


rs154663
16
89469830
T/C
 T = 0.172
 T = 0.207
0.270
1.252
C







  C = 0.827 


  C = 0.792 



rs164753
16
89471390
T/C
 T = 0.902
 T = 0.808

˜0.001

0.454
T







  C = 0.097 


  C = 0.191 



rs258332
16
89471478
G/A
G = 0.952
G = 0.933
0.383
0.700
G






A = 0.048
A = 0.067


rs2010623
16
89472820
C/T

  C = 0.971 


  C = 0.952 

0.535
0.590
C






 T = 0.029
 T = 0.048


rs187283
16
89473131
G/A
G = 0.967
G = 0.971
0.904
1.130
A






A = 0.033
A = 0.029


rs258330
16
89473369
T/G
 T = 0.632
 T = 0.606
0.456
0.895
T






G = 0.367
G = 0.393


rs258328
16
89474109
C/T

  C = 0.855 


  C = 0.782 

0.062
0.610
C






 T = 0.145
 T = 0.218


rs459920
16
89474622
T/C
 T = 0.718
 T = 0.686
0.317
0.857
T







  C = 0.281 


  C = 0.313 



rs166297
16
89474980
G/A
G = 0.944
G = 0.916
0.171
0.643
G






A = 0.055
A = 0.083


rs258318
16
89475845
T/C
 T = 0.149
 T = 0.223

0.027

1.633
C







  C = 0.850 


  C = 0.776 



rs258317
16
89476033
C/T

  C = 0.974 


  C = 0.978 

0.820
1.176
T






 T = 0.025
 T = 0.021


rs171805
16
89477409
G/A
G =     
G = 0.964






A =     
A = 0.036


rs2081984
16
89477721
C/T

  C = 0.860 


  C = 0.845 

0.552
0.887
C






 T = 0.139
 T = 0.154


rs464586
16
89477883
C/T

  C = 0.304 


  C = 0.132 


˜0.001

0.350
C






 T = 0.695
 T = 0.867


rs460319
16
89477888
G/T
G = 0.156
G = 0.230

0.010

1.617
T






 T = 0.843
 T = 0.769


rs447735
16
89478144
A/G
A = 0.686
A = 0.654
0.327
0.864
A






G = 0.313
G = 0.345


rs2377058
16
89478626
T/C
 T = 0.840
 T = 0.840
0.609
1.000
T







  C = 0.160 


  C = 0.160 



rs604868
16
89479972
G/A
G = 0.103
G = 0.109
0.791
1.063
A






A = 0.896
A = 0.890


rs2118193
16
89483698
C/T
C =    
C =    






T =   
T =   


rs467035
16
89484080
C/G

  C = 0.662 


  C = 0.620 

0.254
0.834
C






G = 0.337
G = 0.379


rs2115401
16
89484404
C/T

  C = 0.672 


  C = 0.651 

0.564
0.909
C






 T = 0.327
 T = 0.348


rs417323
16
89490072
T/G
 T = 0.625
 T = 0.663
0.444
1.183
G






G = 0.374
G = 0.336


rs2377233
16
89490697
C/G

  C = 0.899 


  C = 0.833 


0.038

0.562
C






G = 0.100
G = 0.166


AA
16
89493039
G/C
G = 0.049
G = 0.065
0.405
1.335
C







  C = 0.950 


  C = 0.934 



rs9746114
16
89495122
C/G

  C = 0.971 


  C = 0.942 

0.166
0.471
C






G = 0.028
G = 0.057


rs3751700
16
89495989
G/A
G = 0.987
G = 0.941
0.142
0.212
G






A = 0.013
A = 0.059


rs2277905
16
89496296
A/G
A = 0.955
A = 0.934
0.332
0.663
A






G = 0.044
G = 0.065


rs397891
16
89496826
C/G

  C = 0.608 

C =    






G = 0.392
G =     


rs3794638
16
89497231
G/T
G =     
G =     






T =   
T =   


rs3794637
16
89497365
G/A
G = 0.874
G = 0.832
0.189
0.716
G






A = 0.125
A = 0.167


rs258324
16
89498050
C/A

  C = 0.916 


  C = 0.893 

0.334
0.770
C






A = 0.083
A = 0.106


rs258323
16
89498489
C/G

  C = 0.816 


  C = 0.785 

0.300
0.824
C






G = 0.183
G = 0.214


rs2075880
16
89499011
A/G
A = 0.596
A = 0.707

0.001

1.631
G






G = 0.403
G = 0.292


rs258322
16
89499698
C/T

  C = 0.908 


  C = 0.954 


0.014

2.128
T






 T = 0.091
 T = 0.045


rs258321
16
89500268
C/T

  C = 0.606 


  C = 0.735 


0.003

1.801
T






 T = 0.393
 T = 0.264


rs164744
16
89500563
A/C
A = 0.715
A = 0.573

˜0.001

0.536
A







  C = 0.284 


  C = 0.426 



rs3833829
16
89500576
A/—
A = 0.103
A = 0.143
0.152
1.451







— = 0.896  
— = 0.856  


AB
16
89500994
T/C
 T = 0.492
 T = 0.425
0.078
0.764
T







  C = 0.507 


  C = 0.574 



rs164743
16
89501226
A/G
A =     
A =     






G =     
G =     


rs164742
16
89501520
C/T

  C = 0.678 


  C = 0.760 


0.015

1.501
T






 T = 0.321
 T = 0.239


rs187282
16
89502768
T/C
 T = 0.066
 T = 0.048
0.625
0.713
T







  C = 0.933 


  C = 0.951 



rs465507
16
89504287
C/G

  C = 0.218 


  C = 0.171 

0.106
0.739
C






G = 0.781
G = 0.828


rs2162943
16
89504447
G/A
G = 0.822
G = 0.797
0.470
0.851
G






A = 0.177
A = 0.202


AC
16
89504754
G/C
G = 0.253
G = 0.219
0.325
0.826
G







  C = 0.746 


  C = 0.780 



rs1946482
16
89506205
T/C
 T = 0.896
 T = 0.878
0.512
0.836
T







  C = 0.103 


  C = 0.121 



rs8404
16
89506442
C/T

  C = 0.195 


  C = 0.121 


0.017

0.567
C






 T = 0.804
 T = 0.878


rs1045814
16
89506710
G/A
G = 0.637
G = 0.675
0.328
1.183
A






A = 0.362
A = 0.324


rs4247353
16
89507009
T/C
T =   
T =   






C =    
C =    


rs462769
16
89507058
A/G
A = 0.663
A = 0.719
0.103
1.300
G






G = 0.336
G = 0.280


rs3803690
16
89507154
C/G

  C = 0.687 


  C = 0.690 

0.931
1.016
G






G = 0.312
G = 0.309


rs3751696
16
89507414
A/T
A = 0.725
A = 0.698
0.480
0.877
A






 T = 0.274
 T = 0.301


rs9972864
16
89508992
A/G
A = 0.047
A = 0.049
0.930
1.038
G






G = 0.952
G = 0.950


rs10567226
16
89509757
TTT/—

TTT = 0.925  


TTT = 0.936  

0.609
1.185







— = 0.074  
— = 0.063  


rs417414
16
89509943
A/G
A = 0.287
A = 0.235

˜0.001

0.766
A






G = 0.712
G = 0.764


rs154661
16
89510366
G/C
G = 0.025
G = 0.017
0.728
0.655
G







  C = 0.975 


  C = 0.983 



rs4785704
16
89516165
G/A
G = 0.049
G = 0.045
0.855
0.920
G






A = 0.951
A = 0.955


rs4785584
16
89516179
c/t

  C = 0.894 


  C = 0.869 

0.336
0.780
C






 T = 0.106
 T = 0.131


rs4785585
16
89516200
c/t

  C = 0.810 


  C = 0.811 

0.977
1.010
T






 T = 0.190
 T = 0.189


rs4785705
16
89516209
T/C
 T = 0.500
 T = 0.481
0.621
0.930
T







  C = 0.500 


  C = 0.519 



rs4785586
16
89516211
G/A
G = 0.804
G = 0.852
0.163
1.408
A






A = 0.195
A = 0.147


rs4785587
16
89516414
A/G
A = 0.742
A = 0.689
0.097
0.770
A






G = 0.257
G = 0.310


rs2016968
16
89516701
C/G

  C = 0.667 


  C = 0.635 

0.376
0.865
C






G = 0.332
G = 0.364


rs3751693
16
89518191
C/T

  C = 0.390 


  C = 0.382 

0.853
0.970
C






 T = 0.610
 T = 0.618


rs3809646
16
89530650
A/G
A = 0.786
A = 0.739
0.197
0.769
A






G = 0.213
G = 0.260


rs4785590
16
89533962
G/C
G = 0.643
G = 0.605
0.280
0.852
G







  C = 0.356 


  C = 0.394 



rs2099105
16
89537350
A/G
A = 0.420
A = 0.365
0.112
0.794
A






G = 0.579
G = 0.634


rs4785708
16
89540364
C/T

  C = 0.674 


  C = 0.600 


0.025

0.725
C






 T = 0.325
 T = 0.399


rs4785710
16
89540551
T/C
 T = 0.572
 T = 0.620
0.175
1.223
C







  C = 0.427 


  C = 0.379 



rs4785591
16
89541308
C/T

  C = 0.767 


  C = 0.768 

0.954
1.010
T






 T = 0.232
 T = 0.231


rs4785592
16
89541310
C/G

  C = 0.022 


  C = 0.003 

0.090
0.154
C






G = 0.978
G = 0.997


rs4785593
16
89541422
C/T

  C = 0.381 


  C = 0.340 

0.221
0.839
C






 T = 0.618
 T = 0.659


rs4785711
16
89541741
C/G

  C = 0.982 


  C = 0.990 

0.527
1.721
G






G = 0.018
G = 0.010


rs4785712
16
89541749
G/C
G = 0.233
G = 0.186
0.129
0.756
G







  C = 0.766 


  C = 0.813 



rs4785713
16
89541893
G/C
G = 0.548
G = 0.483
0.061
0.772
G







  C = 0.451 


  C = 0.516 



rs4785714
16
89542199
C/A

  C = 0.659 


  C = 0.599 

0.070
0.772
C






A = 0.340
A = 0.400


rs4785594
16
89542311
C/G

  C = 0.729 


  C = 0.752 

0.491
1.129
G






G = 0.270
G = 0.247


rs3803689
16
89543853
A/G
A = 0.637
A = 0.614
0.526
0.906
A






G = 0.362
G = 0.385


rs4785715
16
89543930
G/A
G = 0.641
G = 0.658
0.625
1.074
A






A = 0.358
A = 0.341


rs3764258
16
89544809
C/G

  C = 0.917 


  C = 0.912 

0.804
0.936
C






G = 0.082
G = 0.087


rs4785716
16
89544922
T/C
T =   
T =   






C =    
C =    


rs4785717
16
89544928
G/A
G = 0.599
G = 0.539
0.075
0.780
G






A = 0.400
A = 0.460


rs2077001
16
89545885
C/T

  C = 0.461 


  C = 0.474 

0.703
1.056
T






 T = 0.538
 T = 0.525


rs2377049
16
89546763
A/C
A = 0.777
A = 0.800
0.467
1.149
C







  C = 0.222 


  C = 0.199 



rs2003522
16
89547081
C/G

  C = 0.355 


  C = 0.315 

0.309
0.837
C






G = 0.644
G = 0.684


rs1230
16
89548650
C/T

  C = 0.663 


  C = 0.672 

0.774
1.043
T






 T = 0.336
 T = 0.327


rs1800359
16
89549056
A/G
A = 0.755
A = 0.805
0.085
1.340
G






G = 0.244
G = 0.194


rs1061646
16
89549772
T/C
 T = 0.375
 T = 0.332
0.261
0.830
T







  C = 0.624 


  C = 0.667 



rs1800358
16
89553114
C/T

  C = 0.204 


  C = 0.176 

0.362
0.835
C






 T = 0.795
 T = 0.823


rs2286392
16
89553344
C/T

  C = 0.268 


  C = 0.286 

0.582
1.094
T






 T = 0.731
 T = 0.713


rs2074904
16
89555341
T/C
 T = 0.931
 T = 0.919
0.537
0.839
T







  C = 0.068 


  C = 0.080 



rs2074903
16
89555458
A/G
A = 0.915
A = 0.914
0.943
0.981
A






G = 0.084
G = 0.085









Allelotyping results were considered significant with a calculated p-value of less than or equal to 0.05 for allelotype results. These values are indicated in bold. The assay failed for those SNPs in which the allele frequency is blank. The combined allelotyping p-values for males and females were plotted in FIG. 3A and separately for females and males in FIGS. 3B and 3C, respectively. The position of each SNP on the chromosome is presented on the x-axis. The y-axis gives the negative logarithm (base 10) of the p-value comparing the estimated allele frequency in the case group to that of the control group. The minor allele frequency of the control group for each SNP designated by an X or other symbol on the graphs in FIGS. 3A, 3B and 3C can be determined by consulting Table 24 or 25. By proceeding down the Table from top to bottom and across the graphs from left to right the allele frequency associated with each symbol shown can be determined.


To aid the interpretation, multiple lines have been added to the graph. The broken horizontal lines are drawn at two common significance levels, 0.05 and 0.01. The vertical broken lines are drawn every 20 kb to assist in the interpretation of distances between SNPs. Two other lines are drawn to expose linear trends in the association of SNPs to the disease. The light gray line (or generally bottom-most curve) is a nonlinear smoother through the data points on the graph using a local polynomial regression method (W. S. Cleveland, E. Grosse and W. M. Shyu (1992) Local regression models. Chapter 8 of Statistical Models in S eds J. M. Chambers and T. J. Hastie, Wadsworth & Brooks/Cole.). The black line (or generally top-most curve, e.g., see peak in left-most graph just to the left of position 92150000) provides a local test for excess statistical significance to identify regions of association. This was created by use of a 10 kb sliding window with 1 kb step sizes. Within each window, a chi-square goodness of fit test was applied to compare the proportion of SNPs that were significant at a test wise level of 0.01, to the proportion that would be expected by chance alone (0.05 for the methods used here). Resulting p-values that were less than 10−8 were truncated at that value.


Finally, the exons and introns of the genes in the covered region are plotted below each graph at the appropriate chromosomal positions. The gene boundary is indicated by the broken horizontal line. The exon positions are shown as thick, unbroken bars. An arrow is place at the 3′ end of each gene to show the direction of transcription.


Example 8
PCLO Proximal SNPs

It has been discovered that a polymorphic variation (rs1044639) in the untranslated region of a gene encoding presynaptic cytomatrix protein (PCLO) is associated with the occurrence of melanoma. See Table 3. Subsequently, seventy-six allelic variants located within or nearby the target gene were identified and subsequently allelotyped in melanoma case and control sample sets as described in Examples 1 and 2. Several SNPs, designated by SNP_ID “AD”, “AB”, and “AF” in column 1 of Table 26, are novel and were identified through SNP discovery methods using dye terminator sequencing. Methods of diedeoxynucleotide chain termination chemistry derived sequencing are described by Sanger et al. (Proc. Nat. Acad. Sci. USA 74, 5463-5467, 1997). Briefly, terminators are labeled with fluorescent dyes for automated detection. Fluorescein is measured by an argon ion laser in the sequencing instrument, which transfers light energy to a rhodamine acceptor dye. Each acceptor dye emits light at a characteristic wavelength for detection, identifying the nucleotide that terminated extension of the DNA fragment. Novel SNPs are identified when sequence from multiple samples are compared on one or more strands. The forward and reverse primers used to identify AD are CGTGCCTGATGGGATATACA and TTGGCTCAAAGGCAGCTAAT, respectively. The forward and reverse primers used to identify AE are TGCAACAACAAAGAGAACCA and TTCTCAGAGCCTTGTAGACTTCC, respectively. The forward and reverse primers used to identify AF are GTGAACTCATGCAGCAAGGA and GGTGGAACCTTTGCCAGTTA, respectively. The polymorphic variants are set forth in Table 26. The chromosome position provided in column four of Table 26 is based on Genome “Build 34” of NCBI's GenBank and dbSNP build “119”.

TABLE 26AlleleChromo-Position inChromosomeAllelePresent inIUPACdbSNP rs#someSEQ ID NO: 6PositionVariantsSEQ ID NO: 6Coders2158220722082023020T/CTYrs102446571,36582024165G/AARrs473248375,79882028598T/CCYrs473248578,87782031677C/TTYrs1859176717,30982040109C/TCYrs1859177717,44682040246C/TCYrs2074401718,83782041637A/GARrs4299948719,15982041959T/GGKrs4348435720,90282043702G/AARrs4348436720,92382043723G/CCSrs4413718721,06682043866G/CGSrs985199721,14782043947G/AGRrs7791413729,00882051808T/CTYrs2522830732,35482055154G/AGRrs2214414733,36382056163T/CCYrs2189246733,62382056423A/GGRrs5885312733,66982056469CTTAAGAA/—TNrs3064840733,67282056472AAGAACTT/—TNrs2051791733,98982056789C/TTYrs6467914734,59682057396C/TTYrs2214415734,73382057533G/AARrs2189247736,49682059296C/ACMrs3064869736,66982059469—/ACACNrs2522831737,35182060151T/CTYrs2715147737,65682060456T/CCYrs3064870738,05582060855—/TAAGGNrs2715148739,28682062086A/CAMrs2522832739,61582062415A/GARrs1044639740,80982063609G/TTKrs2715149742,06382064863A/TAWrs2522833742,95982065759C/AAMrs2247523743,65582066455C/GCSrs2371214743,92382066723T/CTYrs2189248744,55682067356T/CTYrs2715150745,14682067946A/GARrs2715151745,91482068714C/GCSrs2371215746,74582069545T/ATWrs2715152746,91782069717G/AGRrs2715153747,10982069909T/CCYrs972346749,78682072586C/TCYrs2715154750,11782072917G/TGKrs2522834750,19782072997T/CCYrs2023847750,54482073344G/TTKrs2715155751,30582074105A/GARrs2715156752,08582074885A/GGRrs10715043753,52382076323—/GGNAD753,84382076643C/GCSrs2301722754,48582077285A/CAMrs2257207757,43582080235A/TAWrs2715157757,62582080425A/GARrs2715158758,85782081657G/CGSrs2715159760,65982083459A/GARrs2522835760,74782083547A/TTWrs2522836762,30482085104C/ACMAE763,62482086424T/GTKrs2522837764,63382087433A/GARrs2522838764,80882087608A/TTWAF765,03982087839AT/—ANrs2522839766,89982089699G/AARrs2522840767,66682090466G/TTKrs2522841768,44582091245T/GGKrs10555175768,57782091377—/TTATTNrs10592258768,58182091381—/TATTATTTNrs2522842768,91382091713G/TGKrs6956848768,95082091750G/TTKrs2522843768,97382091773C/ACMrs2522844769,34382092143T/CCYrs2522845771,16082093960T/GTKrs2715161771,53282094332C/TCYrs6972720771,92682094726C/TTYrs4377905783,56282106362A/GARrs4579453783,97282106772T/CTYrs4260846784,08782106887C/ACMrs1986481785,93482108734A/TAWrs4628206786,59482109394A/TTWrs2888017789,45482112254A/GGR


Assay for Verifying and Allelotyping SNPs


The methods used to verify and allelotype the seventy-six proximal SNPs of Table 26 are the same methods described in Examples 1 and 2 herein. The primers and probes used in these assays are provided in Table 27 and Table 28, respectively.

TABLE 27dbSNP rs #Forward PCR primerReverse PCR primerrs2158220GGGCAGGAGAGAGATTCTATCCACAGCCCTTATTACAGTCrs1024465AGAGATTGTGTTAAGGCATCCAAGCAAAACTCTTCTAAAGrs4732483CCCACCAAGACTGTAAAAGCCAGGAAACAACAGATGCTGGrs4732485AAGTTCTTCCTGACAGCTTCAGGGAGTAGTGCAAGATAGGrs1859176GCTTATAGTGATTCATGCCCGGACAAGTTGTTTTTCACTGrs1859177CTACCAAAAGGGCACTTGACGATTAGTTATTGAGCAGAGGGrs2074401CCAGCATTTGTCAGATGAGGGGTTTAGGTTTCAATTCTGGrs4299948GAGAGAGGTTGGAATGACACGAGGGAGACTCCATCTCAAArs4348435CTAAGGCTACTACGAGCATGAAGGGTTAACAGAGTGCCTGrs4348436TTTTGGTTGAATTCTAAGGCAAGGGTTAACAGAGTGCCTGrs4413718TGTAGCTTTTTCTCTAGCCGGAGGACTGTAAGCTTAGAATGrs985199TTCCATCTTTTTAGAACGCCGAGAAAAAGCTACATGGCACrs7791413CCACTCTTTCCTTTACTTGGTAAGATACATCTTTGGGGCCrs2522830AATGCCAAAAGGATATTTGCTGTATGTAAAAATGATGACCrs2214414TTTCAGGGATCTGCAAGGTCAAAACTCCACTGTACACTCGrs2189246GACAGTCACCTTAACAATTACGAAACAAGAAGCTTAGAGACrs5885312GAGTTGTGAAGTTCTGAGTGTCAGCTTTCTTGAAGAGATTCrs3064840CTAGAGTTGTGAAGTTCTGAGCAGCTTTCTTGAAGAGATTCrs2051791GTCATTGGCAATGAGGATGCCAACTACCTAATTCAAATGTCrs6467914GAAAATTCATGTGTTTTTGGCGTATACTGAACAAGACCATGrs2214415TTCCACTTTTCATCCACATGTCTTCCCCCAGATCATGTTGrs2189247TCAAACAAAAATAGCTCAGGGAAGCATTCCTAAAACCTTGrs3064869ACCAAACCAACTAGCATGGGGCCTAAACAACTCTCCGTAGrs2522831GTACAGAGTGTTTTGTCATCCTGCTCACGCATAAAACAGGGrs2715147GATCCATGTATATTTCCAGGCCTAGTTTTTCTCTCTGCACTTrs3064870CTCTCACCTTTTCTTCCATCGGGAAGTGTTTGATTACTGTGrs2715148TACCAGTTGGAATGTGGATCCAAGAGAGAACTTTCACCATGrs2522832AATGCCATGCCCACAAAAGCAGCACACAGCAAGAACATCCrs1044639CTCCTTTGTTTCCACCATCCCGCAAACAAAAAGGACACACrs2715149AAACTGAGCTACTGCCTCTGGTTCAGTATATCTAGGTAAGGrs2522833TACCACCTTGGAGTGTTATCTTCCTTGGTTTATATCCTTGrs2247523GGGACAAAATGGAAAAGAATGGGTAGTCATATCCCTATTTCrs2371214TTGATCCTAGGCAGGAGTACGAAGGACTTCAGTAAGTCACrs2189248TACACTTACTATGTACTTGTAGATTAAAAATCACAAGATCrs2715150ACCATAGCAGCAAATAAGGGACACTGGAGGTGACAGTTTGrs2715151TAATTTTTGCCTACAGTTACGGCATTGTGGTGCTAATCTCrs2371215TAAGTTGAATGCACAGTGACTAGAACACCTACAAGCATTTCrs2715152CATCTTTTTCTCAGTACACTCGCCCCATAAACAATAATTTGGrs2715153AATGCAGATGTCCCAAGTTCATAAATTTTTGGTTTATGTCrs972346GGTGAGAAAACTTGAAGCTCCCTATGTTTGGATCCTGGTCrs2715154GCGTAATTGTTCCACAACACCATCGTGAACAAGTTAGGCCrs2522834TGGAGAATCAAACTCATTACTGGCCAGATACATTTAGAAArs2023847CAGCCTGACTTTATGCTCACCAAAATCTGTGGAGTTGAAGGrs2715155GAAGGCTTCTTTGGTGTATCGTTTAGTAAAGGAAAACCAGrs2715156ATTGATCAGAGGGTGGGAAGATAAGAGTCCTGTTAGCTAGrs10715043AGGGACCAAGTTCAAACACCCTGCACTCCAGCCTCAAAAAADTATACACCTGCACTGGACATGATTTAAGTTCTCGAGTCTTCrs2301722AGGCAGCTAATAAGCTACCCTTCCAAGGCCTTTGCTTGTCrs2257207GTACCGAAAGGTATCTATAGGGCTGTCAGCACTGAAATGTArs2715157GCACAATTGTGGCTGAATACGAAGAAAAAAATGCTTAACTCrs2715158AAGCAGAAGTGGAGTGTGAGAGGTCATCAGGGTAGTACTGrs2715159CCACAAAAGACCCTAATAGCAGTAGCATAATTCCTCTGGCrs2522835CCATTCCATATGTCTGTCTGAGCCAGAGGAATTATGCTACrs2522836ACAGAGTCTCACTCTGTCCCTCACTTGAATATGGGAGGCGAEGAGACATTTTTCAAGTTCCTCCTCTTCTGAGTACAAAACTGrs2522837AAACATAGGAAAAAGTGATGCATGATGAGATAGATTTAACrs2522838CCTGTGAAAGTAATGCTTGGCTAAAAAGAATCAAGTACGCAFTGCAGAGGCAGATGTAATTGGCTACAATGCCCTAGATTTTGrs2522839TACACCACTGCACTCCAGTCCCACAATTATGTTGTTGAACCrs2522840TGTGCTCCTGCAGAAATGTGGGAGGAAGAGTACATTAATArs2522841CTGGAAAATATTACCCATGTGTCTGTGTACTTTCATACCTGrs10555175ATAGTGCAAGACCCTGTCTCTGAAAGTACACAGATGAGACrs10592258ATAGTGCAAGACCCTGTCTCTGAAAGTACACAGATGAGACrs2522842AATCCTATCATCGATGCTACCCATGCCTGGCCTTTAATAGrs6956848CTTTTTCAGTGTTTAGTAGCCGAGTTTTGCTGAATATGTAGCrs2522843TCGATGATAGGATTTGTTGCTACTAAGTTCTTGAATTACCrs2522844CCAATTGTTTTCTAGTAGACTTGGTAGGCACTACACAATTCrs2522845ATGTACACCAGTGGTTTGCCAAACCAGTAGTGGATCAGCCrs2715161TAGCTCCTACATATGGAGTGAACAGGGCTGGAATTGGAGGrs6972720TCTCCAGATCCTATTCTATGCACAACTCATGTACATCAGGrs4377905AATGCAAACTGTCTAAGCACTATGGCTGTATCAATATCACrs4579453TCTCAGCTCACTGCACTGTCGACTGTAATCCCAGCTACTCrs4260846AGAGCAAGATGCCGTCTCAGGGAAATATACCATATAAATGrs1986481AAGCATGACATTATGTTTCCAGTACCCAGGTGTTCTTAGCrs4628206GCGTTAGGTACACATGACACCTTCACAGAAGAGAGTTTCCrs2888017CACTAGTCATCAGGGAAATCCATCTCCTTGCCAGCATCTC












TABLE 28













Term












dbSNP rs #
Extend Primer
Mix
















rs2158220
CTGAGGCCTGCCCCTGA
ACT








rs1024465
TTAAGGCATCTCTGTATATACTAA
ACG







rs4732483
CAGATGCTGGAGAGGAT
ACT







rs4732485
GACAGCTTCAGATATTTCA
ACG







rs1859176
CATGCCCAATTCATATAGATA
ACG







rs1859177
TCTGTCTTGTTCCCATC
ACG







rs2074401
CAGATGAGGTTTCTGAAG
ACT







rs4299948
AAGGACTATAGGGCTACTA
ACT







rs4348435
CATGCCTACCTGTTAATATG
ACG







rs4348436
GAATTCTAAGGCTACTACGA
ACT







rs4413718
AAGAAGTGGTCTACTGTC
ACT







rs985199
GAACGCCTTATCTAAGCA
ACG







rs7791413
CTTGGTACTTAGTAATACACTTA
ACT







rs2522830
AGGATATTTGCAAACGTG
ACG







rs2214414
TGCAAGGTCAAAATATTCTT
ACT







rs2189246
ATCAAGTTTTCAGCTTTCTTGAA
ACT







rs5885312
GTGAAGTTCTGAGTGAAACAAG
CGT







rs3064840
GTGAAGTTCTGAGTGAAACAAG
CGT







rs2051791
GATGCTTTAAAACAAAAACTACA
ACG







rs6467914
ATGTGTTTTTGGCTTTTTAGTGA
ACG







rs2214415
CCATAATATTTCACCCATTCT
ACG







rs2189247
AAAATAGCTCAGGTTTTTCA
CGT







rs3064869
CATGGGGAAAACAACAACAAC
CGT







rs2522831
ACATTAAGTGGATGAGGT
ACT







rs2715147
TATTTCCAGGCACTTCA
ACT







rs3064870
TTCTTCATGTTATTCACGTTCTT
CGT







rs2715148
GAATGTGGATCAGTGTTT
ACT







rs2522832
AGACAATCTCTAAAAATTAA
ACT







rs1044639
CATCCATCCAACCTGGCTC
CGT







rs2715149
CCTCTGTGGGATAAAACA
CGT







rs2522833
GAGTGTTATCGAGGTGAG
CGT







rs2247523
TGAGTAAGTTGCAATTACAAA
ACT







rs2371214
CAGGAGTACTCTAGATTAGT
ACT







rs2189248
CAAATAATCATGAAATTGGTAGC
ACT







rs2715150
AAATAAGGGAAAGGAAGTC
ACT







rs2715151
TTGCCTACAGTTACTTATCT
ACT







rs2371215
GTAGTTTTCTTTGCCCT
CGT







rs2715152
CAATGTTTTAGTTTTGCTTTTC
ACG







rs2715153
TTTGGTTCATCAATTGTAAAATA
ACT







rs972346
GAAGCTCAGAAAAATTAAGG
ACG







rs2715154
TTCCACAACACTCACTT
CGT







rs2522834
CATTTTAACTTTTGTCCAATCA
ACT







rs2023847
TAAGAATCCCCCTGTTT
CGT







rs2715155
GGTGTATCTACTTTCATAAAATT
ACT







rs2715156
AGGGTGGGAAGAAAAAA
ACT







rs10715043
TCAAACACCTATTATCTTTTTTTTT
ACT







AD
GCACTGGACATCCAGAAGAC
ACT







rs2301722
GGTCTTTCTTTTAATCACAC
ACT







rs2257207
CGAAAGGTATCTATAGGTTTAATA
CGT







rs2715157
AACAAATATTTGAGAAAACTGC
ACT







rs2715158
TGTGCAGAAGGTTGACA
ACT







rs2715159
CCTAATAGCCAAAGCAATTTT
ACT







rs2522835
CATATGTCTGTCTGCTTTTA
CGT







rs2522836
CAGTGGTGTGATCTCTG
CGT







AE
TTCAAGTTCCTCATAATAACTAC
ACT







rs2522837
CAATGGATTTAACAAATTTGTAG
ACT







rs2522838
ATGCTTGGAGCCATTTC
CGT







AF
AATTGTAGATGAAGATGTAGACA
CGT







rs2522839
GACAGAGTGGGACCTGT
ACG







rs2522840
TGCAGAAATGTGACAATG
CGT







rs252284
TTATTACCCATGTGAATTATGATAT
ACT







rs10555175
GACCCTGTCTCAAAAAATAAATAA
CGT







rs10592258
GACCCTGTCTCAAAAAATAAATAA
CGT







rs2522842
GATGCTACATATTCAGCAAAA
CGT







rs6956848
CCTATTAAGCAAAAGACTTTGC
CGT







rs2522843
CAAAGTCTTTTGCTTAATAGG
CGT







rs2522844
ACACCATAAAATGTTAGACATAA
ACT







rs2522845
CTGCTGGCTTCCTACTT
ACT







rs2715161
CTACATATGGAGTGAGAACA
ACG







rs6972720
ATGGGTACCCTTAGACACAC
ACG







rs4377905
AACTGTCTAAGCACTCTAAA
ACT







rs4579453
TCTGCCTCCCTGGTTCA
ACT







rs4260846
ACAAACAAACAAACAAAAAAC
CGT







rs1986481
TTTTGCCAACTATTATCCC
CGT







rs4628206
CATGACACAGACCACAT
CGT







rs2888017
GAAATCCAAATTAAAACCACA
ACT











Genetic Analysis


Allelotyping results are shown for female (F) and male (M) cases and controls in Table 29 and Table 30, respectively. Allele frequency is noted in the fourth and fifth columns for melanoma pools and control pools, respectively.

TABLE 29FemalesMelanomaChromo-ChromosomeFemaleFemaleFOddsAssociateddbSNP rs#somePositionAllelesCase AFControl AFp-ValueRatioAllelers2158220782023020T/CT =T =C =C =rs1024465782024165G/AG = 0.845G = 0.8450.9870.997AA = 0.154A = 0.154rs4732483782028598T/CT = 0.112T = 0.1080.8660.960TC = 0.888C = 0.892rs4732485782031677C/TC = 0.050C = 0.0470.8680.946CT = 0.949T = 0.952rs1859176782040109C/TC = 0.647C = 0.761˜0.0011.738TT = 0.352T = 0.238rs1859177782040246C/TC = 0.979C = 0.9810.6551.291TT = 0.021T = 0.019rs2074401782041637A/GA = 0.620A = 0.5570.0960.771AG = 0.379G = 0.442rs4299948782041959T/GT =T =G =G =rs4348435782043702G/AG = 0.735G = 0.7420.8281.033AA = 0.264A = 0.257rs4348436782043723G/CG = 0.813G = 0.8210.7821.050CC = 0.186C = 0.178rs4413718782043866G/CG = 0.960G = 0.9680.5961.267CC = 0.039C = 0.031rs985199782043947G/AG = 0.244G = 0.2440.9801.004GA = 0.755A = 0.755rs7791413782051808T/CT = 0.697T = 0.6500.1950.809TC = 0.302C = 0.349rs2522830782055154G/AG = 0.630G = 0.5690.0820.775GA = 0.369A = 0.430rs2214414782056163T/CT = 0.582T = 0.6510.0491.340CC = 0.417C = 0.348rs2189246782056423A/GA = 0.160A = 0.1260.1970.756AG = 0.839G = 0.873rs5885312782056469CTTAAGAA/—CTTAAGAA = 0.070CTTAAGAA = 0.1010.1371.504— = 0.929— = 0.898rs3064840782056472AAGAACTT/—AAGAACTT = 0.042AAGAACTT = 0.0500.6241.196— = 0.957— = 0.949rs2051791782056789C/TC = 0.449C = 0.5410.0071.445TT = 0.550T = 0.458rs6467914782057396C/TC = 0.121C = 0.2040.0011.852TT = 0.878T = 0.795rs2214415782057533G/AG = 0.576G = 0.6110.3871.156AA = 0.423A = 0.388rs2189247782059296C/AC = 0.689C = 0.6070.0270.697CA = 0.310A = 0.392rs3064869782059469—/ACA— = 0.555— = 0.6350.0151.393AACA = 0.444ACA = 0.364rs2522831782060151T/CT =T =C =C =rs2715147782060456T/CT = 0.597T = 0.6560.0901.285CC = 0.402C = 0.343rs3064870782060855—/TAAG— = 0.279— = 0.2520.3980.872TAAG = 0.720TAAG = 0.747rs2715148782062086A/CA = 0.558A = 0.4630.0040.681AC = 0.441C = 0.536rs2522832782062415A/GA =A =G =G =rs1044639782063609G/TG = 0.413G = 0.4850.0261.339TT = 0.586T = 0.514rs2715149782064863A/TA = 0.927A = 0.9540.2561.648TT = 0.072T = 0.045rs2522833782065759C/AC = 0.384C = 0.523˜0.0011.762AA = 0.615A = 0.476rs2247523782066455C/GC = 0.633C = 0.5750.0790.783CG = 0.366G = 0.424rs2371214782066723T/CT = 0.637T = 0.5690.0420.752TC = 0.362C = 0.430rs2189248782067356T/CT = 0.276T = 0.2740.9450.990TC = 0.723C = 0.725rs2715150782067946A/GA = 0.787A = 0.7490.2050.811AG = 0.212G = 0.250rs2715151782068714C/GC = 0.632C = 0.5460.0100.699CG = 0.367G = 0.453rs2371215782069545T/AT = 0.626T = 0.5360.0080.688TA = 0.373A = 0.463rs2715152782069717G/AG = 0.680G = 0.6490.3220.868GA = 0.319A = 0.350rs2715153782069909T/CT = 0.549T = 0.6040.1681.254CC = 0.450C = 0.395rs972346782072586C/TC = 0.949C = 0.9570.6311.182TT = 0.050T = 0.042rs2715154782072917G/TG = 0.652G = 0.5580.0060.673GT = 0.347T = 0.441rs2522834782072997T/CT = 0.424T = 0.4240.9950.999CC = 0.575C = 0.575rs2023847782073344G/TG = 0.464G = 0.3570.0010.641GT = 0.535T = 0.642rs2715155782074105A/GA = 0.674A = 0.5920.0260.701AG = 0.325G = 0.407rs2715156782074885A/GA = 0.496A = 0.5560.0801.276GG = 0.503G = 0.443rs10715043782076323—/G— = 0.501— = 0.5580.0821.258GG = 0.498G = 0.441AD782076643C/GC = 0.973C = 0.9650.5820.773CG = 0.026G = 0.034rs2301722782077285A/CA = 0.966A = 0.9610.7290.864AC = 0.033C = 0.038rs2257207782080235A/TA = 0.612A = 0.5570.1230.798AT = 0.387T = 0.442rs2715157782080425A/GA = 0.614A = 0.5290.0090.707AG = 0.385G = 0.470rs2715158782081657G/CG = 0.742G = 0.6800.0480.740GC = 0.257C = 0.319rs2715159782083459A/GA = 0.726A = 0.619˜0.0010.613AG = 0.273G = 0.380rs2522835782083547A/TA = 0.542A = 0.685˜0.0011.840TT = 0.457T = 0.314rs2522836782085104C/AC = 0.532C = 0.5190.7140.948CA = 0.467A = 0.480AE782086424T/GT = 0.957T = 0.9910.0095.183GG = 0.043G = 0.009rs2522837782087433A/GA = 0.649A = 0.5640.0190.700AG = 0.350G = 0.435rs2522838782087608A/TA = 0.458A = 0.5120.1271.239TT = 0.541T = 0.487AF782087839AT/—AT = 0.668AT = 0.5920.0280.721A— = 0.331— = 0.407rs2522839782089699G/AG = 0.650G = 0.6760.5341.126AA = 0.349A = 0.323rs2522840782090466G/TG = 0.511G = 0.5550.2881.193TT = 0.488T = 0.444rs2522841782091245T/GT = 0.422T = 0.4880.0671.305GG = 0.577G = 0.511rs10555175782091377—/TTAT— = 0.623— = 0.7070.0181.460TTTAT = 0.376TTAT = 0.292rs10592258782091381—/TATTATT— = 0.618— = 0.6820.0731.326TTATTATT = 0.381TATTATT = 0.317rs2522842782091713G/TG = 0.648G = 0.5840.0620.764GT = 0.351T = 0.415rs6956848782091750G/TG =G =T =T =rs2522843782091773C/AC = 0.567C = 0.5690.9561.007AA = 0.432A = 0.430rs2522844782092143T/CT = 0.459T = 0.5170.1091.262CC = 0.540C = 0.482rs2522845782093960T/GT = 0.657T = 0.5920.0450.757TG = 0.342G = 0.407rs2715161782094332C/TC = 0.616C = 0.5580.0830.786CT = 0.383T = 0.441rs6972720782094726C/TC = 0.777C = 0.7790.9401.012TT = 0.222T = 0.220rs4377905782106362A/GA = 0.955A = 0.9600.7601.123GG = 0.044G = 0.039rs4579453782106772T/CT = 0.954T = 0.9110.0160.490TC = 0.045C = 0.088rs4260846782106887C/AC = 0.729C = 0.7090.5100.906CA = 0.270A = 0.290rs1986481782108734A/TA = 0.437A = 0.3750.0580.772AT = 0.562T = 0.624rs4628206782109394A/TA = 0.854A = 0.8460.7570.941AT = 0.145T = 0.153rs2888017782112254A/GA = 0.868A = 0.8650.9070.976AG = 0.131G = 0.134









TABLE 30










Males























Melanoma



Chromo-
Chromosome

Male
Male
M
Odds
Associated


dbSNP rs#
some
Position
Alleles
Case AF
Control AF
p-Value
Ratio
Allele


















rs2158220
7
82023020
T/C
T = 0.740
T = 0.682
0.076
0.753
T






C = 0.259
C = 0.317


rs1024465
7
82024165
G/A
G = 0.864
G = 0.878
0.622
1.127
A






A = 0.135
A = 0.121


rs4732483
7
82028598
T/C
T = 0.115
T = 0.111
0.903
0.970
T






C = 0.885
C = 0.889


rs4732485
7
82031677
C/T
C = 0.040
C = 0.030
0.572
0.741
C






T = 0.959
T = 0.969


rs1859176
7
82040109
C/T
C =
C =






T =
T =


rs1859177
7
82040246
C/T
C = 0.989
C = 0.985
0.743
0.716
C






T = 0.011
T = 0.015


rs2074401
7
82041637
A/G
A = 0.633
A = 0.574
0.095
0.781
A






G = 0.366
G = 0.425


rs4299948
7
82041959
T/G
T = 0.089
T = 0.337
1.332
5.184
G






G = 0.910
G = 0.662


rs4348435
7
82043702
G/A
G = 0.760
G = 0.772
0.669
1.073
A






A = 0.239
A = 0.227


rs4348436
7
82043723
G/C
G = 0.831
G = 0.864
0.205
1.293
C






C = 0.168
C = 0.135


rs4413718
7
82043866
G/C
G = 0.959
G = 0.972
0.375
1.486
C






C = 0.040
C = 0.027


rs985199
7
82043947
G/A
G = 0.224
G = 0.222
0.949
0.989
G






A = 0.775
A = 0.777


rs7791413
7
82051808
T/C
T = 0.725
T = 0.698
0.396
0.877
T






C = 0.274
C = 0.301


rs2522830
7
82055154
G/A
G = 0.659
G = 0.574

0.015

0.696
G






A = 0.340
A = 0.425


rs2214414
7
82056163
T/C
T = 0.556
T = 0.632
0.054
1.369
C






C = 0.443
C = 0.367


rs2189246
7
82056423
A/G
A =
A =






G =
G =


rs5885312
7
82056469
CTTAAGAA/—
CTTAAGAA = 0.057
CTTAAGAA = 0.061
0.803
1.083







— = 0.942
— = 0.938


rs3064840
7
82056472
AAGAACTT/—
AAGAACTT = 0.043
AAGAACTT = 0.039
0.859
0.919
A






— = 0.956
— = 0.960


rs2051791
7
82056789
C/T
C = 0.469
C = 0.562

0.025

1.455
T






T = 0.530
T = 0.437


rs6467914
7
82057396
C/T
C = 0.149
C = 0.164
0.569
1.121
T






T = 0.850
T = 0.835


rs2214415
7
82057533
G/A
G = 0.623
G = 0.621
0.976
0.994
G






A = 0.376
A = 0.378


rs2189247
7
82059296
C/A
C = 0.653
C = 0.626
0.408
0.886
C






A = 0.346
A = 0.373


rs3064869
7
82059469
—/ACA
— = 0.553
— = 0.605
0.127
1.238
A






ACA = 0.446
ACA = 0.394


rs2522831
7
82060151
T/C
T = 0.714
T = 0.699
0.709
0.932
T






C = 0.285
C = 0.300


rs2715147
7
82060456
T/C
T = 0.572
T = 0.611
0.338
1.176
C






C = 0.427
C = 0.388


rs3064870
7
82060855
—/TAAG
— = 0.231
— = 0.207
0.420
0.869







TAAG = 0.768
TAAG = 0.792


rs2715148
7
82062086
A/C
A = 0.557
A = 0.502
0.141
0.802
A






C = 0.442
C = 0.497


rs2522832
7
82062415
A/G
A = 0.662
A = 0.628
0.335
0.861
A






G = 0.337
G = 0.371


rs1044639
7
82063609
G/T
G = 0.429
G = 0.488
0.090
1.273
T






T = 0.570
T = 0.511


rs2715149
7
82064863
A/T
A = 0.907
A = 0.940
0.097
1.620
T






T = 0.092
T = 0.059


rs2522833
7
82065759
C/A
C = 0.396
C = 0.503

0.003

1.543
A






A = 0.603
A = 0.496


rs2247523
7
82066455
C/G
C = 0.646
C = 0.582
0.061
0.764
C






G = 0.353
G = 0.417


rs2371214
7
82066723
T/C
T = 0.619
T = 0.592
0.471
0.893
T






C = 0.380
C = 0.407


rs2189248
7
82067356
T/C
T = 0.246
T = 0.238
0.821
0.958
T






C = 0.753
C = 0.761


rs2715150
7
82067946
A/G
A = 0.802
A = 0.778
0.441
0.868
A






G = 0.197
G = 0.221


rs2715151
7
82068714
C/G
C = 0.631
C = 0.597
0.414
0.864
C






G = 0.368
G = 0.402


rs2371215
7
82069545
T/A
T = 0.620
T = 0.544

0.025

0.731
T






A = 0.379
A = 0.455


rs2715152
7
82069717
G/A
G = 0.718
G = 0.688
0.450
0.866
G






A = 0.281
A = 0.311


rs2715153
7
82069909
T/C
T = 0.491
T = 0.595

0.016

1.523
C






C = 0.508
C = 0.404


rs972346
7
82072586
C/T
C = 0.959
C = 0.970
0.485
1.362
T






T = 0.040
T = 0.029


rs2715154
7
82072917
G/T
G = 0.632
G = 0.573
0.084
0.784
G






T = 0.367
T = 0.426


rs2522834
7
82072997
T/C
T = 0.386
T = 0.428
0.222
1.189
C






C = 0.613
C = 0.571


rs2023847
7
82073344
G/T
G = 0.467
G = 0.400
0.052
0.761
G






T = 0.532
T = 0.599


rs2715155
7
82074105
A/G
A =
A =






G =
G =


rs2715156
7
82074885
A/G
A = 0.411
A = 0.533

0.006

1.634
G






G = 0.588
G = 0.466


rs10715043
7
82076323
—/G
— = 0.474
— = 0.533
0.092
1.263
G






G = 0.525
G = 0.466


AD
7
82076643
C/G
C = 0.967
C = 0.977
0.506
1.410
G






G = 0.032
G = 0.022


rs2301722
7
82077285
A/C
A = 0.965
A = 0.976
0.418
1.487
C






C = 0.034
C = 0.023


rs2257207
7
82080235
A/T
A = 0.622
A = 0.582
0.234
0.843
A






T = 0.377
T = 0.417


rs2715157
7
82080425
A/G
A =
A =






G =
G =


rs2715158
7
82081657
G/C
G = 0.748
G = 0.682
0.091
0.724
G






C = 0.251
C = 0.317


rs2715159
7
82083459
A/G
A = 0.733
A = 0.634

0.002

0.632
A






G = 0.266
G = 0.365


rs2522835
7
82083547
A/T
A = 0.534
A = 0.651

˜0.001

1.630
T






T = 0.465
T = 0.348


rs2522836
7
82085104
C/A
C = 0.570
C = 0.544
0.506
0.899
C






A = 0.429
A = 0.455


AE
7
82086424
T/G
T = 0.940
T = 0.967
0.217
1.886
G






G = 0.059
G = 0.032


rs2522837
7
82087433
A/G
A = 0.639
A = 0.606
0.341
0.869
A






G = 0.360
G = 0.393


rs2522838
7
82087608
A/T
A = 0.451
A = 0.517
0.078
1.301
T






T = 0.548
T = 0.482


AF
7
82087839
AT/—
AT = 0.676
AT = 0.592

0.028

0.721
A






— = 0.323
— = 0.407


rs2522839
7
82089699
G/A
G =
G =






A =
A =


rs2522840
7
82090466
G/T
G = 0.488
G = 0.550
0.101
1.284
T






T = 0.511
T = 0.449


rs2522841
7
82091245
T/G
T = 0.411
T = 0.462
0.181
1.230
G






G = 0.588
G = 0.537


rs10555175
7
82091377
—/TTAT
— = 0.620
— = 0.679
0.089
1.294
T






TTAT = 0.379
TTAT = 0.320


rs10592258
7
82091381
—/TATTATT
— = 0.613
— = 0.670
0.090
1.283
T






TATTATT = 0.386
TATTATT = 0.329


rs2522842
7
82091713
G/T
G = 0.647
G = 0.603
0.190
0.829
G






T = 0.352
T = 0.396


rs6956848
7
82091750
G/T
G = 0.035
G = 0.150

˜0.001

4.818
T






T = 0.964
T = 0.849


rs2522843
7
82091773
C/A
C = 0.576
C = 0.542
0.352
0.872
C






A = 0.423
A = 0.457


rs2522844
7
82092143
T/C
T = 0.458
T = 0.501
0.219
1.187
C






C = 0.541
C = 0.498


rs2522845
7
82093960
T/G
T = 0.677
T = 0.604

0.047

0.725
T






G = 0.322
G = 0.395


rs2715161
7
82094332
C/T
C = 0.666
C = 0.591
0.051
0.725
C






T = 0.333
T = 0.408


rs6972720
7
82094726
C/T
C = 0.796
C = 0.791
0.839
0.966
C






T = 0.203
T = 0.208


rs4377905
7
82106362
A/G
A = 0.955
A = 0.975
0.180
1.828
G






G = 0.044
G = 0.024


rs4579453
7
82106772
T/C
T = 0.959
T = 0.899

0.006

0.382
T






C = 0.040
C = 0.100


rs4260846
7
82106887
C/A
C = 0.726
C = 0.707
0.590
0.912
C






A = 0.273
A = 0.292


rs1986481
7
82108734
A/T
A = 0.485
A = 0.417

0.049

0.760
A






T = 0.514
T = 0.582


rs4628206
7
82109394
A/T
A = 0.863
A = 0.858
0.826
0.954
A






T = 0.136
T = 0.141


rs2888017
7
82112254
A/G
A = 0.902
A = 0.895
0.767
0.927
A






G = 0.097
G = 0.104









Allelotyping results were considered significant with a calculated p-value of less than or equal to 0.05 for allelotype results. These values are indicated in bold. The assay failed for those SNPs in which the allele frequency is blank. The combined allelotyping p-values for males and females were plotted in FIG. 4A and separately for females and males in FIGS. 4B and 4C, respectively. The position of each SNP on the chromosome is presented on the x-axis. The y-axis gives the negative logarithm (base 10) of the p-value comparing the estimated allele frequency in the case group to that of the control group. The minor allele frequency of the control group for each SNP designated by an X or other symbol on the graphs in FIGS. 4A, 4B and 4C can be determined by consulting Table 29 or 30. By proceeding down the Table from top to bottom and across the graphs from left to right the allele frequency associated with each symbol shown can be determined.


To aid the interpretation, multiple lines have been added to the graph. The broken horizontal lines are drawn at two common significance levels, 0.05 and 0.01. The vertical broken lines are drawn every 20 kb to assist in the interpretation of distances between SNPs. Two other lines are drawn to expose linear trends in the association of SNPs to the disease. The light gray line (or generally bottom-most curve) is a nonlinear smoother through the data points on the graph using a local polynomial regression method (W. S. Cleveland, E. Grosse and W. M. Shyu (1992) Local regression models. Chapter 8 of Statistical Models in S eds J. M. Chambers and T. J. Hastie, Wadsworth & Brooks/Cole.). The black line (or generally top-most curve, e.g., see peak in left-most graph just to the left of position 92150000) provides a local test for excess statistical significance to identify regions of association. This was created by use of a 10 kb sliding window with 1 kb step sizes. Within each window, a chi-square goodness of fit test was applied to compare the proportion of SNPs that were significant at a test wise level of 0.01, to the proportion that would be expected by chance alone (0.05 for the methods used here). Resulting p-values that were less than 10−8 were truncated at that value.


Finally, the exons and introns of the genes in the covered region are plotted below each graph at the appropriate chromosomal positions. The gene boundary is indicated by the broken horizontal line. The exon positions are shown as thick, unbroken bars. An arrow is place at the 3′ end of each gene to show the direction of transcription.


Example 9
FPGT/CARK Proximal SNPs

It has been discovered that a polymorphic variation (rs2034453) in an intron of a gene encoding cardiac troponin I interacting kinase (TNNI3K), also referred to as cardiac ankyrin repeat kinase (CARK), and near a gene encoding fucose-1-phosphate guanylyltransferase (FPGT) is associated with the occurrence of melanoma. See Table 3. Subsequently, ninety-two allelic variants located within or nearby the target genes were identified and subsequently allelotyped in melanoma case and control sample sets as described in Examples 1 and 2. Several SNPs, designated by SNP_ID “AG”, “AH”, and “AI” in column 1 of Table 20, are novel and were identified through SNP discovery methods using dye terminator sequencing. Methods of diedeoxynucleotide chain termination chemistry derived sequencing are described by Sanger et al. (Proc. Nat. Acad. Sci. USA 74, 5463-5467, 1997). Briefly, terminators are labeled with fluorescent dyes for automated detection. Fluorescein is measured by an argon ion laser in the sequencing instrument, which transfers light energy to a rhodamine acceptor dye. Each acceptor dye emits light at a characteristic wavelength for detection, identifying the nucleotide that terminated extension of the DNA fragment. Novel SNPs are identified when sequence from multiple samples are compared on one or more strands. The forward and reverse primers used to identify AG are GGGTGGCTCACTCTCTAGGAA and CCTAGCAGCTGCCATAGCC, respectively. The forward and reverse primers used to identify AH are AACTTGTCCACAACCCTCAGTT and TGAAGAGTTTGAATGTGAAGCTG, respectively. The forward and reverse primers used to identify AI are TGGCTTAGGAGAAAATGATCTTG and TTTTCTGTAACATTATACCAAGGAAC, respectively. The polymorphic variants are set forth in Table 31. The chromosome position provided in column four of Table 31 is based on Genome “Build 34” of NCBI's GenBank and dbSNP build “119”.

TABLE 31PositionAlleleChromo-in SEQChromosomeAllelePresent inIUPACdbSNP rs#someID NO: 5PositionVariantsSEQ ID NO: 5CodeAG120674033656G/TTKrs1412825129974033749G/CCSrs4422957163474034084C/TTYrs3835390199074034440AAAAG/—ANrs94479511,43374034883C/GGSrs79231011,97874035428G/AGRrs52673612,22774035677A/TAWrs980400612,99274036442T/CTYrs57736714,17174037621C/TARrs57575414,31374037763T/CGRrs57372114,58674038036G/TCMrs48791715,38174038831C/TGRrs79230715,48574038935G/ACYrs47635015,79274039242T/CCYrs942502316,39674039846T/CTYrs942502416,45874039908G/TTKrs57184818,00474041454A/GARrs48994118,80874042258A/CCMrs49162318,95974042409A/GGRrs95619,12574042575T/ATWAH19,26174042711G/AGRrs52080619,82674043276C/TCYrs522759110,06774043517T/CTYrs545664110,26274043712G/AARrs545721110,28474043734C/TTYrs536355110,66174044111G/AARrs553044111,05874044508G/TTKrs581353111,87174045321A/GARrs496008112,29174045741T/AAWrs7366854113,15074046600C/TCYrs560808114,71874048168C/ACMrs485929114,77074048220A/GARrs477134115,39974048849T/GTKrs505634116,13674049586G/AARrs542136116,34574049795G/AARrs545680116,72574050175T/CTYrs522042117,56774051017G/AGRrs575961117,69574051145C/TTYrs792321118,45374051903A/GARrs524306118,73874052188T/AAWrs495027118,74474052194A/CCMrs525358119,78974053239A/GARrs471496120,39174053841G/AGRrs504866121,72574055175C/TTYrs532396123,37774056827T/CCYrs533371123,49774056947G/AARrs567060124,84974058299A/GARrs4650251130,03374063483A/GGRrs3753183135,58474069034G/TTKrs518769136,39774069847A/GGRrs474215136,73674070186T/CCYAI137,59274071042T/CCYrs500203138,13674071586T/AAWrs532221138,43574071885T/ATWrs539534139,22274072672C/TTYrs531412139,27074072720G/AARrs9970514140,24374073693C/GGSrs568049140,98874074438C/TTYrs9727250144,06574077515T/CCYrs792323144,07974077529C/TCYrs3765651145,84774079297A/CCMrs3765654146,19674079646C/GCSrs481387146,93574080385C/TCYrs483259147,11474080564T/CCYrs2034453148,66674082116A/GARrs7536997149,43774082887G/AGRrs792324149,55674083006T/CTYrs6660350150,03674083486C/TCYrs492302150,62474084074G/CTSrs10493539152,06274085512A/GARrs496720155,19874088648G/AARrs570631156,34074089790T/ATWrs4593767159,57874093028G/AGRrs576802160,78174094231T/AAWrs518574167,52374100973A/GARrs524252169,08274102532C/TCYrs792327170,00674103456G/AARrs792328170,43774103887C/AAMrs2039407170,85574104305A/TAWrs503770171,93174105381T/CTYrs503904171,99074105440A/GARrs514012173,56674107016A/GARrs1412827174,08274107532A/GARrs473834179,51974112969G/AARrs480267180,20974113659A/GGRrs572180185,68474119134G/TGKrs1333029185,80774119257G/AGRrs485414185,92774119377A/GARrs1412823186,25174119701A/TTWrs2027013187,43874120888T/GAMrs548881195,03074128480C/TCYrs792329197,17574130625C/TAR


Assay for Verifying and Allelotyping SNPs


The methods used to verify and allelotype the ninety-two proximal SNPs of Table 31 are the same methods described in Examples 1 and 2 herein. The primers and probes used in these assays are provided in Table 32 and Table 33, respectively.

TABLE 32dbSNP rs #Forward PCR primerReverse PCR primerAGATTTGTCTGACTGTGTTGGGTTTGTCACCCAAACTGAACCrs1412825TTGGCTGGCGCCCGGAAAATCCTGAGACCGTTGCTAGAGrs4422957GTTTGCTGGACTGTCACTTCATTAGCTAGCTACTCCTGGGrs3835390TGAGGGAGTAACAGAACATGAGGGATTAAGTGTCATAGCCrs944795GAAGGGATCTTTGGGTTGTGCCAGCTTTGCAAATCAATTCrs792310ACTGACCATAAAGCGCGTACAAAGGAGTTACCCCTTGGAGrs526736CCTTGAAAAACTTCCAGAATGACTGATAAGGTCCAAAGGAGrs9804006CACTAGTTATCTTGGGCCAGAAAGTGCTGGGATTACAGGCrs577367TTTTTAAGCATTGTGGCCTCTACTAGACTTTTGCCAGTACrs575754GAAAAGTAGGAAGGACATGGCAAAGGTTTTCATGTAACCArs573721CTGGTTTGAAGCCTAAAAATGCCTGGTATCTAATTTAATTTTrs487917TCACCTAGTGATGTTGGAGCTACCAGCATCACCACTTGAGrs792307GGGACAACCTATGTATATGCAGAGCCAGACTCCATCTCAArs476350ATACCAACACTTTGGGAGGCGGTTTCACCGTGTTAGCCAGrs9425023TATATGCCACTTGTTACTGCTGGCAACAAGAAAATTTCTCrs9425024CCCAGGGTTTTATAACCCATGATTTGCAGTAACAAGTGGCrs571848CAGGGAACAAATTTTTCTAGCTCAAAAAGGCAAAGTGTACrs489941TTGTTTCTCAGCATCACCAGCTCCTATCAGTCCTTCTAACrs491623AGAAGCAGGCAGGAAAATGGCATAACTTGAGATTCTAGCCrs956AGTCTGTGCTACTTTATGTCCTCCTTGGGAAAAACTTTGGAHTGATGGCACTTCAGATTTGGCCAAAGTTTTTCCCAAGGAGrs520806GTCAGAAAGAGATTTCTCACATTCAGACAGGTCACTTCTGGrs522759CAAAGCAACCAGAATCCATGGCTCAATACTGTATTTTAACCrs545664CCCAGAAAACAGAATGTGCCGACCAAGTTCACATGTGAGCrs545721GACCAAGTTCACATGTGAGCCCCAGAAAACAGAATGTGCCrs536355ATAGCAGGCTTTTACACACCGACCTACATGGATAAGTTTGrs553044CAAGTCCAGACGGCATCATGCTGCCTTCTGTCACATATAGrs581353ATCACCTACCTGCACATCAGCAATCAGTTACTTGCATGCACrs496008ACCAACACATCATAGCTAAGGAGAAAACAGTGGATACTGGrs7366854GGATATTAGCCCTTCGTCAGCTACCATCAGAGTGAACAGGrs560808CAATGCCTTATTGTGTGCAGAGGCTTTCTGCAACTTTGCGrs485929ACGCAAAGTTGCAGAAAGCCTTCAATGCCTCATACACCCCrs477134CTGAATCATTGGAGCTAGGGCTGTGCTCTGATATGTATCCrs505634AGGAGTGAATGTGGATAGAGTTCTCCTCATCTTCCTCACCrs542136CTCTGTTATTGGAAAAAATTCAAGCCAGCAGGATTATGAAGrs545680AATGAAAGTACACTCTACAGCCCCCACAAATTCTGTAACGrs522042GAAGGAGTCTTGCTCTGTTGAGGAGAATGGCCTGAACCTGrs575961GCACAAAATATACCATCGTCGCACTGGACACCTGTTATTCrs792321GTTTCCCATGGCCTAAGATGCTTCTCTGGTCTCCTGATAGrs524306TTGGTTTGGGATAAAGGAGGAAGCCTCCCAAAAATATGCCrs495027CCTAATAATCTAAAGCCTCCCTTGGTTTGGGATAAAGGAGGrs525358ATCTCTTAGTTTGGTACGCGCACACTACACTTTGACTGGGrs471496TCATTTCCAAGAAATATGTGGTTTATTTACTTTGAACCTGrs504866TCTAAATTGTGTCCAGACTCTAAAAATCTTAGACAGTGGGrs532396TTCTAACTGTGGTGTTTCTGCAAGACTTGTAATCAAATCCrs533371GGAAATTGTACATGCAAAGGTTACTGGTACGGGAAGAATGrs567060ATAAACCAAGAGGCATGGTGAACAAGAGCGAAACCCTGTCrs4650251TAGAGCAATCAGAGAAGAGGGTGGAGTCTTTAGGTTTTTCCrs3753183AGCTTTGGCTTTCAAAGGTGTGTGGGAGAATTCAACTGAGrs518769GGAAGCCTTACCTCTGAAACCAAACAACTCTCTTTTTGAArs474215CAGTTCTGACCATGTGACTCATGGAAGGTGCATGAAAGAGAICCAGTACAAGTTTGGGTTGGGTCACTGCACTTGAACTTGGrs500203TCGAATGGGTTCTTCATTACGCAAAATGTAGACACAGATCrs532221CTTTGTATAAGTGTCTTCAGTATGTCTGAGGTTCTACGGGrs539534GCAATGACCAAATGACACATCCATGGGTGATTTTATTTGCCrs531412CCTTGTTACATCCATTGACGGCAATGACCAAATGACACATCrs9970514TCCTACCATACCCTGCTTACTCTTGCCTACACACCTTTACrs568049GATAAGGAAGCTGAGGAGAGCCATTAACATAAGAGGCCCCrs9727250AATGGCGTGAATCCAGGAGGTTTCAGACGGAGTCTCGCTCrs792323TTTTTCAGACGGAGTCTCGCAATGGCGTGAATCCAGGAGGrs3765651CCCAAAGGTTACTTTCTTTCTGGATCACAAAACTGCACACrs3765654GGCAAAAGAAGAGAAAATGGCTTCTAGTCAAACGTTTCTGrs481387AACCTGAAGTGAGCCTTAAGCCATTCAATAAGTCTGGCACrs483259CAGAACTGCACATGAGATTCTTTGAGTCTCTCCACTGTTGrs2034453GTGACTGGAAACTGAGAATGTTGCTGGACAATAGAAAGACrs7536997AGGAATGAACAACTCCGGACCGACCTTCGCAGTGAGTGTTrs792324TGCTTCTCACTCTTTGGGTCTGTGATCTTGCTGGCTTCAGrs6660350CTAGTAGATCCCACGCAGGGAGTCCCATCCACCACCCAArs492302TCTTATTTCTAATATCTCGGTTCCTCCATATGTGGAAAACrs10493539GAAGCACTTAAAGATTCCCCGGCATTTATCACTTTTCCACrs496720TGTGTCTAGCTCAGGGATTGTGGTCCGTTTTACAGAGAGCrs570631TTGCATTTTGTCCCATTTCCAAGACTGAAGTGGCAACAGGrs4593767ATTGATATATGAAGAAACAGGTCCAATCTGCTGTTTTCAACrs576802TCTACTTTTGCATTGCTTCCGGCAATGGATGACAATAGTArs518574GAGCTCCAGAGGAAAAATTGAGGTGGTGGTAAAAGAAGGGrs524252TGGGAGATCTATTCTACCTCTGGAAGAATGAGGCTTAGACrs792327TGGTCCGTTTTACAGAGAGCTGTGTCTAGCTCAGGGATTGrs792328TCAGCAGGCCATAGCTGCCTCATGCAAGAGCCCATGGCAGGrs2039407AATGACTATGTGCACTGGAGTAAGGTGGTTACAATTACTCrs503770AATGCATCTTTGGTCCTGACTTGAGATGCCATTGATAGGCrs503904GAAGTCAGGACCAAAGATGCCTAAGCTGCTTTCCCATTCCrs514012TCTTGTTCAAGCGGTAGAGGCTCAGATGGTGGAAAGTAGGrs1412827GCTTTGGATAGTCAATTCAGCCTCTGTATATCTAGCTACCrs473834TCTTGTGGGTATAAGCTGTGCTACCTTCACATCTGTAGTCrs480267CATCCAAATGTTTCAGTAGAGTTGGAGAGGAAACTAAAGCGrs572180AGCTTTTCTTGCTTATGCTCCAGGTCTTTGGCATAGACTCrs1333029GGGATTTCTCATATCAGGTTGGCCCTGGAAAATGATGGAAGrs485414GCACTGAATTTGCTTCCAACGTGTATATACATTATCTACATrs1412823GTATTCCTAATTAAGGGGAAGAAACCTCTGAGCTCACTATGrs2027013CTTCCCGTTTTTCTACCTGCGAGCCAATGAAAGAAGAAAGGrs548881GTAGTTCTATAAGGTTTTGCCCACCCTGGGAAACTATAAACrs792329CACCACGCCCAGCTAATTTTGTCAGGAGATCAAGACCATC












TABLE 33













Term












dbSNP rs #
Extend Primer
Mix
















AG
CTGTGTTGGGGCTGGGCC
CGT








rs1412825
GCGCCCGGAAAACCATGAT
ACT







rs4422957
CCGTTTACTTCTCATCTG
ACG







rs3835390
TGGAGGAATTATGGATTAAAAATT
ACT







rs944795
CCTAACACCAAGTGTCC
ACT







rs792310
TTGGCTCCAGCAGGATC
ACG







rs526736
TTCCAGAATGTTACTTAAAAGA
CGT







rs9804006
GGGCCAGTTACTTAACTCTTG
ACT







rs577367
GCATTGTGGCCTCTGGCTA
ACG







rs575754
GGACATGGTCTCAGAAT
ACT







rs573721
GAAGCCTAAAAATGTGTAAAA
CGT







rs487917
CCATTGTAAAGTCCTAGCA
ACG







rs792307
GTATATGCTGTCCATAATTGAG
ACG







rs476350
GGCAGGCAGATCACGAG
ACT







rs9425023
TCAGGAAAGAATAATGAAATCCA
ACT







rs9425024
CCCATAAGTTACAAATCAAAATAT
CGT







rs571848
AAAAGCAGTTTGATGTAGAG
ACT







rs489941
TAAAGTCAGTTCATCTGGTAATT
ACT







rs491623
ACAGGTGATAAACAGTTTC
ACT







rs956
GTCAATATAAAAATTGGTAAAC
CGT







AH
GAATGACAAAGACATAACATCCT
ACG







rs520806
AAGAGATTTCTCACAAATTGA
ACG







rs522759
CAGAATCCATGCATTGT
ACT







rs545664
GCTATTAGCCAGGCAGCAA
ACG







rs545721
GCTGCCTGGCTAATAGCTA
ACG







rs536355
CTGAGTATATGATGTAAG
ACG







rs553044
GTATGTAAACAGCAACTCTG
CGT







rs581353
GTCTAAAGATGTTCATCCC
ACT







rs496008
TGCACATGTAAATGATTTTTTT
CGT







rs7366854
CTTCGTCAGATGAGTAGGTTG
ACG







rs560808
GTGTGCAGTAATAACAATGAT
CGT







rs485929
AGAAAGCCTGCAAAGGA
ACT







rs477134
TGGAGCTAGGGAGGAAG
ACT







rs505634
ATGTGGATAGAGAAATTCAAA
ACG







rs542136
TTTTCTTCCTTGTAGCATC
ACG







rs545680
CACTCTACAGTGTGGGA
ACT







rs522042
TAGCTCACTCCAAGCTC
ACG







rs575961
CATCGTCATAGTAGAGATTC
ACG







rs792321
AAGATGAATGGACTCTCCTGT
ACT







rs524306
GGACATATTTACTTCTAGCTGAT
CGT







rs495027
CCTCCCAAAAATATGCCTTTG
ACT







rs525358
CGAGAGTGCTCGGTGCATA
ACT







rs471496
GAAATATGTGTTTTAACGAGAA
ACG







rs504866
GTGTCCAGACTCTTGATT
ACG







rs532396
CTGTGGTGTTTCTGATGCCT
ACT







rs533371
TGCAAAGGTGTTAGAAGCACCTC
ACG







rs567060
AGGCATGGTGCTTAGAT
ACT







rs4650251
GAAGAGGGAAAAAAGGACTTC
ACT







rs3753183
CGATGAGTTTGAAAATCCA
CGT







rs518769
CTGAGCAAAAATATATGGTGA
ACT







rs474215
GTGACTCTAATATCCAAGCTA
ACT







AI
AAGTTTCTTCCTTTCTCCAGG
ACT







rs500203
AAACATCTGCATTTTAAAC
CGT







rs532221
TGTCTTCAGTATTCACCTTA
CGT







rs539534
GACCAAATGACACATCTTTAAAA
ACG







rs531412
AACTATATTATACATGGGTGATTT
ACG







rs9970514
TCCTGATTTATTCATTGATTTTTG
ACT







rs568049
GAGGAGAGGATTATCCAGTC
ACG







rs9727250
TCGCGCCACTGCACTCCA
ACT







rs792323
AGGCTGGAGTGCAGTGG
ACG







rs3765651
TTCCTAGCCTTTACCTTT
ACT







rs3765654
AAAAGAAGAGAAAATGGAATTTCT
ACT







rs481387
AGGCGTATAGTAGGTGTTTAG
ACG







rs483259
CTCTTATTTAAACTTGATCTCTC
ACT







rs2034453
AAACTGAGAATGTTGATGGACA
ACT







rs7536997
AACAACTCCAGATGCGCCG
ACG







rs792324
GTCCGCACTGCCTTTAT
ACT







rs6660350
CACGCAGGGGCCGCAGG
ACG







rs492302
TCTCGGATCTTCCATCC
ACT







rs10493539
TCATCAAGCAGTTGCTTCACA
ACT







rs496720
GTAAATGCACCAATCAGCACC
ACG







rs570631
TCCCATTTCCATTATTTTTTTT
CGT







rs4593767
GAAGAAACAGAAAAATGTGC
ACG







rs576802
TTGCATTGCTTCCAAAATGATC
CGT







rs518574
AGAGGAAAAATTGCCTATG
ACT







rs524252
TTCTCAAGGATATAGCTGGAG
ACG







rs792327
ATGCACCAATCAGCACC
ACG







rs792328
GGGAAGCTCAGGCATGG
CGT







rs2039407
GTGCACTGGAGCAGTTCTG
CGT







rs503770
CTGCTAAGTCTGAGTCCCAT
ACT







rs503904
AAGATGCATTTATCAGATTGTATA
ACT







rs514012
GTAGAGGATTGATTAGAACTGA
ACT







rs1412827
TTCTAACAGTGTATTTAATCATCA
ACT







rs473834
CAGAAGAGTCTCTGGGGAG
ACG







rs480267
ATTTTGTTTACCAAGAAGCCTC
ACT







rs572180
CTTATGCTCTTCTACCTCA
CGT







rs1333029
GTTGATTTGGTTAGCAATAAT
ACG







rs485414
AACATCCTAGGTCCTCT
ACT







rs1412823
CCTAATTAAGGGGAAGAAGAAG
CGT







rs2027013
AAGAAGAAAGGTTGAAGAC
ACT







rs548881
GGTTTTGCCTAATATATTTTGATG
ACG







rs792329
GACCATCCTGGCTAACA
ACG










Genetic Analysis


Allelotyping results are shown for female (F) and male (M) cases and controls in Table 34 and Table 35, respectively. Allele frequency is noted in the fourth and fifth columns for melanoma pools and control pools, respectively.

TABLE 34FemalesMelanomaChromo-ChromosomeFemaleFemaleFOddsAssociateddbSNP rs#somePositionAllelesCase AFControl AFp-ValueRatioAlleleAG174033656G/TG = 0.167G = 0.1060.0270.593G T = 0.832 T = 0.893rs1412825174033749G/CG = 0.539G = 0.4990.2690.853G  C = 0.460   C = 0.500 rs4422957174034084C/T  C = 0.484   C = 0.427 0.0890.796C T = 0.515 T = 0.572rs3835390174034440AAAAG/—AAAAG = 0.580   AAAAG = 0.623   0.1991.199— = 0.419  — = 0.376  rs944795174034883C/G  C = 0.592   C = 0.534 0.0880.791CG = 0.407G = 0.465rs792310174035428G/AG = 0.508G = 0.5660.0831.265AA = 0.491A = 0.433rs526736174035677A/TA = 0.637A = 0.6530.6981.070T T = 0.362 T = 0.346rs9804006174036442T/C T = 0.748 T = 0.7000.1120.785T  C = 0.251   C = 0.299 rs577367174037621C/T  C = 0.540   C = 0.460 0.0250.727C T = 0.459 T = 0.539rs575754174037763T/C T = 0.923 T = 0.9380.4141.257C  C = 0.076   C = 0.061 rs573721174038036G/TG = 0.145G = 0.1510.8611.050T T = 0.854 T = 0.848rs487917174038831C/T  C = 0.567   C = 0.642 0.0201.369T T = 0.432 T = 0.357rs792307174038935G/AG = 0.530G = 0.6170.0021.431AG = 0.495G = 0.597rs792307174038935G/AA = 0.469A = 0.3820.0021.431AA = 0.504A = 0.402rs476350174039242T/C T = 0.036 T = 0.0420.7291.192C  C = 0.963   C = 0.957 rs9425023174039846T/C T = 0.595 T = 0.6260.3401.142C  C = 0.404   C = 0.373 rs9425024174039908G/TG = 0.955G = 0.9540.9760.989G T = 0.044 T = 0.045rs571848174041454A/GA = 0.555A = 0.6070.1161.237GG = 0.444G = 0.392rs489941174042258A/CA = 0.564A = 0.4650.0030.672A  C = 0.435   C = 0.534 rs491623174042409A/GA = 0.621A = 0.5280.0080.682AG = 0.378G = 0.471rs956174042575T/A T = 0.613 T = 0.6420.4001.129AA = 0.386A = 0.357AH174042711G/AG = 0.976G = 0.9670.5010.707GA = 0.023A = 0.032rs520806174043276C/T  C = 0.586   C = 0.643 0.0791.276T T = 0.413 T = 0.356rs522759174043517T/C T = 0.691 T = 0.7420.0881.287C  C = 0.308   C = 0.257 rs545664174043712G/AG = 0.562G = 0.5030.0740.789GA = 0.437A = 0.496rs545721174043734C/T  C = 0.510   C = 0.458 0.1720.813C T = 0.489 T = 0.541rs536355174044111G/AG = 0.534G = 0.4650.0490.758GA = 0.465A = 0.534rs553044174044508G/TG = 0.503G = 0.4870.6580.938G T = 0.496 T = 0.512rs581353174045321A/GA = 0.604A = 0.6430.2811.183GG = 0.395G = 0.356rs496008174045741T/A T = 0.864 T = 0.8110.0400.674TA = 0.135A = 0.188rs7366854174046600C/T  C = 0.840   C = 0.851 0.6771.089T T = 0.159 T = 0.148rs560808174048168C/A  C = 0.566   C = 0.640 0.0331.360AA = 0.433A = 0.359rs485929174048220A/GA = 0.716A = 0.7180.9431.011GG = 0.283G = 0.281rs477134174048849T/G T = 0.560 T = 0.6240.0561.306GG = 0.439G = 0.375rs505634174049586G/AG = 0.923G = 0.9450.2511.447AA = 0.076A = 0.054rs542136174049795G/AG = 0.625G = 0.5250.0070.661GA = 0.374A = 0.474rs545680174050175T/C T = 0.685 T = 0.7190.2761.175C  C = 0.314   C = 0.280 rs522042174051017G/AG = 0.699G =     A = 0.301A =     rs575961174051145C/T  C = 0.979   C = 0.948 0.1160.390C T = 0.020 T = 0.051rs792321174051903A/GA = 0.644A = 0.6750.3841.148GG = 0.355G = 0.324rs524306174052188T/A T = 0.559 T = 0.4680.0050.693TA = 0.440A = 0.531rs495027174052194A/CA = 0.585A = 0.4990.0110.707A  C = 0.414   C = 0.500 rs525358174053239A/GA = 0.150A = 0.1410.7310.930AG = 0.849G = 0.858rs471496174053841G/AG = 0.115G = 0.1230.7431.078AA = 0.884A = 0.876rs504866174055175C/T  C = 0.577   C = 0.465 0.0020.639C T = 0.422 T = 0.534rs532396174056827T/C T = 0.951 T = 0.9550.7901.102C  C = 0.048   C = 0.044 rs533371174056947G/AG = 0.528G = 0.4840.1840.840GA = 0.471A = 0.515rs567060174058299A/GA = 0.670A = 0.7070.2481.189GG = 0.329G = 0.292rs4650251174063483A/GA = 0.571A = 0.4730.0030.675AG = 0.428G = 0.526rs3753183174069034G/TG = 0.090G = 0.0530.0460.571G T = 0.909 T = 0.946rs518769174069847A/GA = 0.875A = 0.8750.9961.001AG = 0.124G = 0.124rs474215174070186T/C T = 0.556 T = 0.4520.0020.658T  C = 0.443   C = 0.547 AI174071042T/C T = 0.031 T = 0.0410.5511.349C  C = 0.968   C = 0.958 rs500203174071586T/A T = 0.507 T = 0.4250.0160.718TA = 0.492A = 0.574rs532221174071885T/A T = 0.725 T = 0.7830.0431.368AA = 0.274A = 0.216rs539534174072672C/TC =    C =    T =   T =   rs531412174072720G/AG = 0.682G = 0.7530.0341.423AA = 0.317A = 0.246rs9970514174073693C/G  C = 0.421   C = 0.474 0.1351.241GG = 0.578G = 0.525rs568049174074438C/T  C = 0.783   C = 0.781 0.9710.993C T = 0.216 T = 0.218rs9727250174077515T/C T = 0.074 T = 0.0570.3360.758T  C = 0.925   C = 0.942 rs792323174077529C/T  C = 0.815   C = 0.817 0.9331.014T T = 0.462 T = 0.436rs3765651174079297A/CA = 0.666A = 0.7100.1741.232C  C = 0.333   C = 0.289 rs3765654174079646C/G  C = 0.799   C = 0.686 ˜0.0010.551CG = 0.200G = 0.313rs481387174080385C/T  C = 0.517   C = 0.447 0.0480.755C T = 0.482 T = 0.552rs483259174080564T/C T = 0.580 T = 0.6650.0151.438C  C = 0.419   C = 0.334 rs2034453174082116A/GA = 0.208A = 0.1430.0150.634AG = 0.791G = 0.856rs7536997174082887G/AG =     G =     A =     A =     rs792324174083006T/C T = 0.067 T = 0.0530.4960.779T T = 0.038 T = 0.036rs792324174083006T/C  C = 0.932   C = 0.946 0.4960.779T  C = 0.961   C = 0.963 rs6660350174083486C/T  C = 0.191   C = 0.126 0.0100.607C T = 0.808 T = 0.873rs492302174084074G/CG =     G = 0.389C =      C = 0.611 rs10493539174085512A/GA = 0.375A = 0.3300.1750.821AG = 0.624G = 0.669rs496720174088648G/AG = 0.964G = 0.9770.3941.598AA = 0.035A = 0.022rs570631174089790T/A T = 0.375 T = 0.3920.6881.074AA = 0.624A = 0.607rs4593767174093028G/AG = 0.231G = 0.2700.1821.234AA = 0.768A = 0.729rs576802174094231T/A T = 0.860 T = 0.8460.5690.898TA = 0.139A = 0.153rs518574174100973A/GA = 0.972A = 0.9690.8610.920AG = 0.027G = 0.030rs524252174102532C/T  C = 0.828   C = 0.826 0.9250.983C T = 0.171 T = 0.173rs792327174103456G/AG = 0.957G = 0.9600.8931.070AA = 0.043A = 0.040rs792328174103887C/AC =      C = 0.337 A =     A = 0.663rs2039407174104305A/TA = 0.930A = 0.9620.0491.924T T = 0.069 T = 0.037rs503770174105381T/C T = 0.853 T = 0.8160.1370.764T  C = 0.146   C = 0.183 rs503904174105440A/GA = 0.838A = 0.8090.2690.820AG = 0.161G = 0.190rs514012174107016A/GA = 0.860A = 0.8110.0620.698AG = 0.139G = 0.188rs1412827174107532A/GA = 0.385A = 0.3270.0910.777AG = 0.614G = 0.672rs473834174112969G/AG = 0.142G = 0.1910.0501.425AA = 0.857A = 0.808rs480267174113659A/GA = 0.317A = 0.3560.2731.190GG = 0.682G = 0.643rs572180174119134G/TG = 0.868G = 0.8380.2020.786G T = 0.131 T = 0.161rs1333029174119257G/AG = 0.945G = 0.9710.0881.948AA = 0.054A = 0.028rs485414174119377A/GA = 0.911A = 0.9060.8240.947AG = 0.088G = 0.093rs1412823174119701A/TA = 0.774A = 0.7790.8761.030T T = 0.225 T = 0.220rs2027013174120888T/G T = 0.148 T = 0.0750.0050.470TG = 0.852G = 0.925rs548881174128480C/T  C = 0.898   C = 0.862 0.1760.709C T = 0.101 T = 0.137rs792329174130625C/T  C = 0.732   C = 0.744 0.6881.070T T = 0.268 T = 0.256









TABLE 35










Males























Melanoma



Chromo-
Chromosome

Male
Male
M
Odds
Associated


dbSNP rs#
some
Position
Alleles
Case AF
Control AF
p-Value
Ratio
Allele


















AG
1
74033656
G/T
G = 0.136
G = 0.103
0.178
0.730
G






 T = 0.863
 T = 0.896


rs1412825
1
74033749
G/C
G = 0.520
G = 0.518
0.947
0.990
G







  C = 0.479 


  C = 0.481 



rs4422957
1
74034084
C/T

  C = 0.462 


  C = 0.450 

0.739
0.955
C






 T = 0.537
 T = 0.549


rs3835390
1
74034440
AAAAG/—
AAAAG = 0.618   
AAAAG = 0.599   
0.588
0.924
A






— = 0.381  
— = 0.400  


rs944795
1
74034883
C/G

  C = 0.553 


  C = 0.540 

0.707
0.948
C






G = 0.446
G = 0.459


rs792310
1
74035428
G/A
G = 0.548
G = 0.564
0.630
1.069
A






A = 0.451
A = 0.435


rs526736
1
74035677
A/T
A =     
A =     






T =   
T =   


rs9804006
1
74036442
T/C
 T = 0.724
 T = 0.691
0.306
0.853
T







  C = 0.275 


  C = 0.308 



rs577367
1
74037621
C/T
C =    
C =    






T =   
T =   


rs575754
1
74037763
T/C
 T = 0.929
 T = 0.946
0.343
1.346
C







  C = 0.070 


  C = 0.053 



rs573721
1
74038036
G/T
G = 0.094
G = 0.137
0.210
1.525
T






 T = 0.905
 T = 0.862


rs487917
1
74038831
C/T

  C = 0.629 


  C = 0.684 

0.159
1.282
T






 T = 0.370
 T = 0.315


rs792307
1
74038935
G/A
G = 0.591
G = 0.654
0.286
1.306
A






G = 0.543
G = 0.583


rs792307
1
74038935
G/A
A = 0.408
A = 0.345
0.286
1.306
A






A = 0.456
A = 0.416


rs476350
1
74039242
T/C
 T = 0.052
 T = 0.047
0.767
0.886
T







  C = 0.947 


  C = 0.952 



rs9425023
1
74039846
T/C
 T = 0.614
 T = 0.594
0.577
0.919
T







  C = 0.385 


  C = 0.405 



rs9425024
1
74039908
G/T
G = 0.953
G = 0.981
0.138
2.541
T






 T = 0.047
 T = 0.019


rs571848
1
74041454
A/G
A = 0.600
A = 0.602
0.944
1.010
G






G = 0.399
G = 0.397


rs489941
1
74042258
A/C
A = 0.500
A = 0.481
0.601
0.928
A







  C = 0.499 


  C = 0.518 



rs491623
1
74042409
A/G
A = 0.577
A = 0.565
0.771
0.954
A






G = 0.422
G = 0.434


rs956
1
74042575
T/A
 T = 0.624
 T = 0.627
0.939
1.011
A






A = 0.375
A = 0.372


AH
1
74042711
G/A
G = 0.966
G = 0.961
0.762
0.878
G






A = 0.033
A = 0.038


rs520806
1
74043276
C/T

  C = 0.631 


  C = 0.664 

0.324
1.155
T






 T = 0.368
 T = 0.335


rs522759
1
74043517
T/C
 T = 0.724
 T = 0.752
0.361
1.154
C







  C = 0.275 


  C = 0.247 



rs545664
1
74043712
G/A
G = 0.534
G = 0.524
0.794
0.961
G






A = 0.465
A = 0.475


rs545721
1
74043734
C/T

  C = 0.534 


  C = 0.479 

0.182
0.803
C






 T = 0.465
 T = 0.520


rs536355
1
74044111
G/A
G =     
G =     






A =     
A =     


rs553044
1
74044508
G/T
G = 0.471
G = 0.490
0.594
1.079
T






 T = 0.528
 T = 0.509


rs581353
1
74045321
A/G
A = 0.632
A = 0.647
0.666
1.064
G






G = 0.367
G = 0.352


rs496008
1
74045741
T/A
 T = 0.824
 T = 0.822
0.947
0.988
T






A = 0.175
A = 0.177


rs7366854
1
74046600
C/T

  C = 0.847 


  C = 0.854 

0.788
1.057
T






 T = 0.152
 T = 0.145


rs560808
1
74048168
C/A

  C = 0.593 


  C = 0.614 

0.530
1.093
A






A = 0.406
A = 0.385


rs485929
1
74048220
A/G
A = 0.692
A = 0.715
0.481
1.113
G






G = 0.307
G = 0.284


rs477134
1
74048849
T/G
 T = 0.587
 T = 0.612
0.464
1.110
G






G = 0.412
G = 0.387


rs505634
1
74049586
G/A
G = 0.939
G = 0.948
0.664
1.177
A






A = 0.060
A = 0.051


rs542136
1
74049795
G/A
G =     
G =     






A =     
A =     


rs545680
1
74050175
T/C
 T = 0.707
 T = 0.714
0.826
1.035
C







  C = 0.292 


  C = 0.285 



rs522042
1
74051017
G/A
G =     
G =     






A =     
A =     


rs575961
1
74051145
C/T

  C = 0.979 


  C = 0.963 

0.261
0.547
C






 T = 0.020
 T = 0.036


rs792321
1
74051903
A/G
A = 0.667
A = 0.672
0.877
1.025
G






G = 0.332
G = 0.327


rs524306
1
74052188
T/A
 T = 0.517
 T = 0.478
0.260
0.854
T






A = 0.482
A = 0.521


rs495027
1
74052194
A/C
A = 0.562
A = 0.500
0.082
0.778
A







  C = 0.437 


  C = 0.499 



rs525358
1
74053239
A/G
A = 0.127
A = 0.132
0.833
1.050
G






G = 0.872
G = 0.867


rs471496
1
74053841
G/A
G = 0.097
G = 0.096
0.984
0.994
G






A = 0.902
A = 0.903


rs504866
1
74055175
C/T

  C = 0.604 


  C = 0.506 


0.032

0.672
C






 T = 0.395
 T = 0.493


rs532396
1
74056827
T/C
 T = 0.960
 T = 0.963
0.879
1.070
C







  C = 0.039 


  C = 0.036 



rs533371
1
74056947
G/A
G = 0.510
G = 0.510
0.986
1.002
G






A = 0.489
A = 0.489


rs567060
1
74058299
A/G
A = 0.699
A = 0.697
0.966
0.993
A






G = 0.300
G = 0.302


rs4650251
1
74063483
A/G
A = 0.521
A = 0.477
0.213
0.839
A






G = 0.478
G = 0.522


rs3753183
1
74069034
G/T
G = 0.080
G = 0.042

0.031

0.503
G






 T = 0.919
 T = 0.957


rs518769
1
74069847
A/G
A = 0.873
A = 0.870
0.908
0.974
A






G = 0.126
G = 0.129


rs474215
1
74070186
T/C
 T = 0.485
 T = 0.434
0.134
0.813
T







  C = 0.514 


  C = 0.565 



AI
1
74071042
T/C
 T = 0.048
 T = 0.031
0.381
0.635
T







  C = 0.951 


  C = 0.968 



rs500203
1
74071586
T/A
 T = 0.453
 T = 0.423
0.381
0.885
T






A = 0.546
A = 0.576


rs532221
1
74071885
T/A
 T = 0.722
 T = 0.822

˜0.001

1.776
A






A = 0.277
A = 0.177


rs539534
1
74072672
C/T

  C = 0.536 


  C = 0.576 

0.353
1.174
T






 T = 0.463
 T = 0.423


rs531412
1
74072720
G/A
G = 0.740
G = 0.765
0.525
1.141
A






A = 0.259
A = 0.234


rs9970514
1
74073693
C/G

  C = 0.443 


  C = 0.462 

0.576
1.083
G






G = 0.556
G = 0.537


rs568049
1
74074438
C/T

  C = 0.773 


  C = 0.777 

0.914
1.023
T






 T = 0.226
 T = 0.222


rs9727250
1
74077515
T/C
 T = 0.073
 T = 0.059
0.499
0.796
T







  C = 0.926 


  C = 0.940 



rs792323
1
74077529
C/T

  C = 0.820 


  C = 0.824 

0.881
1.029
T






 T = 0.434
 T = 0.406


rs3765651
1
74079297
A/C
A = 0.704
A = 0.704
0.996
0.999
C







  C = 0.295 


  C = 0.295 



rs3765654
1
74079646
C/G

  C = 0.745 


  C = 0.673 


0.031

0.705
C






G = 0.254
G = 0.326


rs481387
1
74080385
C/T

  C = 0.479 


  C = 0.462 

0.622
0.934
C






 T = 0.520
 T = 0.537


rs483259
1
74080564
T/C
 T = 0.636
 T = 0.660
0.478
1.110
C







  C = 0.363 


  C = 0.339 



rs2034453
1
74082116
A/G
A = 0.172
A = 0.150
0.463
0.852
A






G = 0.827
G = 0.849


rs7536997
1
74082887
G/A
G = 0.947
G = 0.955
0.727
1.194
A






A = 0.052
A = 0.044


rs792324
1
74083006
T/C
 T = 0.070
 T = 0.060
0.633
0.850
T






 T = 0.037
 T = 0.044


rs792324
1
74083006
T/C

  C = 0.929 


  C = 0.939 

0.633
0.850
T







  C = 0.962 


  C = 0.955 



rs6660350
1
74083486
C/T

  C = 0.192 


  C = 0.159 

0.259
0.792
C






 T = 0.807
 T = 0.840


rs492302
1
74084074
G/C
G =     
G =     






C =    
C =    


rs10493539
1
74085512
A/G
A = 0.385
A = 0.322
0.124
0.759
A






G = 0.614
G = 0.677


rs496720
1
74088648
G/A
G = 0.960
G = 0.969
0.635
1.285
A






A = 0.039
A = 0.030


rs570631
1
74089790
T/A
 T = 0.384
 T = 0.373
0.769
0.953
T






A = 0.615
A = 0.626


rs4593767
1
74093028
G/A
G = 0.300
G = 0.260
0.236
0.821
G






A = 0.699
A = 0.739


rs576802
1
74094231
T/A
 T = 0.833
 T = 0.865
0.209
1.282
A






A = 0.166
A = 0.134


rs518574
1
74100973
A/G
A = 0.966
A = 0.968
0.880
1.069
G






G = 0.033
G = 0.031


rs524252
1
74102532
C/T

  C = 0.839 


  C = 0.840 

0.966
1.008
T






 T = 0.160
 T = 0.159


rs792327
1
74103456
G/A
G = 0.963
G = 0.978
0.477
1.720
A






A = 0.037
A = 0.022


rs792328
1
74103887
C/A
C =    
C =    






A =     
A =     


rs2039407
1
74104305
A/T
A = 0.945
A = 0.942
0.872
0.937
A






 T = 0.054
 T = 0.057


rs503770
1
74105381
T/C
 T = 0.824
 T = 0.825
0.946
1.013
C







  C = 0.175 


  C = 0.174 



rs503904
1
74105440
A/G
A = 0.814
A = 0.809
0.870
0.970
A






G = 0.185
G = 0.190


rs514012
1
74107016
A/G
A = 0.821
A = 0.801
0.473
0.880
A






G = 0.178
G = 0.198


rs1412827
1
74107532
A/G
A = 0.365
A = 0.387
0.544
1.101
G






G = 0.634
G = 0.612


rs473834
1
74112969
G/A
G = 0.178
G = 0.185
0.803
1.046
A






A = 0.821
A = 0.814


rs480267
1
74113659
A/G
A = 0.332
A = 0.364
0.439
1.153
G






G = 0.667
G = 0.635


rs572180
1
74119134
G/T
G = 0.845
G = 0.850
0.816
1.047
T






 T = 0.154
 T = 0.149


rs1333029
1
74119257
G/A
G = 0.961
G = 0.977
0.294
1.732
A






A = 0.038
A = 0.022


rs485414
1
74119377
A/G
A = 0.897
A = 0.923
0.233
1.376
G






G = 0.102
G = 0.076


rs1412823
1
74119701
A/T
A = 0.761
A = 0.761
0.995
0.999
T






 T = 0.238
 T = 0.238


rs2027013
1
74120888
T/G
 T = 0.093
T =   






G = 0.907
G =     


rs548881
1
74128480
C/T

  C = 0.920 


  C = 0.898 

0.361
0.768
C






 T = 0.079
 T = 0.101


rs792329
1
74130625
C/T

  C = 0.754 


  C = 0.756 

0.951
1.010
T






 T = 0.246
 T = 0.244









Allelotyping results were considered significant with a calculated p-value of less than or equal to 0.05 for allelotype results. These values are indicated in bold. The assay failed for those SNPs in which the allele frequency is blank. The combined allelotyping p-values for males and females were plotted in FIG. 5A and separately for females and males in FIGS. 5B and 5C, respectively. The position of each SNP on the chromosome is presented on the x-axis. The y-axis gives the negative logarithm (base 10) of the p-value comparing the estimated allele frequency in the case group to that of the control group. The minor allele frequency of the control group for each SNP designated by an X or other symbol on the graphs in FIGS. 5A, 5B and 5C can be determined by consulting Table 34 or 35. By proceeding down the Table from top to bottom and across the graphs from left to right the allele frequency associated with each symbol shown can be determined.


To aid the interpretation, multiple lines have been added to the graph. The broken horizontal lines are drawn at two common significance levels, 0.05 and 0.01. The vertical broken lines are drawn every 20 kb to assist in the interpretation of distances between SNPs. Two other lines are drawn to expose linear trends in the association of SNPs to the disease. The light gray line (or generally bottom-most curve) is a nonlinear smoother through the data points on the graph using a local polynomial regression method (W. S. Cleveland, E. Grosse and W. M. Shyu (1992) Local regression models. Chapter 8 of Statistical Models in S eds J. M. Chambers and T. J. Hastie, Wadsworth & Brooks/Cole.). The black line (or generally top-most curve, e.g., see peak in left-most graph just to the left of position 92150000) provides a local test for excess statistical significance to identify regions of association. This was created by use of a 10 kb sliding window with 1 kb step sizes. Within each window, a chi-square goodness of fit test was applied to compare the proportion of SNPs that were significant at a test wise level of 0.01, to the proportion that would be expected by chance alone (0.05 for the methods used here). Resulting p-values that were less than 10−8 were truncated at that value.


Finally, the exons and introns of the genes in the covered region are plotted below each graph at the appropriate chromosomal positions. The gene boundary is indicated by the broken horizontal line. The exon positions are shown as thick, unbroken bars. An arrow is place at the 3′ end of each gene to show the direction of transcription.


Example 10
REPS2 Proximal SNPs

It has been discovered that a polymorphic variation (rs1904528) in the untranslated region of a gene encoding presynaptic cytomatrix protein (REPS2) is associated with the occurrence of melanoma. See Table 3. Subsequently, one-hundred-three allelic variants located within or nearby the target genes were identified and subsequently allelotyped in melanoma case and control sample sets as described in Examples 1 and 2. Several SNPs, designated by SNP_ID “AJ”, “AK”, and “AL” in column 1 of Table 36, are novel and were identified through SNP discovery methods using dye terminator sequencing. Methods of diedeoxynucleotide chain termination chemistry derived sequencing are described by Sanger et al. (Proc. Nat. Acad. Sci. USA 74, 5463-5467, 1997). Briefly, terminators are labeled with fluorescent dyes for automated detection. Fluorescein is measured by an argon ion laser in the sequencing instrument, which transfers light energy to a rhodamine acceptor dye. Each acceptor dye emits light at a characteristic wavelength for detection, identifying the nucleotide that terminated extension of the DNA fragment. Novel SNPs are identified when sequence from multiple samples are compared on one or more strands. The forward and reverse primers used to identify AJ are GCTTTAGGATAGGGAGGAATGG and CCCAGCGCCAGGTAACTAAC, respectively. The forward and reverse primers used to identify AL are CCACCCTTCCTCTGACTGTT and ACAACCCCATCAAAAAGTGG, respectively. The polymorphic variants are set forth in Table 36. The chromosome position provided in column four of Table 36 is based on Genome “Build 34” of NCBI's GenBank and dbSNP build “119”.

TABLE 36PositionAlleleChromo-in SEQChromosomeAllelePresent inIUPACdbSNP rs#someID NO: 7PositionVariantsSEQ ID NO: 7Coders1904525X22316315373T/CCYrs5924557X79016315940C/TCYrs5924616X83416315984A/TTWrs5924617X1,16716316317G/AGRrs7883612X1,37216316522G/AGRrs5924618X2,44816317598C/TCYrs5924619X5,29116320441T/CTYAJX10,53716325687G/TTKrs5901596X13,49516328645—/TGGGNrs7878645X13,86116329011T/CCYrs5924564X14,29916329449C/GCSrs5924535X23,96216339112A/GGRAKX25,21616340366A/CAMrs6629197X33,30416348454C/TTYrs7060904X40,83116355981G/TTKrs4828523X41,42416356574A/GGRrs5924568X41,89816357048C/GGSrs5924569X41,98416357134A/GARrs1904528X50,01316365163C/TGRrs5924570X50,63216365782C/GGSrs1904527X54,49116369641A/GCYrs2052763X55,42216370572G/ACYrs1549529X62,41616377566A/GARrs5924573X64,17216379322A/GARrs5901600X69,27116384421—/TTNrs2288304X70,24416385394T/CTYrs4240072X71,68716386837A/GTYrs3055191X73,57116388721—/GNrs6632943X78,15016393300T/CCYrs5924580X80,03116395181G/AARrs4828544X82,90816398058T/CCYrs1807412X85,21816400368A/GCYrs2010456X85,26916400419G/AGRrs5969758X87,41616402566G/AGRrs1965051X89,28516404435A/GTYre9887218X89,64416404794T/CCYrs1426800X92,64716407797G/ATYrs5969759X102,29416417444A/GGRrs6527766X103,07116418221T/CCYrs1365529X106,61416421764T/CCYrs5924582X107,23116422381G/AARrs2114458X109,07616424226G/AARrs5924583X109,52016424670T/CCYrs5924545X111,21016426360G/AARrs5969761X113,90516429055C/TTYrs6629201X114,43916429589C/TCYrs6629202X114,54416429694C/TTYrs5924584X114,88616430036A/GGRrs5924585X115,26116430411A/GGRrs933946X116,96416432114T/CGRALX119,36816434518C/TCYrs5969762X122,31516437465A/GGRrs713355X126,76816441918T/CGRrs1365530X127,29116442441G/CCSrs1961490X127,36516442515A/TAWrs6527769X128,86616444016C/GCSrs2006847X133,17116448321A/GGRrs2382813X136,37516451525A/GGRrs2033120X136,53116451681C/GCSrs2162394X136,89516452045A/GTYrs1346246X138,42816453578C/AGKrs1365527X139,05216454202G/CGSrs1426801X139,25516454405T/CGRrs5924546X140,61116455761G/CCSrs5924547X142,14416457294G/CCSrs1365528X144,73016459880G/ACYrs4828524X147,46416462614A/GARrs5924588X148,43516463585C/TTYrs5969764X154,00516469155C/TTYrs5924589X158,06816473218T/GTKrs5924590X160,28616475436G/TTKrs5924591X163,77816478928G/TTKrs5924592X165,69716480847A/GARrs5924548X165,74116480891G/AGRrs5924593X165,78516480935C/GGSrs4828547X167,51116482661A/GGRrs714070X168,37816483528A/GCYrs5969768X175,46516490615T/CCYrs4828526X176,48516491635G/AGRrs5969738X177,61316492763G/AARrs5924595X178,20816493358G/AARrs5924549X178,27516493425A/GGRrs4828528X179,55416494704T/CCYrs5969769X180,38216495532G/AARrs4007744X182,50716497657C/TTYrs2891073X184,21416499364T/CCYrs5924599X185,90016501050C/GGSrs1549531X191,01116506161G/AARrs5924601X192,97216508122A/GGRrs5901609X195,13516510285C/—CNrs5969739X196,26616511416C/TCYrs5924602X197,39916512549T/GTKrs1426798X198,61016513760A/CAMrs7891150X198,85116514001C/TTYrs4828550X199,05216514202G/AARrs5924603X200,63916515789T/CTYrs1807947X209,33616524486A/GTYrs954253X209,37916524529A/GARrs2382814X210,01116525161T/GTKrs1896178X210,38016525530A/GCYrs1896177X210,40016525550A/GTYrs5924605X210,55416525704C/GGSrs5924606X213,07116528221A/GAR


Assay for Verifying and Allelotyping SNPs


The methods used to verify and allelotype the one-hundred-three proximal SNPs of Table 36 are the same methods described in Examples 1 and 2 herein. The primers and probes used in these assays are provided in Table 37 and Table 38, respectively.

TABLE 37dbSNP rs #Forward PCR primerReverse PCR primerrs1904525CAAGTTACTGTCTCAGCCTGCAAGTTACTGTCTCAGCCTGrs5924557ACACTCAGGCTACTTCTTACACACTCAGGCTACTTCTTACrs5924616AGCACACGTAATTTGGGTCCAGCACACGTAATTTGGGTCCrs5924617AAAGAGCAGTTCCCCTACACAAAGAGCAGTTCCCCTACACrs7883612CCTCCATGATATGGTTTGTCCCTCCATGATATGGTTTGTCrs5924618TGATCCTCCCACCTCAGGTTTGATCCTCCCACCTCAGGTTrs5924619AGATGGCTGTTGACTGAGAGAGATGGCTGTTGACTGAGAGAJCAGAGTGTTCAGGGTGGTATCAGAGTGTTCAGGGTGGTATrs5901596AAAGTCAGCAACCAGCAGACAAAGTCAGCAACCAGCAGACrs7878645TTGCAGTGAGCCAAGATCACTTGCAGTGAGCCAAGATCACrs5924564GATGAACCTACATTGACACCGATGAACCTACATTGACACCrs5924535AGGTTGCTATGGGAATGGAGAGGTTGCTATGGGAATGGAGAKGGGACAGTTTTCTTGATTTCCGGGACAGTTTTCTTGATTTCCrs6629197ACTTCCCAGACGGGATGGCTACTTCCCAGACGGGATGGCTrs7060904CCAATTGGTAGACAACCAAGCCAATTGGTAGACAACCAAGrs4828523GAGTCTCCTGCTCTTGAAAGGAGTCTCCTGCTCTTGAAAGrs5924568GATGCGATCACTCACGTCTGGATGCGATCACTCACGTCTGrs5924569ACATAGGAAGACCCCCATCTACATAGGAAGACCCCCATCTrs1904528GCTATCTCTTTCACATTGCTCGCTATCTCTTTCACATTGCTCrs5924570TCTAATATGTGTCGGCTCTGTCTAATATGTGTCGGCTCTGrs1904527TCCATCCAGATCATTGCATGTCCATCCAGATCATTGCATGrs2052763TGTTATGAAATGGAGCCACGTGTTATGAAATGGAGCCACGrs1549529AGGATAGTAATGGCCTGGTGAGGATAGTAATGGCCTGGTGrs5924573GGGAGTAGCTTACATCTTGGGGGAGTAGCTTACATCTTGGrs5901600CATACCAAGGATTACAATGTCCATACCAAGGATTACAATGTCrs2288304GGGAGTGGATTCTAATGTTCGGGAGTGGATTCTAATGTTCrs4240072AAGCTAGCCCCCTGATAAAGAAGCTAGCCCCCTGATAAAGrs3055191CCCTCCTCTCTTCTCTACTGCCCTCCTCTCTTCTCTACTGrs6632943TTGACAAAGGAACACTGAGGTTGACAAAGGAACACTGAGGrs5924580CCGCAAACTATTGCAAGGACCCGCAAACTATTGCAAGGACrs4828544TATGAGGCATCAAGAGTAGCTATGAGGCATCAAGAGTAGCrs1807412AGTTCGAAACTAACCTGGCCAGTTCGAAACTAACCTGGCCrs2010456GTGGCTCACTTACGCCTATAGTGGCTCACTTACGCCTATArs5969758AGCTATACCTTGATGCCTGCAGCTATACCTTGATGCCTGCrs1965051TGGTTAAACAGCAGTGTCTGTGGTTAAACAGCAGTGTCTGrs9887218CCATGTTTCCAAAGACAGGGCCATGTTTCCAAAGACAGGGrs1426800GTGGGATTCACTTCCTAAGGGTGGGATTCACTTCCTAAGGrs5969759ACAACTGGCACCATTTTAGCACAACTGGCACCATTTTAGCrs6527766ATTCCACCCGTCAGCATAAGATTCCACCCGTCAGCATAAGrs1365529TTTGCATGCACCCTAAAGAGTTTGCATGCACCCTAAAGAGrs5924582ATGATCGTACCACTGAACCCATGATCGTACCACTGAACCCrs2114458TAATGTGGAGAACTCACAGGTAATGTGGAGAACTCACAGGrs5924583GGAGTTCAAGACCAACCTAGGGAGTTCAAGACCAACCTAGrs5924545TACTTGAGTGGCAAGTCAGGTACTTGAGTGGCAAGTCAGGrs5969761CGTGTTTGTGTTTCAGTATTCGTGTTTGTGTTTCAGTATTrs6629201CACTCACTCTCTGACTTACCCACTCACTCTCTGACTTACCrs6629202CTCTGCCTTTTCTATGTTTCCTCTGCCTTTTCTATGTTTCrs5924584ACTATTGTGTCCCAACTGCCACTATTGTGTCCCAACTGCCrs5924585GTATCCATCCCCTCAAGCATGTATCCATCCCCTCAAGCATrs933946CCTTTAGTTCCTACCTTTGCCCTTTAGTTCCTACCTTTGCALAATACCTGCAGGCAGGTAAGAATACCTGCAGGCAGGTAAGrs5969762TTGGGATTGGAGACACATGGTTGGGATTGGAGACACATGGrs713355ATGTGGTCTATCAGAGCTAGATGTGGTCTATCAGAGCTAGrs1365530CCTCTTCTCAAAACAGTTCTGCCTCTTCTCAAAACAGTTCTGrs1961490TAAAAACCCTGAGGGAGTTCTAAAAACCCTGAGGGAGTTCrs6527769TCTGAACAAAGCTGCAGCTGTCTGAACAAAGCTGCAGCTGrs2006847CACTGGAATCTGATCGTAGCCACTGGAATCTGATCGTAGCrs2382813ACCTAATTCAGCAGAGCACCACCTAATTCAGCAGAGCACCrs2033120TTCTAGGACAGCAAAGGAGGTTCTAGGACAGCAAAGGAGGrs2162394TTTTTGCTGGTAGGGATGGGTTTTTGCTGGTAGGGATGGGrs1346246GTTTATATGCAGCACCTGGGGTTTATATGCAGCACCTGGGrs1365527ATGGTCTGTTTGAGCCTATCATGGTCTGTTTGAGCCTATCrs1426801TGGTAGAACTGCAGTGTAAGTGGTAGAACTGCAGTGTAAGrs5924546ACCTCTGAAAACCTTCTTCGACCTCTGAAAACCTTCTTCGrs5924547CACAGGCATCTGAGGAAATGCACAGGCATCTGAGGAAATGrs1365528CCTTTCTCACCCTTGCAAACCCTTTCTCACCCTTGCAAACrs4828524CCTGGAATGCAGCTGAAATGCCTGGAATGCAGCTGAAATGrs5924588TTAAAGTGCACAATTCAGTGTTAAAGTGCACAATTCAGTGrs5969764GGCAACCACTAACCTATTTTGGGCAACCACTAACCTATTTTGrs5924589ATTTTTGCCATCATCTTCCCATTTTTGCCATCATCTTCCCrs5924590GCTGACTAGACAGATCTGAGGCTGACTAGACAGATCTGAGrs5924591GCACAACTTTCTTCCCAATAGGCACAACTTTCTTCCCAATAGrs5924592TAGCCAGGATGGTCTCGATCTAGCCAGGATGGTCTCGATCrs5924548CAGGTATGTGTCCGTTGTTCCAGGTATGTGTCCGTTGTTCrs5924593ATTACAAGCGTGAGCCATCGATTACAAGCGTGAGCCATCGrs4828547AAGATTCTTCAGATGTGGCCAAGATTCTTCAGATGTGGCCrs714070GACAGCCAAGAGTGAACTAGGACAGCCAAGAGTGAACTAGrs5969768GATATTTGAAAGCTGACAACGATATTTGAAAGCTGACAACrs4828526CAGCAAGGACCTATAGTCCCCAGCAAGGACCTATAGTCCCrs5969738GTCAATTTCGTTGATCTTTGCGTCAATTTCGTTGATCTTTGCrs5924595GACACTTATTGATAAAGAGCGACACTTATTGATAAAGAGCrs5924549GAAGTACTCAGCATTTTGCTCGAAGTACTCAGCATTTTGCTCrs4828528TCCTATGACCCTGCCACATCTCCTATGACCCTGCCACATCrs5969769TCTACGCTTAAACCCATCGCTCTACGCTTAAACCCATCGCrs4007744CCCAAGTAGCTGGGACTACACCCAAGTAGCTGGGACTACArs2891073GACAGCAGAGAAGAGAAATGGACAGCAGAGAAGAGAAATGrs5924599CTCTGAATCCTTGTGTCAGGCTCTGAATCCTTGTGTCAGGrs1549531TTAGGAAACACTGCCTGGTCTTAGGAAACACTGCCTGGTCrs5924601ACACAGCACCTACTACAGTGACACAGCACCTACTACAGTGrs5901609TGAATGAGTAACACTCAGGCTGAATGAGTAACACTCAGGCrs5969739CCCTTCTGATAACTCCTTCGCCCTTCTGATAACTCCTTCGrs5924602ACTTCAAACTGCACAGTCACACTTCAAACTGCACAGTCACrs1426798ATTCACAGAGGCTTAGCAGGATTCACAGAGGCTTAGCAGGrs7891150GGCTCTTCTTTCAGAGAGAGGGCTCITCTTTCAGAGAGAGrs4828550ACTCCAGCCTGGGCAACAAAACTCCAGCCTGGGCAACAAArs5924603GTTGCTAACATCTCCCACAGGTTGCTAACATCTCCCACAGrs1807947AACCATAGGGTGAGTTTGGGAACCATAGGGTGAGTTTGGGrs954253ATGTTCGTCTCACAGTGGACATGTTCGTCTCACAGTGGACrs2382814TCAGCAAGTCCTACCAGTTCTCAGCAAGTCCTACCAGTTCrs1896178AGGTTCACGCCATTCTCCTGAGGTTCACGCCATTCTCCTGrs1896177AGGTTCACGCCATTCTCCTGAGGTTCACGCCATTCTCCTGrs5924605AGAGTGAGACTCTGTCTCAGAGAGTGAGACTCTGTCTCAGrs5924606GAGCCCAAUAACTCCAGTGGAGCCCAATTAACTCCAGTG












TABLE 38













Term












dbSNP rs #
Extend Primer
Mix
















rs1904525
TCCCAGCACTTTGGGAG
ACT








rs5924557
TTCTATCCCCTTACTTCC
ACG







rs5924616
GAAGACAGCAATTTCCT
CGT







rs5924617
TAAGACGTGCCTATGCT
ACG







rs7883612
TTTGTCTGTGTCCCCAC
ACG







rs5924618
CTAGGACTACAGGTGCA
ACG







rs5924619
AAGGACACGACAGAAAG
ACT







AJ
CCTCGTCAGACATGGCTCA
CGT







rs5901596
GGTGGTGGTGTGGTGGTG
ACT







rs7878645
GAGCCAAGATCACGCCACC
ACT







rs5924564
CTCATTATTCCCCAAAGT
ACT







rs5924535
GGGAATGGAGATAAATTCAA
ACT







AK
TTCTTGATTTCCAACTGATT
ACT







rs6629197
ACGGGATGGCTGCCGGG
ACG







rs7060904
GACAACCAAGATGAAAAAC
CGT







rs4828523
AGATTAGCATATTGATAGATTGC
ACT







rs5924568
AGGCAGGAGGATCACTT
ACT







rs5924569
TAGCTGGGCATGTTGGT
ACT







rs1904528
TCACATTGCTCTGCACTTTTG
ACG







rs5924570
CACGAGTGTGTTTTTATCT
ACT







rs1904527
ACTCCTTTTTACTGCTGGGTA
ACT







rs2052763
GAGTGAGTCCCTGTGCA
ACG







rs1549529
TTTGGGAGGCCAAGGTA
ACT







rs5924573
AGCAAAATCTTGGCCGGGC
ACT







rs5901600
GGATTACAATGTCTTTTTTTTTT
ACG







rs2288304
TGAAAAGAGAACAGATTAGTC
ACT







rs4240072
GATGACTGCCACCTGCT
ACT







rs3055191
CACTGGTTTTTTGTTTGTTT
ACT







rs6632943
GGGGGATTGGGTTGGAC
ACT







rs5924580
GTTCTCACTCATAGGTGGGAA
ACG







rs4828544
AAGAGTAGCCAAACTCATA
ACT







rs1807412
CAACATGGTAAAACCCC
ACT







rs2010456
AGGCGGAGGCGGGTGGA
ACG







rs5969758
ATGCCTGCAGGGTACAC
ACG







rs1965051
CAGCAGTGTCTGAGGATTCAA
ACT







rs9887218
CTTGAAGCGTTACCGCCTG
ACT







rs1426800
CTAAGGAAAGCTCATGAC
ACG







rs5969759
TCTTTCCCTTGATCTCAGGC
ACT







rs6527766
CCCAGCACTGCACAAGCTG
ACT







rs1365529
TTTTGTTGGCTGAGCAGAGAT
ACT







rs5924582
GAACCCCAGCCTGGGTGA
ACG







rs2114458
CAGCCCTGACAGAAAGAGC
ACG







rs5924583
GCCAACATGGTGAAACCCC
ACT







rs5924545
GGACCACTGAACCTCTCATT
ACG







rs596976
TTCAGTATTTAACCAAAAAGTTTAAA
ACG







rs6629201
CTTCCAGTCCTGCAAGCTC
ACG







rs6629202
CTTTCTTTCTTCCTTTCTTCCTT
ACG







rs5924584
TATTCAGTACAGGAACATGCC
ACT







rs5924585
CCTTTGAGTTACAAACAATCCA
ACT







rs933946
GATGTTTTTAACAGACTACAGGA
ACT







AL
AGGTAAGGCCTCACCATCAA
ACG







rs5969762
AAGGGAATTAATAGCTGGCAAA
ACT







rs713355
GTTATATTTAACAAGCACAGAAAA
ACT







rs1365530
ATTAATGCCCAACTGATAAAAATA
ACT







rs1961490
AATGTGATTTGGGGCCCTCA
CGT







rs6527769
CTGCAGCTGATTTAGGTCTCA
ACT







rs2006847
GGAAATGACAGGAAGCTGAAC
ACT







rs2382813
GCAGAGCACCATGCACAATG
ACT







rs2033120
GCAGGTAGGAGCAGGTAATG
ACT







rs2162394
GGGGTGAGCCCTGCTGGT
ACT







rs1346246
GGCTAGTACAGAACACATCAA
CGT







rs1365527
GAGCCTATCTTAGTGCCCCT
ACT







rs1426801
GATAGGATAACGAGGATACCAT
ACT







rs5924546
TTAGAGTGGCATTTATGAAGTTA
ACT







rs5924547
CTGAGGAAATGACAGACAGC
ACT







rs1365528
CTCTCCACCCCGACCCC
ACG







rs4828524
GAATGCAGCTGAAATGACAGAA
ACT







rs5924588
TUCAGAATATTTTCATCCCCC
ACG







rs5969764
CTGCAGATTTTCCTTTTCTGAA
ACG







rs5924589
CATCATCTTCCCTCCCCCA
ACT







rs5924590
CTGAGAAAGCAGGGAGTAAC
CGT







rs5924591
TCTACATTTTTACAGCGTAGCA
CGT







rs5924592
GTCTCGATCTCCTGACCTC
ACT







rs5924548
GCCTGGCGCGATGGCTCA
ACG







rs5924593
TATGCCAACTCTTACAATGAGA
ACT







rs4828547
AGATGTGGCCAGATTATTTTAG
ACT







rs714070
GTATGCTTGGTGGCTTATTGA
ACT







rs5969768
ACAGGTTAAGTATCCAAAATCC
ACT







rs4828526
GGACCTATAGTCCCAGCTAC
ACG







rs5969738
CAACTTTTCAACTATTTTCTCTG
ACG







rs5924595
TGATAAAGAGCAAAATGCTGAG
ACG







rs5924549
TCCTAGTAACTAATTCAAAAGAAA
ACT







rs4828528
GGGAACTCAGAGGCTGGCA
ACT







rs5969769
GCGCCATCAAAACAGAACAC
ACG







rs4007744
GCTGGGACTACAGGCGCC
ACG







rs2891073
GAAATGAGGATTCCATTGCTCA
ACT







rs5924599
ATCCTTGTGTCAGGATTCTAC
ACT







rs1549531
ACTGCCTGGTCTATCTCCC
ACG







rs5924601
ACTACAGTGGGCCTCCCC
ACT







rs5901609
ACACTCAGGCCTGAGAATAC
ACG







rs5969739
TAACTCCTTCGACCACAGTC
ACG







rs5924602
AAACTGCACAGTCACCAGAAA
ACT







rs1426798
CTTAGCAGGTGGATGGGGT
ACT







rs7891150
CTTTCAGAGAGAGTGGGCC
ACG







rs4828550
GGGCAACAAAGCGAGACTC
ACG







rs5924603
CACAGAACCTGCTGCTTTTC
ACT







rs1807947
GGTCCACTGTGAGACGAAC
ACT







rs954253
GGACTCCCAAACTCACCCT
ACT







rs2382814
CTACCAGTTCTAGATTCAAGATT
ACT







rs1896178
AGTAGCTGGGACTACAGGC
ACT







rs1896177
CTCCTGCCTCAGCCTCCC
ACT







rs5924605
AACCCTCAATGCCATCCCC
ACT







rs5924606
TAAGCTCAAATTGCAGAAGCC
ACT











Genetic Analysis


Allelotyping results are shown for female (F) and male (M) cases and controls in Table 39 and Table 40, respectively. Allele frequency is noted in the fourth and fifth columns for melanoma pools and control pools, respectively.

TABLE 39FemalesMelanomaChromo-ChromosomeFemaleFemaleFOddsAssociateddbSNP rs#somePositionAllelesCase AFControl AFp-ValueRatioAllelers1904525X16315373T/C T = 0.523 T = 0.5750.1291.233C  C = 0.476   C = 0.424 rs5924557X16315940C/T  C = 0.654   C = 0.598 0.0870.788C T = 0.345 T = 0.401rs5924616X16315984A/TA = 0.562A = 0.5050.0950.797A T = 0.437 T = 0.494rs5924617X16316317G/AG = 0.730G = 0.7080.5010.900GA = 0.269A = 0.291rs7883612X16316522G/AG = 0.515G = 0.5200.8971.017AA = 0.484A = 0.479rs5924618X16317598C/T  C = 0.339   C = 0.274 0.0610.734C T = 0.660 T = 0.725rs5924619X16320441T/C T = 0.471 T = 0.3550.0010.618T  C = 0.528   C = 0.644 AJX16325687G/TG = 0.059G = 0.0300.1230.505G T = 0.940 T = 0.969rs5901596X16328645—/TGG— = 0.351  — = 0.350  0.9790.996TGG = 0.648   TGG = 0.649   rs7878645X16329011T/C T = 0.400 T = 0.4810.0541.395C  C = 0.599   C = 0.518 rs5924564X16329449C/G  C = 0.363   C = 0.260 0.0020.619CG = 0.636G = 0.739rs5924535X16339112A/GA = 0.687A = 0.6450.2070.826AG = 0.312G = 0.354AKX16340366A/CA = 0.744A = 0.6840.0500.743A  C = 0.255   C = 0.315 rs6629197X16348454C/T  C = 0.739   C = 0.758 0.5251.102T T = 0.260 T = 0.241rs7060904X16355981G/TG = 0.478G = 0.5230.1891.194T T = 0.521 T = 0.476rs4828523X16356574A/GA = 0.858A = 0.9060.0401.592GG = 0.141G = 0.093rs5924568X16357048C/G  C = 0.825   C = 0.873 0.0571.453GG = 0.174G = 0.126rs5924569X16357134A/GA = 0.377A = 0.266˜0.0010.599AG = 0.622G = 0.733rs1904528X16365163C/T  C = 0.254   C = 0.200 0.0540.736C T = 0.745 T = 0.799rs5924570X16365782C/G  C = 0.390   C = 0.390 0.9941.001G = 0.609G = 0.609rs1904527X16369641A/GA = 0.543A = 0.4780.0820.771AG = 0.456G = 0.521rs2052763X16370572G/AG = 0.298G = 0.2190.0060.661GA = 0.701A = 0.780rs1549529X16377566A/GA = 0.437A = 0.311˜0.0010.580AG = 0.562G = 0.688rs5924573X16379322A/GA = 0.521A = 0.4130.0080.646AG = 0.478G = 0.586rs5901600X16384421—/T— = 0.849  — = 0.873  0.3291.222T— = 0.855  — = 0.885  rs5901600X16384421—/T T = 0.150 T = 0.1260.3291.222T T = 0.144 T = 0.114rs2288304X16385394T/C T = 0.729 T = 0.7010.3920.871T  C = 0.270   C = 0.298 rs4240072X16386837A/GA = 0.346A = 0.229˜0.0010.562AG = 0.653G = 0.770rs3055191X16388721—/— = 0.499  — = 0.488  0.7700.959 = 0.500 = 0.511rs6632943X16393300T/C T = 0.461 T = 0.5200.0801.268C  C = 0.538   C = 0.479 rs5924580X16395181G/AG = 0.847G = 0.7940.0490.693GA = 0.152A = 0.205rs4828544X16398058T/C T = 0.783 T = 0.8550.0061.633C  C = 0.216   C = 0.144 rs1807412X16400368A/GA = 0.596A = 0.6200.5061.107GG = 0.403G = 0.379rs2010456X16400419G/AG = 0.022G = 0.0150.6070.699GA = 0.978A = 0.985rs5969758X16402566G/AG = 0.248G = 0.1920.0410.719GA = 0.751A = 0.807rs1965051X16404435A/GA = 0.341A = 0.232˜0.0010.585AG = 0.658G = 0.767rs9887218X16404794T/C T = 0.007 T = 0.0430.0696.157C  C = 0.993   C = 0.957 rs1426800X16407797G/AG = 0.752G = 0.860˜0.0012.027AA = 0.247A = 0.139rs5969759X16417444A/GA = 0.631A = 0.6810.1241.245GG = 0.368G = 0.318rs6527766X16418221T/C T = 0.801 T = 0.8120.7031.069C  C = 0.198   C = 0.187 rs1365529X16421764T/C T = 0.824 T = 0.8900.0081.738C  C = 0.175   C = 0.109 rs5924582X16422381G/AG = 0.800G = 0.8270.3241.199AA = 0.199A = 0.172rs2114458X16424226G/AG = 0.808G = 0.8520.0811.366AA = 0.191A = 0.147rs5924583X16424670T/C T = 0.701 T = 0.7260.4211.132C  C = 0.298   C = 0.273 rs5924545X16426360G/AG = 0.813G = 0.8680.0291.510AA = 0.186A = 0.131rs5969761X16429055C/T  C = 0.760   C = 0.901 ˜0.0012.881T T = 0.239 T = 0.098rs6629201X16429589C/T  C = 0.657   C = 0.625 0.3390.871C T = 0.342 T = 0.374rs6629202X16429694C/T  C = 0.704   C = 0.786 0.0131.544T T = 0.295 T = 0.213rs5924584X16430036A/GA = 0.795A = 0.8730.0011.779GG = 0.204G = 0.126rs5924585X16430411A/GA = 0.870A = 0.8680.9290.981AG = 0.129G = 0.131rs933946X16432114T/C T = 0.835 T = 0.8660.2021.278C  C = 0.164   C = 0.133 ALX16434518C/T  C = 0.820   C = 0.799 0.4220.870C T = 0.179 T = 0.200rs5969762X16437465A/GA = 0.563A = 0.5770.7311.055GG = 0.436G = 0.422rs713355X16441918T/C T = 0.559 T = 0.5590.9951.001  C = 0.440   C = 0.440 rs1365530X16442441G/CG = 0.798G = 0.8700.0071.699C  C = 0.201   C = 0.129 rs1961490X16442515A/TA = 0.768A = 0.7220.1210.783A T = 0.231 T = 0.277rs6527769X16444016C/G  C = 0.388   C = 0.290 0.0040.647CG = 0.611G = 0.709rs2006847X16448321A/GA = 0.555A = 0.5560.9811.003GG = 0.444G = 0.443rs2382813X16451525A/GA = 0.876A = 0.9230.0351.693GG = 0.123G = 0.076rs2033120X16451681C/G  C = 0.790   C = 0.864 0.0081.695GG = 0.209G = 0.135rs2162394X16452045A/GA = 0.310A = 0.2210.0020.632AG = 0.689G = 0.778rs1346246X16453578C/A  C = 0.400   C = 0.303 0.0020.652CA = 0.599A = 0.696rs1365527X16454202G/CG = 0.817G = 0.8700.0401.495C  C = 0.182   C = 0.129 rs1426801X16454405T/C T = 0.432 T = 0.4770.3211.200C  C = 0.567   C = 0.522 rs5924546X16455761G/CG = 0.366G = 0.4190.1061.252C  C = 0.633   C = 0.580 rs5924547X16457294G/CG = 0.562G = 0.5410.5190.917G  C = 0.437   C = 0.458 rs1365528X16459880G/AG = 0.221G = 0.1800.1330.775GA = 0.778A = 0.819rs4828524X16462614A/GA = 0.296A = 0.2050.0010.614AG = 0.703G = 0.794rs5924588X16463585C/T  C = 0.676   C = 0.788 ˜0.0011.784T T = 0.323 T = 0.211rs5969764X16469155C/T  C = 0.784   C = 0.861 0.0031.720T T = 0.215 T = 0.138rs5924589X16473218T/G T = 0.340 T = 0.2580.0120.678TG = 0.659G = 0.741rs5924590X16475436G/TG = 0.809G = 0.8690.0161.572T T = 0.190 T = 0.130rs5924591X16478928G/TG = 0.844G = 0.8870.0641.460T T = 0.155 T = 0.112rs5924592X16480847A/GA = 0.423A = 0.4220.9800.997AG = 0.576G = 0.577rs5924548X16480891G/AG = 0.748G = 0.6980.1040.778GA = 0.251A = 0.301rs5924593X16480935C/G  C = 0.723   C = 0.782 0.0511.380GG = 0.276G = 0.217rs4828547X16482661A/GA = 0.807A = 0.8590.0411.457GG = 0.192G = 0.140rs714070X16483528A/GA = 0.566A = 0.5520.6850.945AG = 0.433G = 0.447rs5969768X16490615T/C T = 0.077 T = 0.0780.9591.014C  C = 0.922   C = 0.921 rs4828526X16491635G/AG = 0.221G = 0.1570.0210.655GA = 0.778A = 0.842rs5969738X16492763G/AG = 0.538G = 0.5660.4311.117AA = 0.461A = 0.433rs5924595X16493358G/AG = 0.772G = 0.8480.0111.655AA = 0.227A = 0.151rs5924549X16493425A/GA = 0.699A = 0.7770.0111.502GG = 0.300G = 0.222rs4828528X16494704T/C T = 0.317 T = 0.3160.9730.995T  C = 0.682   C = 0.683 rs5969769X16495532G/AG = 0.800G = 0.8780.0031.801AA = 0.199A = 0.121rs4007744X16497657C/T  C = 0.532   C = 0.636 0.0061.537T T = 0.467 T = 0.363rs2891073X16499364T/C T = 0.368 T = 0.172˜0.0010.356T  C = 0.631   C = 0.827 rs5924599X16501050C/G  C = 0.531   C = 0.535 0.9201.013GG = 0.468G = 0.464rs1549531X16506161G/AG = 0.719G = 0.819˜0.0011.774AA = 0.280A = 0.180rs5924601X16508122A/GA = 0.148A = 0.1530.8131.047GG = 0.851G = 0.846rs5901609X16510285C/—  C = 0.269   C = 0.175 0.0010.578C— = 0.730  — = 0.824  rs5969739X16511416C/T  C = 0.297   C = 0.223 0.0110.680C T = 0.702 T = 0.776rs5924602X16512549T/G T = 0.617 T = 0.6030.6620.941TG = 0.382G = 0.396rs1426798X16513760A/CA = 0.337A = 0.2570.0080.681A  C = 0.662   C = 0.742 rs7891150X16514001C/T  C = 0.986   C = 0.966 0.2270.409C T = 0.014 T = 0.034rs4828550X16514202G/AG = 0.758G = 0.850˜0.0011.820AA = 0.241A = 0.149rs5924603X16515789T/C T = 0.319 T = 0.2380.0110.665T  C = 0.680   C = 0.761 rs1807947X16524486A/GA = 0.347A = 0.2620.0050.667AG = 0.652G = 0.737rs954253X16524529A/GA = 0.701A = 0.6850.6010.927AG = 0.298G = 0.314rs2382814X16525161T/G T = 0.327 T = 0.224˜0.0010.595TG = 0.672G = 0.775rs1896178X16525530A/GA = 0.553A = 0.5200.3240.873AG = 0.446G = 0.479rs1896177X16525550A/GA = 0.499A = 0.5030.9171.014GG = 0.500G = 0.496rs5924605X16525704C/G  C = 0.805   C = 0.839 0.2241.262GG = 0.194G = 0.160rs5924606X16528221A/GA = 0.443A = 0.3530.0080.688AG = 0.556G = 0.646









TABLE 40










Males























Melanoma



Chromo-
Chromosome

Male
Male
F
Odds
Associated


dbSNP rs#
some
Position
Alleles
Case AF
Control AF
p-Value
Ratio
Allele


















rs1904525
X
16315373
T/C
 T = 0.569
 T = 0.597
0.487
1.122
C







  C = 0.430 


  C = 0.402 



rs5924557
X
16315940
C/T

  C = 0.592 


  C = 0.591 

0.975
0.996
C






 T = 0.407
 T = 0.408


rs5924616
X
16315984
A/T
A = 0.495
A = 0.456
0.292
0.854
A






 T = 0.504
 T = 0.543


rs5924617
X
16316317
G/A
G =     
G =     






A =     
A =     


rs7883612
X
16316522
G/A
G = 0.559
G = 0.529
0.371
0.884
G






A = 0.440
A = 0.470


rs5924618
X
16317598
C/T

  C = 0.386 


  C = 0.369 

0.609
0.930
C






 T = 0.613
 T = 0.630


rs5924619
X
16320441
T/C
 T = 0.437
 T = 0.369

0.047

0.753
T







  C = 0.562 


  C = 0.630 



AJ
X
16325687
G/T
G = 0.038
G = 0.015
0.185
0.385
G






 T = 0.962
 T = 0.985


rs5901596
X
16328645
—/TGG
— = 0.351  
— = 0.340  
0.728
0.950







TGG = 0.648   
TGG = 0.659   


rs7878645
X
16329011
T/C
 T = 0.452
 T = 0.473
0.575
1.086
C







  C = 0.547 


  C = 0.526 



rs5924564
X
16329449
C/G

  C = 0.325 


  C = 0.258 


0.042

0.722
C






G = 0.674
G = 0.741


rs5924535
X
16339112
A/G
A = 0.634
A = 0.633
0.997
1.000
A






G = 0.365
G = 0.366


AK
X
16340366
A/C
A = 0.687
A = 0.705
0.605
1.088
C







  C = 0.312 


  C = 0.294 



rs6629197
X
16348454
C/T

  C = 0.760 


  C = 0.761 

0.969
1.006
T






 T = 0.239
 T = 0.238


rs7060904
X
16355981
G/T
G = 0.479
G = 0.525
0.187
1.200
T






 T = 0.520
 T = 0.474


rs4828523
X
16356574
A/G
A = 0.873
A = 0.903
0.191
1.367
G






G = 0.126
G = 0.096


rs5924568
X
16357048
C/G

  C = 0.847 


  C = 0.885 

0.117
1.390
G






G = 0.152
G = 0.114


rs5924569
X
16357134
A/G
A = 0.324
A = 0.266
0.088
0.756
A






G = 0.675
G = 0.733


rs1904528
X
16365163
C/T

  C = 0.244 


  C = 0.207 

0.219
0.810
C






 T = 0.755
 T = 0.792


rs5924570
X
16365782
C/G

  C = 0.424 


  C = 0.409 

0.665
0.941
C






G = 0.575
G = 0.590


rs1904527
X
16369641
A/G
A = 0.500
A = 0.470
0.468
0.888
A






G = 0.499
G = 0.529


rs2052763
X
16370572
G/A
G = 0.278
G = 0.214

0.034

0.709
G






A = 0.721
A = 0.785


rs1549529
X
16377566
A/G
A = 0.396
A = 0.250

˜0.001

0.510
A






G = 0.603
G = 0.749


rs5924573
X
16379322
A/G
A = 0.479
A = 0.408
0.057
0.751
A






G = 0.520
G = 0.591


rs5901600
X
16384421
—/T
— = 0.831  
— = 0.863  
0.211
1.283
T






— = 0.867  
— = 0.890  


rs5901600
X
16384421
—/T
 T = 0.168
 T = 0.136
0.211
1.283
T






 T = 0.132
 T = 0.109


rs2288304
X
16385394
T/C
 T = 0.676
 T = 0.689
0.726
1.062
C







  C = 0.323 


  C = 0.310 



rs4240072
X
16386837
A/G
A = 0.305
A = 0.227

0.018

0.668
A






G = 0.694
G = 0.772


rs3055191
X
16388721
—/
— = 0.497  
— = 0.479  
0.621
0.933







 = 0.502
 = 0.520


rs6632943
X
16393300
T/C
 T = 0.504
 T = 0.503
0.967
0.994
T







  C = 0.495 


  C = 0.496 



rs5924580
X
16395181
G/A
G = 0.811
G = 0.775
0.264
0.803
G






A = 0.188
A = 0.224


rs4828544
X
16398058
T/C
 T = 0.797
 T = 0.852

0.039

1.464
C







  C = 0.202 


  C = 0.147 



rs1807412
X
16400368
A/G
A = 0.582
A = 0.629
0.202
1.214
G






G = 0.417
G = 0.370


rs2010456
X
16400419
G/A
G = 0.023
G = 0.026
0.799
1.154
A






A = 0.976
A = 0.973


rs5969758
X
16402566
G/A
G = 0.238
G = 0.200
0.193
0.801
G






A = 0.761
A = 0.799


rs1965051
X
16404435
A/G
A = 0.305
A = 0.239

0.036

0.716
A






G = 0.694
G = 0.760


rs9887218
X
16404794
T/C
 T = 0.040
 T = 0.038
0.946
0.959
T







  C = 0.959 


  C = 0.961 



rs1426800
X
16407797
G/A
G = 0.798
G = 0.871

0.005

1.711
A






A = 0.201
A = 0.128


rs5969759
X
16417444
A/G
A = 0.621
A = 0.672
0.165
1.252
G






G = 0.378
G = 0.327


rs6527766
X
16418221
T/C
 T = 0.787
 T = 0.799
0.704
1.073
C







  C = 0.212 


  C = 0.200 



rs1365529
X
16421764
T/C
 T = 0.836
 T = 0.874
0.131
1.359
C







  C = 0.163 


  C = 0.125 



rs5924582
X
16422381
G/A
G = 0.780
G = 0.797
0.542
1.109
A






A = 0.219
A = 0.202


rs2114458
X
16424226
G/A
G = 0.814
G = 0.838
0.381
1.186
A






A = 0.185
A = 0.161


rs5924583
X
16424670
T/C
 T = 0.715
 T = 0.726
0.711
1.059
C







  C = 0.284 


  C = 0.273 



rs5924545
X
16426360
G/A
G = 0.822
G = 0.860
0.141
1.333
A






A = 0.177
A = 0.139


rs5969761
X
16429055
C/T

  C = 0.811 


  C = 0.899 


0.001

2.076
T






 T = 0.188
 T = 0.100


rs6629201
X
16429589
C/T

  C = 0.609 


  C = 0.632 

0.486
1.103
T






 T = 0.390
 T = 0.367


rs6629202
X
16429694
C/T

  C = 0.733 


  C = 0.785 

0.151
1.328
T






 T = 0.266
 T = 0.214


rs5924584
X
16430036
A/G
A = 0.826
A = 0.878

0.039

1.517
G






G = 0.173
G = 0.121


rs5924585
X
16430411
A/G
A = 0.851
A = 0.851
0.997
0.999






G = 0.148
G = 0.148


rs933946
X
16432114
T/C
 T = 0.834
 T = 0.855
0.434
1.173
C







  C = 0.165 


  C = 0.144 



AL
X
16434518
C/T

  C = 0.765 


  C = 0.779 

0.673
1.078
T






 T = 0.234
 T = 0.220


rs5969762
X
16437465
A/G
A = 0.599
A = 0.564
0.308
0.866
A






G = 0.400
G = 0.435


rs713355
X
16441918
T/C
 T = 0.521
 T = 0.542
0.551
1.086
C







  C = 0.478 


  C = 0.457 



rs1365530
X
16442441
G/C
G = 0.816
G = 0.874

0.024

1.577
C







  C = 0.183 


  C = 0.125 



rs1961490
X
16442515
A/T
A = 0.711
A = 0.723
0.701
1.061
T






 T = 0.288
 T = 0.276


rs6527769
X
16444016
C/G

  C = 0.353 


  C = 0.290 

0.073
0.746
C






G = 0.646
G = 0.709


rs2006847
X
16448321
A/G
A = 0.514
A = 0.544
0.410
1.128
G






G = 0.485
G = 0.455


rs2382813
X
16451525
A/G
A = 0.893
A = 0.912
0.379
1.238
G






G = 0.106
G = 0.087


rs2033120
X
16451681
C/G

  C = 0.810 


  C = 0.851 

0.161
1.338
G






G = 0.189
G = 0.148


rs2162394
X
16452045
A/G
A = 0.295
A = 0.238
0.064
0.744
A






G = 0.704
G = 0.761


rs1346246
X
16453578
C/A

  C = 0.383 


  C = 0.312 


0.031

0.729
C






A = 0.616
A = 0.687


rs1365527
X
16454202
G/C
G = 0.830
G = 0.864
0.185
1.303
C







  C = 0.169 


  C = 0.135 



rs1426801
X
16454405
T/C
 T = 0.450
 T = 0.461
0.791
1.043
C







  C = 0.549 


  C = 0.538 



rs5924546
X
16455761
G/C
G = 0.419
G = 0.419
0.994
1.001







  C = 0.580 


  C = 0.580 



rs5924547
X
16457294
G/C
G = 0.523
G = 0.551
0.437
1.115
C







  C = 0.476 


  C = 0.448 



rs1365528
X
16459880
G/A
G = 0.206
G = 0.180
0.381
0.848
G






A = 0.793
A = 0.819


rs4828524
X
16462614
A/G
A = 0.271
A = 0.211

0.049

0.719
A






G = 0.728
G = 0.788


rs5924588
X
16463585
C/T

  C = 0.713 


  C = 0.785 


0.026

1.469
T






 T = 0.286
 T = 0.214


rs5969764
X
16469155
C/T

  C = 0.817 


  C = 0.877 

0.066
1.605
T






 T = 0.182
 T = 0.122


rs5924589
X
16473218
T/G
 T = 0.319
 T = 0.267
0.104
0.778
T






G = 0.680
G = 0.732


rs5924590
X
16475436
G/T
G = 0.831
G = 0.871
0.121
1.369
T






 T = 0.168
 T = 0.128


rs5924591
X
16478928
G/T
G = 0.850
G = 0.885
0.166
1.361
T






 T = 0.149
 T = 0.114


rs5924592
X
16480847
A/G
A = 0.416
A = 0.413
0.938
0.989
A






G = 0.583
G = 0.586


rs5924548
X
16480891
G/A
G = 0.697
G = 0.707
0.774
1.045
A






A = 0.302
A = 0.292


rs5924593
X
16480935
C/G

  C = 0.716 


  C = 0.815 


0.010

1.744
G






G = 0.283
G = 0.184


rs4828547
X
16482661
A/G
A = 0.810
A = 0.847
0.162
1.300
G






G = 0.189
G = 0.152


rs714070
X
16483528
A/G
A = 0.525
A = 0.550
0.490
1.104
G






G = 0.474
G = 0.449


rs5969768
X
16490615
T/C
 T = 0.054
 T = 0.166

˜0.001

3.445
C







  C = 0.945 


  C = 0.833 



rs4828526
X
16491635
G/A
G = 0.211
G = 0.174
0.223
0.791
G






A = 0.788
A = 0.825


rs5969738
X
16492763
G/A
G = 0.492
G = 0.545
0.136
1.237
A






A = 0.507
A = 0.454


rs5924595
X
16493358
G/A
G = 0.782
G = 0.842

0.038

1.487
A






A = 0.217
A = 0.157


rs5924549
X
16493425
A/G
A = 0.727
A = 0.774
0.129
1.287
G






G = 0.272
G = 0.225


rs4828528
X
16494704
T/C
 T = 0.182
 T = 0.179
0.921
0.980
T







  C = 0.817 


  C = 0.820 



rs5969769
X
16495532
G/A
G = 0.823
G = 0.874
0.061
1.494
A






A = 0.176
A = 0.125


rs4007744
X
16497657
C/T

  C = 0.597 


  C = 0.644 

0.252
1.221
T






 T = 0.402
 T = 0.355


rs2891073
X
16499364
T/C
 T = 0.256
 T = 0.099

˜0.001

0.321
T







  C = 0.743 


  C = 0.900 



rs5924599
X
16501050
C/G

  C = 0.479 


  C = 0.554 


0.033

1.347
G






G = 0.520
G = 0.445


rs1549531
X
16506161
G/A
G = 0.748
G = 0.821

0.010

1.547
A






A = 0.251
A = 0.178


rs5924601
X
16508122
A/G
A = 0.132
A = 0.231

˜0.001

1.975
G






G = 0.867
G = 0.768


rs5901609
X
16510285
C/—

  C = 0.253 


  C = 0.190 


0.032

0.689
C






— = 0.746  
— = 0.809  


rs5969739
X
16511416
C/T

  C = 0.278 


  C = 0.246 

0.293
0.846
C






 T = 0.721
 T = 0.753


rs5924602
X
16512549
T/G
 T = 0.666
 T = 0.588

0.020

0.716
T






G = 0.333
G = 0.411


rs1426798
X
16513760
A/C
A = 0.336
A = 0.270

0.040

0.729
A







  C = 0.663 


  C = 0.729 



rs7891150
X
16514001
C/T

  C = 0.979 


  C = 0.970 

0.622
0.674
C






 T = 0.020
 T = 0.029


rs4828550
X
16514202
G/A
G = 0.763
G = 0.848

0.004

1.730
A






A = 0.236
A = 0.151


rs5924603
X
16515789
T/C
 T = 0.314
 T = 0.241

0.019

0.692
T







  C = 0.685 


  C = 0.758 



rs1807947
X
16524486
A/G
A = 0.340
A = 0.260

0.013

0.681
A






G = 0.659
G = 0.739


rs954253
X
16524529
A/G
A = 0.659
A = 0.681
0.514
1.101
G






G = 0.340
G = 0.318


rs2382814
X
16525161
T/G
 T = 0.293
 T = 0.231
0.059
0.726
T






G = 0.706
G = 0.768


rs1896178
X
16525530
A/G
A = 0.541
A = 0.511
0.404
0.887
A






G = 0.458
G = 0.488


rs1896177
X
16525550
A/G
A = 0.498
A = 0.501
0.931
1.012
G






G = 0.501
G = 0.498


rs5924605
X
16525704
C/G

  C = 0.802 


  C = 0.839 

0.214
1.289
G






G = 0.197
G = 0.160


rs5924606
X
16528221
A/G
A = 0.419
A = 0.340

0.025

0.713
A






G = 0.580
G = 0.659









Allelotyping results were considered significant with a calculated p-value of less than or equal to 0.05 for allelotype results. These values are indicated in bold. The assay failed for those SNPs in which the allele frequency is blank. The combined allelotyping p-values for males and females were plotted in FIG. 6A and separately for females and males in FIGS. 6B and 6C, respectively. The position of each SNP on the chromosome is presented on the x-axis. The y-axis gives the negative logarithm (base 10) of the p-value comparing the estimated allele frequency in the case group to that of the control group. The minor allele frequency of the control group for each SNP designated by an X or other symbol on the graphs in FIGS. 6A, 6B and 6C can be determined by consulting Table 39 or 40. By proceeding down the Table from top to bottom and across the graphs from left to right the allele frequency associated with each symbol shown can be determined.


To aid the interpretation, multiple lines have been added to the graph. The broken horizontal lines are drawn at two common significance levels, 0.05 and 0.01. The vertical broken lines are drawn every 20 kb to assist in the interpretation of distances between SNPs. Two other lines are drawn to expose linear trends in the association of SNPs to the disease. The light gray line (or generally bottom-most curve) is a nonlinear smoother through the data points on the graph using a local polynomial regression method (W. S. Cleveland, E. Grosse and W. M. Shyu (1992) Local regression models. Chapter 8 of Statistical Models in S eds J. M. Chambers and T. J. Hastie, Wadsworth & Brooks/Cole.). The black line (or generally top-most curve, e.g., see peak in left-most graph just to the left of position 92150000) provides a local test for excess statistical significance to identify regions of association. This was created by use of a 1 Okb sliding window with 1 kb step sizes. Within each window, a chi-square goodness of fit test was applied to compare the proportion of SNPs that were significant at a test wise level of 0.01, to the proportion that would be expected by chance alone (0.05 for the methods used here). Resulting p-values that were less than 10−8 were truncated at that value.


Finally, the exons and introns of the genes in the covered region are plotted below each graph at the appropriate chromosomal positions. The gene boundary is indicated by the broken horizontal line. The exon positions are shown as thick, unbroken bars. An arrow is place at the 3′ end of each gene to show the direction of transcription.


Example 11
Inhibition of NRP1 Gene Expression by Transfection of Specific siRNAs

RNAi-based gene inhibition was selected as an effective way to inhibit expression of NRP1 in cultured cells. Algorithms useful for designing siRNA molecules specific for the NRP1 targets are disclosed at the http address www.dhramacon.com. siRNAs were selected from this list for use in RNAi experiments following a filtering protocol that involved the removal of any siRNA with complementarity to common motifs or domains present in any target as well as siRNAs complementary to sequences containing SNPs. From this filtered set of siRNAs, four were selected that showed no off-target homology following BLAST analysis against various Genbank nucleotide databases. Table 41 summarize the features of the duplexes that were ordered from Dharmacon Research, Inc., and subsequently used as a cocktail in the assays described herein to inhibit expression of NRP1, NID2 ENDO180, CDK10, FPGT, CARK, PCLO or REPS2, respectively. A non-homologous siRNA reagent was used as a negative control.

TABLE 41NRP1 siRNAs used for cell transfectionsiRNASEQ IDsiRNATargetSequence SpecificityNO:NRP1_425NRP1AGAGAGGTCCTGAATGTTCNRP1_1282NRP1GATTATCCTTGCTCTGGAANRP1_1910NRP1GCACCATACAATCAGAGTTNRP1_2538NRP1GCCAGGCAATGTGTTGAAG


Two melanoma cell lines (M14 and A375) and a breast cancer cell line (MCF7) were selected for RNAi experiments. On day 1, cells were transfected with siRNA cocktails (18.75 nM) using LIPOFECTAMINE 2000 (LF2000™) Reagent. On days 1, 3 and 6, cellular proliferation was measured using the WST-1 assay (Roche, catalog #1644807). Briefly, the WST-1 assay is a colorimetric assay used to determine cellular proliferation by measuring the cleavage of WST-1 by mitochondrial dehydrogenases in living cells. By measuring absorbance, a highly accurate measure of cellular proliferation is obtained. On day 1, WST-1 reagent was added to each well and allowed to incubate for 3-4 hours. Subsequently, absorbance was measured at 450 nm and 620 nm using a Tecan Ultra plate reader. This process was repeated on day's 3 and 6. The extent of proliferation on days 3 and 6 were calculated relative to day 1, based on absorbance readings for each sample on each day. From the triplicate repeats of each time point, means and standard deviations were calculated and the effect of siRNA inhibition of each target on cellular proliferation was assessed and compared to cells transfected with a positive control siRNA (siRAD211175) and a negative control siRNA (siLuciferase GL2). All experiments were performed in duplicate.


The difference in absorbance between these 2 wavelengths is an indication of the metabolic activity in each well that was measured. Metabolic activity is directly proportional to the number of cells in each well. Suppression of target mRNA levels correlated with decreased cell proliferation. The NRP1 siRNA suppressed proliferation of melanoma M14 cells and A375 cell (see FIGS. 7A and 7B respectively).


Example 12
Inhibition of CDK10 Gene Expression by Transfection of Specific siRNAs

RNAi-based gene inhibition was selected as an effective way to inhibit expression of CDK10 in cultured cells. Algorithms useful for designing siRNA molecules specific for the CDK10 targets are disclosed at the http address www.dhramacon.com. siRNAs were selected from this list for use in RNAi experiments following a filtering protocol that involved the removal of any siRNA with complementarity to common motifs or domains present in any target as well as siRNAs complementary to sequences containing SNPs. From this filtered set of siRNAs, four were selected that showed no off-target homology following BLAST analysis against various Genbank nucleotide databases. Table 42 summarizes the features of the duplexes that were ordered from Dharmacon Research, Inc., and subsequently used as a cocktail in the assays described herein to inhibit expression of CDK10. A non-homologous siRNA reagent was used as a negative control.

TABLE 42CDK10 siRNAs used for cell transfectionsiRNAsiRNATargetSequence SpecificitySEQ ID NO:CDK10_59CDK10GATCCGTCTGAAGTGTATTCDK10_149CDK10GAAGCTGAACCGCATTGGACDK10_175CDK10CCTACGGCATTGTGTATCGCDK10_532CDK10ACTTGCTCATGACCGACAA


Two melanoma cell lines (M14 and A375) were selected for RNAi experiments. On day 1, cells were transfected with siRNA cocktails (18.75 nM) using LIPOFECTAMINE 2000 (LF2000™) Reagent. On days 1, 3 & 6, cellular proliferation was measured using the WST-1 assay (Roche, catalog #1 644 807). Briefly, the WST-1 assay is a colorimetric assay used to determine cellular proliferation by measuring the cleavage of WST-1 by mitochondrial dehydrogenases in living cells. By measuring absorbance, a highly accurate measure of cellular proliferation is obtained. On day 1, WST-1 reagent was added to each well and allowed to incubate for 3-4 h. Subsequently, absorbance was measured at 450 nm and 620 nm using a Tecan Ultra plate reader. This process was repeated on day's 3 and 6. The extent of proliferation on days 3 and 6 were calculated relative to day 1, based on absorbance readings for each sample on each day. From the triplicate repeats of each time point, means and standard deviations were calculated and the effect of siRNA inhibition of each target on cellular proliferation was assessed and compared to cells transfected with a positive control siRNA (siRAD211175) and a negative control siRNA (siLuciferase GL2). All experiments were performed in duplicate.


Example 13
Cell Proliferation

The siRNAs from Example 12 were transfected in cell lines grown in 6-well plates. Cells were trypsinized on the following day and distributed into 96-well plates. Wst-1 reagent was added on the indicated days and the absorbance at 650 nm and 450 nm was measured. The difference in absorbance between these 2 wavelengths is an indication of the metabolic activity in each well that was measured. Metabolic activity is directly proportional to the number of cells in each well.


Suppression of target mRNA levels correlated with decreased cell proliferation as seen in A375 melanoma cells (see FIG. 8).


Example 14
Screening Assay to Detect Modulators of NRP1

The following is an exemplary assay for finding modulators of NRP1. Plasma membranes are prepared from cells expressing NRP1 by standard methods. The membranes are immobilized on wheat germ agglutinin-coated scintillation proximity beads (Amersham). Test samples and radiolabeled ligand, which is 125I-semaphorin-3, are added, and bead-bound radioactivity is measured. The ability of a given sample to modulate binding activity is determined by comparison of bead-bound radioactivity in the presence of the compound with values obtained in solvent controls. The test compound optimally produces a result that differs by at least 3 standard deviations from a set of at least 30 replicate control samples.


Example 15
Screening Assay to Detect Modulators of NID2

The following is an exemplary assay for finding modulators of NID2. Plasma membranes as prepared are prepared from cells expressing integrin alpha-3-beta-1 or integrin alpha-6-beta-1 by standard methods. The membranes are immobilized on wheat germ agglutinin-coated scintillation proximity beads (Amersham). Test sample and radiolabeled ligand, which is 125I-NID2, are added, and bead-bound radioactivity is measured. The ability of a given sample to modulate binding activity is determined by comparison of bead-bound radioactivity in the presence of the compound with values obtained in solvent controls. The test compound optimally produces a result that differs by at least 3 standard deviations from a set of at least 30 replicate control samples.


Example 16
Screening Assay to Detect Modulators of ENDO180

The following is an exemplary assay for finding modulators of ENDO180. Plasma membranes are prepared from cells expressing ENDO180 by standard methods. The membranes are immobilized on wheat germ agglutinin-coated scintillation proximity beads (Amersham). Test sample and radiolabeled ligand, which is 3H-mannose are added and bead-bound radioactivity is measured. The ability of a given sample to modulate binding activity is determined by comparison of bead-bound radioactivity in the presence of the compound with values obtained in solvent controls. The test compound optimally produces a result that differs by at least 3 standard deviations from a set of at least 30 replicate control samples.


Example 17
Screening Assay to Detect Modulators of CDK10

The following is an exemplary assay for finding modulators of CDK10. CDK10 may be screened using, for example, the non-radioactive IMAP system available from Molecular Devices, Sunnyvale Calif. 94089, according to the manufacturers instructions. The ability of a given compound to modulate activity of the kinase is determined by comparison of fluorescence polarization values in the IMAP assay in the presence of the compound with values obtained in solvent controls. Preferably, the test compound should produce a result that differs by at least 3 standard deviations from a set of at least 30 replicate control samples. More specifically, IMAP is a technology that uses the specific binding of metal (M III) coordination complexes to phosphate groups at high salt concentration and a fluorescence polarization readout. In a microwell assay format, fluorescently labeled peptides are phosphorylated in a kinase reaction. After the reaction, proprietary IMAP nanoparticles derivatized with metal (MIII) coordination complexes are added to the assay to bind the phosphorylated peptides. The binding causes a change in the motion of the peptide, and results in an increase in the observed fluorescence polarization. This assay, unlike antibody-based homogeneous kinase assays, is applicable to a wide variety of tyrosine and serine/threonine kinases. IMAP technology is largely independent of the sequence of the substrate peptides. In addition, the technology is useful in assays of phosphodiesterases and phosphatases. In a typical reaction on a 384-well plate the following would be combined: 10 ul of the kinase in question, 5 ul of test compound, 5 ul of substrate, such as Myelin Basic Protein Fragment 4-14 (MDL Number MFCD00133371; CAS Number 126768-94-3) or a similarly-sized peptide, at a concentration of 30-1000 nM; and ATP at a concentration of 1-30 ul. The substrate may be labeled with a fluorophore such as fluorescein-5-isothiocyanate (FITC Isomer I). These reagents would be allowed to react at room temperature for one hour. Then 60 ul of IMAP binding reagent is added and the fluorescence polarization is read.


Example 18
Screening Assay to Detect Modulators of TNNI3K (CARK)

The following is an exemplary assay for finding modulators of TNNI3K (CARK). TNNI3K (CARK) may be screened using, for example, the non-radioactive IMAP system available from Molecular Devices, Sunnyvale Calif. 94089, according to the manufacturers instructions. The ability of a given compound to modulate activity of the kinase is determined by comparison of fluorescence polarization values in the IMAP assay in the presence of the compound with values obtained in solvent controls. Preferably, the test compound should produce a result that differs by at least 3 standard deviations from a set of at least 30 replicate control samples. More specifically, IMAP is a technology that uses the specific binding of metal (MIII) coordination complexes to phosphate groups at high salt concentration and a fluorescence polarization readout. In a microwell assay format, fluorescently labeled peptides are phosphorylated in a kinase reaction. After the reaction, proprietary IMAP nanoparticles derivatized with metal (MIII) coordination complexes are added to the assay to bind the phosphorylated peptides. The binding causes a change in the motion of the peptide, and results in an increase in the observed fluorescence polarization. This assay, unlike antibody-based homogeneous kinase assays, is applicable to a wide variety of tyrosine and serine/threonine kinases. IMAP technology is largely independent of the sequence of the substrate peptides. In addition, the technology is useful in assays of phosphodiesterases and phosphatases. In a typical reaction on a 384-well plate the following would be combined: 10 ul of the kinase in question, 5 ul of test compound, 5 ul of substrate, such as Myelin Basic Protein Fragment 4-14 (MDL Number MFCD00133371; CAS Number 126768-94-3) or a similarly-sized peptide, at a concentration of 30-1000 nM; and ATP at a concentration of 1-30 ul. The substrate may be labeled with a fluorophore such as fluorescein-5-isothiocyanate (FITC Isomer I). These reagents would be allowed to react at room temperature for one hour. Then 60 ul of IMAP binding reagent is added and the fluorescence polarization is read.


Example 19
Screening Assay to Detect Modulators of FPGT

The following is an exemplary assay for finding modulators of FPGT. Incubate enzyme plus test sample plus substrates (fucose-1-phosphate and alpha-33P GTP (Amersham)) under conditions described (Pastuszak, I., Ketchum, C., Hermanson, G., Sjoberg, E. J., Drake, R. and Elbein, A. D. GDP-L-fucose pyrophosphorylase. Purification, cDNA cloning, and properties of the enzyme J. Biol. Chem. 273 (46), 30165-30174 (1998)). Add to the reaction mixture streptavidin coated scintillation proximity beads (Amersham) precoated with biotinylated lotus lectin (Vector Labs, Burlingame Calif. 94010). Measure bead-associated radioactivity by scintillation counting. The ability of a given compound to modulate activity of the enzyme is determined by comparison of bead-bound radioactivity in the presence of the compound with values obtained in solvent controls. Preferably, the test compound should produce a result that differs by at least 3 standard deviations from a set of at least 30 replicate control samples.


Example 20
In Vitro Production of Target Polypeptides

Target polypeptides encoded by the polynucleotides described herein may be produced by the methods described hereafter. cDNA is cloned into a pIVEX 2.3-MCS vector (Roche Biochem) using a directional cloning method. A cDNA insert is prepared using PCR with forward and reverse primers having 5′ restriction site tags (in frame) and 5-6 additional nucleotides in addition to 3′ gene-specific portions, the latter of which is typically about twenty to about twenty-five base pairs in length. A Sal I restriction site is introduced by the forward primer and a Sma I restriction site is introduced by the reverse primer. The ends of PCR products are cut with the corresponding restriction enzymes (i.e., Sal I and Sma I) and the products are gel-purified. The pIVEX 2.3-MCS vector is linearized using the same restriction enzymes, and the fragment with the correct sized fragment is isolated by gel-purification. Purified PCR product is ligated into the linearized pIVEX 2.3-MCS vector and E. coli cells transformed for plasmid amplification. The newly constructed expression vector is verified by restriction mapping and used for protein production.



E. coli lysate is reconstituted with 0.25 ml of Reconstitution Buffer, the Reaction Mix is reconstituted with 0.8 ml of Reconstitution Buffer; the Feeding Mix is reconstituted with 10.5 ml of Reconstitution Buffer; and the Energy Mix is reconstituted with 0.6 ml of Reconstitution Buffer. 0.5 ml of the Energy Mix was added to the Feeding Mix to obtain the Feeding Solution. 0.75 ml of Reaction Mix, 50 μl of Energy Mix, and 10 μg of the template DNA is added to the E. coli lysate.


Using the reaction device (Roche Biochem), 1 ml of the Reaction Solution is loaded into the reaction compartment. The reaction device is turned upside-down and 10 ml of the Feeding Solution is loaded into the feeding compartment. All lids are closed and the reaction device is loaded into the RTS500 instrument. The instrument is run at 30° C. for 24 hours with a stir bar speed of 150 rpm. The pIVEX 2.3 MCS vector includes a nucleotide sequence that encodes six consecutive histidine amino acids on the C-terminal end of the target polypeptide for the purpose of protein purification. target polypeptide is purified by contacting the contents of reaction device with resin modified with Ni2+ ions. target polypeptide is eluted from the resin with a solution containing free Ni2+ ions.


Example 21
Cellular Production of Target Polypeptides

Nucleic acids are cloned into DNA plasmids having phage recombination cites and target polypeptides are expressed therefrom in a variety of host cells. Alpha phage genomic DNA contains short sequences known as attP sites, and E. coli genomic DNA contains unique, short sequences known as attB sites. These regions share homology, allowing for integration of phage DNA into E. coli via directional, site-specific recombination using the phage protein Int and the E. coli protein IHF. Integration produces two new att sites, L and R, which flank the inserted prophage DNA. Phage excision from E. coli genomic DNA can also be accomplished using these two proteins with the addition of a second phage protein, Xis. DNA vectors have been produced where the integration/excision process is modified to allow for the directional integration or excision of a target DNA fragment into a backbone vector in a rapid in vitro reaction (Gateway™ Technology (Invitrogen, Inc.)).


A first step is to transfer the nucleic acid insert into a shuttle vector that contains attL sites surrounding the negative selection gene, ccdB (e.g. pENTER vector, Invitrogen, Inc.). This transfer process is accomplished by digesting the nucleic acid from a DNA vector used for sequencing, and to ligate it into the multicloning site of the shuttle vector, which will place it between the two attL sites while removing the negative selection gene ccdB. A second method is to amplify the nucleic acid by the polymerase chain reaction (PCR) with primers containing attB sites. The amplified fragment then is integrated into the shuttle vector using Int and IHF. A third method is to utilize a topoisomerase-mediated process, in which the nucleic acid is amplified via PCR using gene-specific primers with the 5′ upstream primer containing an additional CACC sequence (e.g., TOPO® expression kit (Invitrogen, Inc.)). In conjunction with Topoisomerase I, the PCR amplified fragment can be cloned into the shuttle vector via the attL sites in the correct orientation.


Once the nucleic acid is transferred into the shuttle vector, it can be cloned into an expression vector having attR sites. Several vectors containing attR sites for expression of target polypeptide as a native polypeptide, N-fusion polypeptide, and C-fusion polypeptides are commercially available (e.g., pDEST (Invitrogen, Inc.)), and any vector can be converted into an expression vector for receiving a nucleic acid from the shuttle vector by introducing an insert having an attR site flanked by an antibiotic resistant gene for selection using the standard methods described above. Transfer of the nucleic acid from the shuttle vector is accomplished by directional recombination using Int, IHF, and Xis (LR clonase). Then the desired sequence can be transferred to an expression vector by carrying out a one hour incubation at room temperature with Int, IHF, and Xis, a ten minute incubation at 37° C. with proteinase K, transforming bacteria and allowing expression for one hour, and then plating on selective media. Generally, 90% cloning efficiency is achieved by this method. Examples of expression vectors are pDEST 14 bacterial expression vector with att7 promoter, pDEST 15 bacterial expression vector with a T7 promoter and a N-terminal GST tag, pDEST 17 bacterial vector with a T7 promoter and a N-terminal polyhistidine affinity tag, and pDEST 12.2 mammalian expression vector with a CMV promoter and neo resistance gene. These expression vectors or others like them are transformed or transfected into cells for expression of the target polypeptide or polypeptide variants. These expression vectors are often transfected, for example, into murine-transformed cell lines (e.g. adipocyte cell line 3T3-L1 (ATCC), human embryonic kidney cell line 293, and rat cardiomyocyte cell line H9C2).


Provided hereafter are NRP1, NID2, ENDO180, CDK10, FPGT, PCLO and REPS2 genomic sequences (SEQ ID NO: 1, 2, 3, 4, 5, 6 and 7, respectively). Polymorphic variants are designated in IUPAC format. The following nucleotide representations are used throughout the specification and figures: “A” or “a” is adenosine, adenine, or adenylic acid; “C” or “c” is cytidine, cytosine, or cytidylic acid; “G” or “g” is guanosine, guanine, or guanylic acid; “T” or “t” is thymidine, thymine, or thymidylic acid; and “I” or “i” is inosine, hypoxanthine, or inosinic acid. SNPs are designated by the following convention: “R” represents A or G, “M” represents A or C; “W” represents A or T; “Y” represents C or T; “S” represents C or G; “K” represents G or T; “V” represents A, C or G; “H” represents A, C, or T; “D” represents A, G, or T; “B” represents C, G, or T; and “N” represents A, G, C, or T.


Following is a NRP1 genomic nucleotide sequence (SEQ ID NO: 1).

>10:33587901-338095001gcaatgggtctcaagcaaagcattcttcaagtgccccacagttatcaaataatttaaaaa61tcttagattttcatttttgtgattatcttctaaaaatggcagttatacccattgtggagc121acctagacaggtaccatataaggagtactgccatgtctttttgtagcaacacatatgttt181gtgttgatgattgaatgactgccaagtgcggtgataggaaagcatgaatccctataYagc241caaggttttccagttgttcaaaaataaaacagtggtgggagaggtgagccatcacttggg301aacacaggagtaacatggatctgaactcgaaagaccccgcatcctggcctttggcagcat361gttcaagtctgtggacaattgataagcaacagttcattcgtggtcagtctagacataaat421aggcagtgaagaactttccattagaagattttcttcccagaccaggtgcagtggctcacg481cctgtaatctcagcattttgggaggctgaggtgggaggactgcttgagtacaggagttcc541agaccagcctgggcaaaagagccagatcctgtctctacaaaacaatttttaagttagtca601ggcatggtggtgcacacctgtagtcccagctacttgggaggctgaggtgtgaggatcact661tgagcccaggaggttgaggctgcagtgagccatgatcatatcactgcactcagtgtgggc721aataaagtgaaacccagtctcaaaaaaacaaaagatttttctccctgtattatattttgg781gggtacaccattagatcatttggcctgttagtagataaaacaaattaaaagataaaaagc841aaatacataaaaataaacagcaaaggcaataatgatggctaaaagggctgcctattatgc901taaaactcaaggaaattaagttaataacaagcaaagcctaaagtgcaagtacgttctact961agtatattacccaataacaggaattttgcttctacaaaatgccatcagtcatcacaccaa1021attaatgttgtcatctgagtgtttgggttttgtagttatgttgggattagtgtgtaaagt1081tgcttggactcatttgttgtatgggaatactaagggtaaaaaaatgtgtgagtgaatttt1141ttgtaggtatataataagattagaataaaagagtgatcttcaacaccaaagatccttgga1201aatgtttccctttaaaagagggtcatacattccttaagttggggaaatactgtgttttct1261ttcagctccatcctcgggcctttcagtgaggacaggaacccacggccagacaggaaggcc1321tgtttgaagccaggttgcctccctagtaagtttgttctggagccttctggtctccttttc1381ttttagttactttaagtggttttaggaaaaagtactcttttcatcttaatcctgctcctt1441acacccaagccagcccagagccatctggaatcctcagaacagacctattctttgaagaga1501agaaggacaagatgtttttggacccttgcaggaggtctggatacagacggggccctgggg1561agggttgatccccagcagtaactgctcagctggaggcatgaggggtcccttctgacctcc1621aagcttcagaggggatgggaaaaggtaatttggcagctttgtgttccaaggcttcacccc1681tagattggtttaggcaagagtcagtgagtccctgggaaggcactgggccagcagagccca1741gacatcaaggctagcatttcagtcttaataatccaatttaggctgggcgtggttgctcat1801gcctgtaatcccagcaatttgggaggctgaggcagatggatcatctgaggccaggagttc1861aagaccagtatggccaacatggcgaaaccccgcctctactaaaaatacaaaaattagccg1921agcatggtggtgcatgcctgtagtcccagctacttgagaggcttaagcaggagaatcact1981tgaacccaggaggcggagtttgcagtcagccaagatcgtgccaaaagaaaaaaaaaagaa2041taaaaaaccaatttaaacaaagaaaaacctttcattgagctctttaaatatcctaagaat2101tgattataaatcacaaattggaagccaacataagctgtaaYgcaaagcaagcactgaaca2161tctctctacctgatgccacttaagttatggtaaatttagtcaaggctgtgggttggggtt2221gtgtgccttattgcaggaagcagcacacttctgagggataaatattgcatttggaaccta2281tgagttggtagtttttctccaaggatgttcttaaaagccatttgcaatgaatttgtcagt2341ttctttagatttggatggaatataacgattctggaacattactggggcctctgaaaattg2401tatttcctaccttatatcttacgtggctaaagtacatggctgggagcggtggctcacgcc2461tctcatcccagcaatttgggaagccaaggtgggtggatcacctgaagtcaggagtccaag2521accagcctggccaacatggtgaaaccctgtccctactaaaaatacaaaaattagctgggt2581ttggtggcatgtgcctgtagtcccagtctacttggaaggctgaggcaggagaattgcttg2641aacccaggaggcagaggttgcagtgagctgagactgtgccactgcactccagtctgggag2701acagagcgagactttgtctcaaaaaaaaaaaaaaaaaaaaagtacatcttgttccaagaa2761acctacttttccttcaaacactttgaggtttaaaggcaggatttactaccctctgtcctg2821acggcagagggagaaggaggagaggaaggaacagaggctgtcaccccttgatgattttct2881gggatcctacagttagatgggggtatggtctcctgccctatgaaaaccagattctaggat2941gctattctaagaaaaaaaaaNtcccctggaatcatattatcttctgaaaactagtgccac3001attttcctgttgatactatactcatcaacttcagttcacagtcaaaattcagttatcctg3061ttaatcttgctcattttggcaggcacgaaaatacacatgtttgtatggagccaagcaagt3121ttttgagatcttgggatcctctggaaatattttttcagttgttgaaaaggaacttgcaag3181aaagcatatattttcaacactcgtcccaggaacagtgtgtgaaagtgacattaaacttat3241ttctacactgggtgactagtgactccctgctgggtaatcaggtagacctctgtcatctct3301gcttcactggggtttactgccagggcgtagagagtctcatctcatggatcggaagaatgg3361gctgtacacacagacagctctcaccctcctgttctggcgctgggctattcttcacggcca3421ctctttacactgagaaacacctcatgaattttcagcctggaagacgatgggcccatttaa3481tgttctaaccatatagagtgcctgaattcccaagatctccaactgcttttcacttactac3541cctggaaagaaccccagtagcatccgttatgaagacaagaactccgcaaatctcctcctt3601gctggctggggagcaggaggtctcataattctgtatctggccaggcatcgtccttggctg3661catgttggttttggcatcagaggcagctgcctctaagcccttgttctgccatttcaagtt3721atgtattcttaggcagatcttttcatctttttatacttagtttcctcctttgtaagatga3781gaaagatagtatcttgcagtgtacttgtggcgattaaagattatatttacacgcatacca3841ccaggcacaccgtagacaccacacgaattctagctgtcatattccaggcctaggctagaa3901tgttgtcagaataaaagtttcatattcttcccactaatcaaaatactctcaccaccacca3961ctactgctattactacttctacctaccatgcattgagcacttatttgtgctgggaacttc4021actaggtactttgctgcataagcttggtttgttttatactttaaggaccccacaaggtag4081gcactatttttagccccacttggtcactgagataaatgaggcatgatgaggttacattgc4141tcaaggtcacagaggaagtgtctaaccctggctctgtctgacccaaagttactgatctga4201aaacgatgctacgccttttctagaaaatgagccaaacctgcaccccaccctacaaagcag4261tccctgcccccctgaccctgccccaatccttagaactacctatcttctttgtattcatag4321ctcttccaagcaggagtccatcagcactgcaggctgagcacaacataccagcttttgggt4381cagacaccagctgctgaaacaggcccttcaatctgttgtactccccagaattgcctctga4441tccttgcctgacaaccatcccaggatttcagatcaagatgtctgaggcaggcccacttct4501aggtattttttgaaaaaatttgttacaggtgacagggcgtggtggctcacacctgtaatg4561ccagcactttgagaggctgaggtgggaggattgcttgcggccaggaggtcaagaccagcc4621tgggcaacatagtgagaccccagtctctacaaacaaataaaatttaaaaatcggctggtt4681tta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atgtgtgaatttgatcctgtcattatgatgttagctggtgattttgctcattagttga219961tgcagtttcttcctagtctcgatggtctttacattttggcatgattttgcagtggctggt220021accggttgttcctttccatgtttagcgcttccttcaggagctcttttagggcagtcctgg220081tggtgacaaaatctctcagcatttgcttgtctataaagtattttatttctccttcactta220141tgaagcttagtttggctggatatgaaattctgggttgaaaattcttttctttaagaatgt220201tgaatattggcccccactctcttctggcttgtagggtttctgccgagagatccgctgtta220261gtctgatgggctttcctttgagggtaacccgacctttctctctggctgcctttaacattt220321tttccttcatttcaactttggtgaatctgacaattatgtgtcttggagttgctcttctcg220381aggagtatctttgtggcgttctctgtatttcctgaatctgaacgttggcctgccttgcta220441gattggggaagttctcctggataatatcctgcagagtgttttccaacttggttccattct220501ccacatcactttcaggtacaccaatcagacgtagatttggtcttttcacatagtcccata220561tttcttggaggctttgctcatttctttttattcttttttctctaaacttcccttctcgct220621tcatttcattcatttcatcttccattgctgataccctttcttccagttgatcgcatcggc220681tcctgaggcttctgcattcttcacgtagttctcgagccttggttttcagctccatcagct220741cctttaagcacttctctgtattggttattctagttatacattcttctaaatttttttcaa220801agttttcaacttctttgcctttggtttgaatgtcctcccgtagctcagagtaatttgatc220861gtctgaagccttcttctctcagctcgtcaaaatcattctccatccagctttgttctgttg220921ctggtgaggaactgcgttcctttggaggaggagaggcgctctgcgttttagagtttccag220981tYtttctgttctgttttttccccatctttgtggttttctctRcttttggtctttgatgat221041ggtgatgtacagatgggttttcggtatagatgtcctttctggttgttagttttccttcta221101acagacaggaccctcagctgcaggtctgttggaataccctgccgtgtgaggtgtcagtgt221161gcccctgctggggggtgcctcccagttaggctgctcgggggtcaggggtcagggacccac221221ttgaggaggcagtctgcccgttctcagatctccagctgcgtgctgggagaaccactgctc221281tcttcaaagctgtcagacagggacacttaagtctgcagaggttactgctgtctttttgtt221341tgtctgtgccctgcccccaRaggtggagcctacagaggcaggcaggcctccttgagctgt221401ggtgggctccacccagttggagcttcccggctgctttgtttacctaagcaagcgtgggca221461atggcgggcgcccctcccccagcctcgttgccgccttgcagtttgatctcagactgctgt221521gctagcagtcagcgagactccgtgggcgtaggaccctctgagccaggtgtggaatatagt221581ctcgtggtgcgccatttctt


Following is a NID2 genomic nucleotide sequence (SEQ ID NO: 2).

>14:50462485-50525750rs3742536:32929rs75305:34983ctttttacctgttgggcagtaggggtagactgcagttatcccgtagaggtgagatcgttg60ttctgggagatactcatcagtaaactggccactatgtttatttactgatacaacaccatc120cctggtaacagaaaggaagagtgaattcagggacagtcatttactagataaagaagtcag180tcagccacagaaaatcagttgcaatagaggaaaatttctggcagccttctctgtcccagg240gctggtgctaaagccatactgaagtttgaagaccattgctctaaatccattgctcatctc300tagctgcatgttagaatcacatgatgagctatcaaatcagagttgtagggcatggtgtaa360agggcttagactttaatgttccacagttcctcatttgagaactagtctaaatagtatttt420tcggtccctcagacaatatgtccacagtgtcaaaagccaacttaagccctgtaatatagc480tcatggtatcaagggtagtcttattctctaaagaacatatttagattaatatgctccctt540ttgagacattatccaatagctttgctactttttaagctatatgtgagcaataccattcga600tttctgggtgaagtaaaaggatgatttcaaaaacatgaaaggatgaaaaacatccttgaa660ggatcttatctgccaatgaggaggtgatgaaacaaaagcctcagagggcttaatgagtca720agtcaggtactgacttttggtaaaacaagtggtgtgtcctctggaacagttggccttttc780aggcttgtccagaatttgtggggctcacctcctccagtctgtgtggtagaagtgatctgc840atagcttacgatgctgaaggggtacttgaggttgttttgaatgacacgccgtccagttcc900atcaggtagtgtacactccagttttttggttccttttaaaacaaagggggaaaatgaggt960ccttcagctgctttgccaaaaatgatagtgacatagtgatagtgacacagtgatagaatg1020gggaaataggatattttacttccattacagtcatagattagcaactgtaaaattacctgt1080cctattactagtctccagtttttagtagagattcaaactttcaatccttttttggaagac1140aggattgcatttactgaaatctggtaagtttctgccaaagtagggcattaaacataatta1200tttttataatttgtgtgatttcaagccttagcttataaattataacatctgggtaaatac1260ttgccctttgtaagctatgtctataattactgaggcaaccttttaaccaaaatccttagt1320atttttaaaatattttctaggtaggaggaaaaaagtgactaaaatggtaacataactcat1380atttcaatgtaaaggaatcagtcttaaaaatttttacaatagctaaaccattcaagaatt1440tccaagtttcacatttctcatggagccgatttaaatttcaaatccttttaagaaatgtaa1500cttgttttaaacaactgggttataaaagtgacctatgtacattaggcccttttattagat1560tacaaattcaagaacagtctttttcatattctttagtatagaactagacacacagcattt1620tttcagttgtgctgataaaagcaaacaaatgtaggttgctaatacccatgttattactga1680aggaagctaccctgctacaatgctttccaaagaatgtccttagcattataacagatgttt1740ataggtaagttatagtgaccctctccccgcataagaataacttcacttaagtttgtcaat1800tcaacaatggctaattcttttaactgttaaaaatatgccaggcactcatggtcaggcaag1860cagtacaaacttactgcttgatatatccttccctccacccaaaccccagaaagttcattt1920taagactcatttactagtacttaaatttactagtacttaaaagtttacagaccaaagaaa1980ggagatctaagtgatgggatttcattttgtagtaaaatctgttagtgctcacattcactg2040tgatgagaggttatctatttgcattccatcagtagtattacctgcatctgcccagcagag2100cagtttagagaaagggtcaaaggttaagccattgggcaatccaatgtctgtattgatcag2160aattcttctgttttctccatctaaagatgacgtttcaattttaggagcttctctattcca2220gtctgtccagtacaagttgctgtaagttaaaaatcaagattgtaaaagaatagccatgta2280gcctgtggtaggcagcatctaagcagcccccagtgatctccatctcctcatgtattatag2340ccttaagcaatccccttgagtttgggctgcattagtagtgtcttgcttcttctagtgcat2400agagtatagcagaagtgataggctatcactgccgatattcggtctcaaaaaactggtttc2460tgccttaggggctctctcttgctccctttgagggaagctggatgccatgttgcaagctac2520cctgtaaaggggaacatgtgccaaggaactgatgtccttgagggctgttggccagagaca2580ttagtcttgactgtagcccatgtgagcatgcttgggctgtgtcttccccgagctgagcct2640tgtgatcactgcagtttggccattacctgattgcagccttgtgagagactccatgcctga2700gggagctcggtgaagacacacttggattcctgactcacagaaactgtgagactaaatgtt2760atcttacactgctgagggtttgggcaatttcttacacagcagtaggtaactaatacacag2820ccatattccacttttacctgtatcacaaggttttctgattattgtcactatgtgggagaa2880gtcattactaagttactatcagaatccctgcctccaagataccccttgactcaactgccc2940caaaaccctccgtttgttgaagtcttgaattggatcttgggttttagaatagaaacaaat3000acagctgttaggaatggcttcgatacaaactatggaacttgaagagatgtggaccaggga3060tttgagaaggaaactgttctttcttctgccaaaaatacaatttggtacttgaaggggtac3120ctggcacccagggatggaaacattttaagttaagcactttaccttttaaatgccacgtta3180aaaatgaaatatattcataacagatactaacgaggaaatatggtcctattaagtggaaat3240aggttataaggaaacaaacaggtttatcgttagagcagaaataatgttgaaagcactttt3300tataaaatactattttataaaagctatagaagctttgaataaatcagttgggctacagag3360aacctcagtgagaggagaggaagaaaccataggtttcccctgttgcgaatttcatccctt3420atgcccaacagatgaagcagagagaagctgagagcaaaagctagaaagcagtttccactt3480tgcgctaaggtggaggtggccagagggtacagcatgcatatcattgcttcatcgcttctg3540atcagtctcctccccaaaatgatttaatatcatcaacaacaataagtctgaaacaagctc3600acacagatatttggtgtaaattatttggggattgccagaggaagaaagttgtaaaggtga3660aatctatagaaagcaaatgagaaatatgcaggaaggcttgacaaccattctgttacgcag3720ttgaaggagatgtaattacatgtgttatattgcaacctggttcaaggtgggttttctttt3780tattaaaaattcaagtcctttagcatttatatttgctgtctggataagaaagttctactt3840tcatatttaaactcatggctgtgatgtggatttcagcttaaaagtatgctttttccccag3900actccagagacatggccaaagtttcattgcctacaagatggcagctctccccagttttca3960ggtcaagcaccaagaaggaataaggggccgggcgcagtggctcatacctataatcccagc4020actttgggaggccaaggcaggtggatcacccagggtcaggagttggagaccaaaatggcg4080aaaccctgtctcgactaaaaatacaaaaattagctgggcatggtggcacatgcctgtagt4140cccagctacttgggaggctgaggcaggagaattgcttgaacccgggagatggaggttgta4200gtcagccgagactgtgccactgcaccccagcctcggcaacaaagcaagaccttgtctcaa4260aaaaaagaaaaaaaaaagaaaaaaggaccttccaaatgtccaaatgtcagaaaggcaagc4320ctatgcaaagagtttttaagagggctattaattcctgttaagtcaactggaatagagaag4380gatacaggtacttttcctgtgtgtagaacccaaagtttgaaaccacctttgaggcactgc4440actaagtatttcatagtcaaaaagcacttacctgaaaattaaagctgaaggttttcttta4500ctgatttgaagggcatctgggaatttcatttcgtagaaactccacttaaacatggcaaga4560gtttctcaagatataaaaattctccctgattgtaacaagtactgagacagtgaacaaatc4620ctgtatgtcacatttgatgaccttagcaattatacagaca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aaaacaaaaagcaagaaatgtttttagagcttctcaatactaggaagccacaactactgc56700 caaatataaacaaaggctgttctgcctcatgtctatccataagatagtcgacatcatcaa56760 ctaatgtggactcaataacttcagagttatacattagtcagaataaagaaaaatcaacta56820 gggaaagaaaagtactagaggaaggagaaaaaaccccatttcaaaaagctaagctttcaa56880 agctcctggcattgatctgccataataaatgtgagataggagagaatcctggttctaaat56940 ttagagaaattccataagataaggaggttgtagagactgtgaagaattgaaggggatggg57000 gaaggggggaagagagtagtaaacagagctgtgaagaggagagtctgactttttgctgtg57060 aactctgggcatttagcactttattttattcattcattcattcaaccagccaacaaattc57120 atttcaattaaaatttattcaacatcttctatatactgttcttgagatgagcactcatta57180 tgggtggcaagtatttaatttctttgtctccaccaaaataggcaaatatcagaggaagga57240 aacaacttccttttaagtagaatttatgatcccaaagtacacgccaaaagagtttggctg57300 tttaaaagccacccacacactaaatgatcacttgtaaagttcttggtcagttctacaaaa57360 ttctacatctaattgctaaagagcccacttcatgaagtcttgaagggctccccctctcaa57420 aaaactccttgactggcttccaaacaggaaaggtcctcacattcctagatacaagctttt57480 gcaagagccgcacaggctctgtctccaaagagaagccctcagatgggctgccaaacctga57540 caaacgagtcactggttcccagctcccccaggatcccagagaggagagaggggcctccac57600 agggagaggagcagcacagagaggatcagggtgagtggctgcaggccctgttatcagaaa57660 gagagaaaagccagagccctctgcagaaaccagagcctgactctccctcccaggtacagg57720 acatcccaatcaatgttgaagtgacactcatcagagccaggctgatgcatgcaacataca57780 cccctctaagagtttctaattttttcaattcatttaattaactgcagctatctggacaaa57840 aatcagaatagtgacatccttttcttcaaggggcttgcctatagaggcttatcaaaatga57900 aaatgaaaatggaaatgaagaattagggtgactcttataggggtcacaaagttctctttg57960 tcatcttcctcaggaccacatgtcctctagagctctaggatcattgggtctggcctccag58020 tgtttgctgtgaacactaacctccattccttagtgtagttcttcatctcccactgcctgt58080 aggacactgtcacatgggggactgtccctcatttattttactgcactaaatcctacagat58140 cctgccaaaaattcaaagacaaatgctgagtgctatcacatccctaatactatatccttc58200 ctgatactcttcagcaaaacgtcagcatttctaatttccatatatggttcagcattaacc58260 aatttccatatatgtctctattctcttcatcttcacagtaatatctcagactcagtctga58320 actctcttggatgggctgtagaccacccctcttctaaaccttgtctccagaagtcaaaaa58380 gctacaataaagtcctgcttttacctctcattttctctgtgaaatccaaccacctccatc58440 aatgactatcaacctttgaaccaagtcaacatgtctcagtctccacccagatggaggcct58500 ccattgggcagtgtcttaatttgtcttctgttgtttacaacaaaatacttgaaattgggt58560 aattcacaaagaaaaggaatttacttcttacaattatggaggctgagaagtccaaagttg58620 agaggtcacatctggtgagggccttctggctagtggtgactgcagagtgccagggtactt58680acggtatcacatggcgacaaagctgagtgtgctaatgtgctagctcaggtctcaattcct58740cttcttataaagcaaccaggcccactccatgataatccagtaatccatgaatgaataaat58800taatccattcctgagggcagagccctcatgacccagtcacctttcaaaggccccacctct58860caatactgtcacattgaggattaagtttcaacatgattttcagaggggacaaatattcaa58920ccatagcgggcaaccttgcactcatctgtcttatgttactgcctgtgactccttcatcca58980tttctctggcccctttgctaataggttgctctctcaaccttctcctgggtcatgtttttg59040ctcagaacaatctctcttgatacctcattctcaatatccattctttttagccctttcaac59100ctgacttaaattccagataaacaaaacattccccacattacagagatctatataactcag59160ctgtctctaaccccatccctaaaatgacaaaaacaaaacactcctctttccccacaaaaa59220gcacagctaaacattagtcctctatgtctcccatcaataagtatttcaaaggtctacacc59280ataacttttttgctctgtgaactacctatgagagttcaagaaatctcagattaactgaat59340tgacaaacactatttgttcacatttttttttcagatacaattacaccttgacattcctgc59400aatatcaggtgcaacatctggtcacacctgacaggaatatccatggagaaaaagggcacc59460aactgcaattgtttttaatggcaaagatgccattgctttgaatcttccagtttccagtaa59520ctttgcaatataaagttagtaagccaaactttaagaacgccatgtagaactcagtccttc59580aagcaaattagattactgtttgccaaagtctatctccaaagaactattaaaacaagaact59640caagtcccagagctctttaaaatattggaagaaacttcaatgtgggtaattgaaagaaca59700tcccctatccaaacagaatcctcctttctttttttttttttttcccctttcttttttatt59760tatttatttatttatttatttattttttattatactctaagttttagggtacatgtgcac59820attgtgcaggttagttacatatgtatacatgtgccatgctggtgcgctgcacccactaac59880gtgtcatctagcattaggtatatctcccaatgctatccctcccccctcccccaaccccac59940cacagtccccagagtgtgatattccccttcctgtgtccatgtgatctcattgttcaattc60000ccacctatgagtgagaatatgcagtgtttggttttttgttcttgcgatagtttactgaga60060atgatggtttccaatttcatccatgtccctacaaaggacatgaactcatcattttttaag60120gctgcatagtattccatggtgtatatgtgccacattttcttaatccagtctatcattgtt60180ggacatttgggttggttccaagtctttgctattgtgaatagtgccgcaataaacatacgt60240gtgcatgtgtctttatagcagcatgatttatagtcctttgggtatatacccagtaatggg60300atggctgggtcaaatggtatttctagttttcttaatgaaacattctgaagccaactagca60360aatggtaatgtctgcaggtggattttaattagaaaagtaaaagagagatggaatttacaa60420atctcttctaccctaaacctcccccaccccagaaaaggaccagacaaaagttttaattcc60480aaaaaccttgcatttaaagagtccttatgcaacatagagagtaacaggatggttaccaga60540ggctgggaagggtagtaggggtcaggggaaggtggacacagttaacgggtaccaaaaaaa60600aaaaaatagaaggaatgaataagacctaccatttgatagcacaacagggtgactataacc60660tgtgataacaggttattatagtcaataataacttaattgtacatttttaaataactaaga60720gtgtaattagattgtgtgtaatacaaaggataaatgcttggggggatggataccccattc60780tccaagatgtgattattacgtattgcatgcctgtaccaaaacatcccatgtaccccataa60840atatatacacctactatgtaccctcaaaaattaaaaaataaagagtccccatacaaaaga60900tataagtcacagcatccttacaagatagagatcagtgacatgggcaagcaacctcacctc60960ttacagggggcatgtaccatccttctatagtgaagaagcatttccgtaagttaaaattga61020gatgaatcatatggtttcagcccctagagtatggtattttagattcaggtctaattccat61080agaaggaagcccttactttcttcaggaatgaggcagtttgccatttaccaagcacgctgg61140ccccactcatatccatttggaggacttcaaatgttgatgacaagactaaagaccaggagc61200ctggaattttttactttccgcaggtagagactgtaaagcacattgacggcagactcctat61260aaactaagaaacttgtaaactggtacctggaaggcaattaggcagaagttatcaagaact61320ttatcaaaagtgcatcctctctgacctaacaattgcacttcctggaatgtcaactaagaa61380catgtgcaaagatttagctttttaaaaagtgattcacgctattatttataataatgaaaa61440accagaaaaagccttaatggcccaacactggttgagcatactgtggtacacccactatta61500tacaggataatagcaggatacagtagtatgcatcattataaatgatgtagaaatacattt61560attgacattgaaacatatttacaatacactattctgtgaaaaaagcaagttataaaacag61620tatgtgcagtgtaaactcatcattgttaatagacagcaataaagatatgcagaagaaaaa61680actctgaaaatgtacgcagaaaaatgtttatcagtggtgggaaaatgagtggttgttttc61740ttttttaaaattatctgtattttctgattttttccccttcaatgaacatataattagaaa61800tacttgtatacttagagatggttctaaaaataagcaaccaaacagtaaattgccttatgt61860tcaaaggggctcccacatctgcactcccgctctcatatccgctaagcatttttttcctca61920agggagtgaccacctgccagggcctcagaagatgcctagagacaacgtctttcctcactc61980ccaagccactccagccctgctgcctccaaggtcaggttcagcgcaagagtaggctcagaa62040acaactccgagccagtccctgggtcgctgccccagaatttaagcccatccttcaatttcc62100tcagcagtcaaatatccctggtcacttacctctcccgagggcagcgccccgcgcttgacc62160tcctcgtaagcgcctacctgctcccaggtggccaggaaggcgtgggtgggggtaaagcgc62220gcagagcgcgggaagccagcgcgcacatagcgggcggccaggcccagcactgcgggggag62280gtgtcctctcggtacaggactcggcctctgccgtggctcgtgtcgatgtccgccagaaaa62340ggggcgatggccgggaagtcggtggggaaatcatagtccacatactgcgtttccctgggg62400aagtcctgagtggagatgatgccgttggtgcccacctgggagaggagagggacaaaaagg62460tgacagtcgctcaagcccaaggacgctaccctttcgcgcagggagggaagggaactgact62520taggcaaaaagcctctctctccttccccactggccagggagtctggattcagggaaacaa62580gcgcaacttttgcgactgtcccagctccgcgctgcagtaggtccaggggccctaaaactc62640gttcacctatcttctcactcccctcacaaccccggagccccatgtcctcggagaggaagc62700ctccaggtctgaagagggaacaccccagctccgcagacgggcacagccttctcgcggtcc62760taactgcaaagcccgctggccgccctagcaacgccaccttttccgcctgggacgcttacg62820tggctgcgaacaattcttgctctcccctccaggtcattcttgctcccctacagcagcggg62880gcaggcagcggcatcgctctctggccgggtaccaccgcaagggtgctggggggctccagg62940agtggggtctgtttcctcccctctggaggcagggtttccgtgagaccgaccccagggaag63000aagctgcgggaaaagtgccaggagggggctgaacttacgtagaggttgctgaatcgggct63060tcgtagaagtgcaggggattcgccagcttcaccacggctgagctttcgtcgtcgccttcc63120tgcaggagctggtccccccacgactccccgtgtgggaagagctcgtctgggtgcagcgcc63180gcggcccgcaacattagcaacggcagcagcagtagcactggtaacgacgacagcaccggc63240cgcccggccacccggtccccctccat63266


Following is an ENDO180 genomic nucleotide sequence (SEQ ID NO: 3).

>17:61200451-612686501catattgtgctgtgcttcaaatgtgtctcatgtctcctatgcagcgtgtcacagtgtcat61ttgacatcaatcatgttggcagggcagcagaccccccccgccccaccgatacatacattt121tttgttttgttttattttgaaatagatacagccttcttgtcattcttttttttttttaat181tttttttaaaatttttttgagacaagagtgtcgctctattgctKaggttggagtggcact241atctcggctcactgcaacctccgcctccagggttcaagtgattctcctgcctcagcctcc301tgagtagctgggactacaggcacccgccaccacacccggctaagttttgtatttttagta361gagacggtgtctcaccatgttggccaggctggcctcgaactcttggcctcaagtgatctg421cctgcctctgcctcccaaaatgctgggatcacaggcatgagccaccatgcttggccgtca481ttcttaatagacaccaggatactgaaaaatgctttgccactcagaaaaacaagggtacaa541gcatgatactatatggccactctcattccacttcagtgtgtgtgtatggtgggggcacaa601aagggatgggtggagactgtggcaaaccggtgagcccacacccctcccaggtgggcagct661gcaccagctcttgtggctggcaccctgcaggcatgctgcctcgactggaaatctacattt721ttatgtgtaatctctctcacacatcctgaagccaaacacaacttgcctgtgggctatcat781ctaaaattcggaggctagctggagtccgctctggaataagtctgcctggccatccttccc841agctcttccccttattaactgtgttatcaagatgttaatctctcagttatttaatacctc901agtttcctcatctgtcaagtgcagatcagaattttgcctacatctgagggttgttataag961gattaatgaaaagccttggaacccactgtctggtgcctggacgcgcatggttaatgcctg1021gcattctttgtgtctctttttacaagcgcagggtgaccacaggtcccagtttgccggggg1081caggcctggttcatgcttgtggtgctggtgtaatgatcactaacttgccatttcattgtc1141ggtattgacgtaaatgatatgatttgcctagagaagcaaatactggcacagtgctgttta1201ggactctgatgaggtttagacctaatagaaaggccatggtccaggttccttgaacatccc1261tggacaagtcacttaacctttctgtgttcagtttcccgttctaaacgattggcaagatga1321attctaagatccctccattcacagtcattctgtcatttgacaaacatctttttttttttt1381tttttaaacgcagtctcgctctgtcgccaggctggagtgcagtagtgcgatctcggctca1441ctgcaacctccgcctcctgggttcaagcaattcttctgcctcagcctcccaagtagctgg1501gactacaggcgaacgccaccacgcccagcaaatttttgtacttttagtagagacagggtt1561tcaccatgttggccagaatggtctctatctcttgacctcgtgatccacccacctcggcct1621cccagagtgctgggattacaggcgtgagccactgcgcccggccgacaaacatctttcaca1681agctcagactcatgccaggcaaaggtccagggcatggagaagccaggggcagagcctggg1741tcctcagtgggcacttggccagtgaaaacctccagtgcaggttgttgggaatctctacct1801tttttgtcaagtgttacttgaaagaccattaccaagaaattgaattgaaagttaattttt1861ttctccttctttaYggagacgagtttcactctgtcccccaggctagagtgcagtggtgcg1921atctgggctcactgtaacctcctcctcccgggttcaagcaattctcctgcctcagtctcc1981tcaggcatacaccaccacatctgccaccatacccagctaatttttgtatttttattagag2041atggagtttcaccatgttggccaggctggtctcaaactcctggccttaagtgatccacct2101gcctcggcctcccaaaatgctggaattaaaggcgtgagccaccgcgccctgcctgactgt2161taatattttaaacctcagtttctcaggaactaatcattctggttgaatacatttYgctac2221acatttcttttgtttgtgggggggtatctaagttgtttttttcttttaagtcagaaaaca2281taaattttcactatggaaaattgtaaattacagaaaggcccaaagattttcttcatttct2341gctgactcttcaatgaaccagaggggcacgtgtaatatttccgctttcggttactcccca2401ccccacacaccttgtagacactttgtcaaaactacattttaaaaaaatgctttcccccaa2461attcgtatggatgtctcctaataaagtcacgcagtcactcactggttcgtcctcaatcaa2521ccaggcgtggagtcctaacctgtccatgattttaagaaagggcaggcgcacaacaccccc2581gcacggagaaatgaacaggtgtggctcggcagaggccttccgcacctgcgttatggagtg2641gtggccgctgtcggcgatggggtccccgcgagggcacaagtgatcgcgtggccacggcga2701ggctctgcctccgcccccgcgctccgggtcccattcccgaattctgccctcagcggcctg2761gtcaatgaggttggatcgttggcccagaaagccacttcctgccgcgactggagccagccg2821tcgttgtgaaaattccccccacggggaaggaagtggctcttgggctgtttattttggctg2881cgggctagcagacggcgccaggggcagcggccacgcagaccgggcatttgtctagctggg2941cgggaggcggcggagcgggaagtgccacagattgaggggcgcacggcggtggagggaggg3001gcgcccagtggagagggggctcacggtgacagaggaagtggcgccgcttgggggggtgca3061cgatggcagggggaggggtgctcggtggagggaggggcacacggtggcggggggaggggc3121gtgccgcagattgagggcgaacggtggcttggggaggggcgctccttggagttgggggcg3181cacgatggcgggggcaggggtgccccttgggagcagaagcagagaagactcggaactcca3241agagcaggtggatctgggagccccagatcctgggcactgggttgggggagaaaggggtca3301aggtgtgcccgcccccgggatgctcggactctggggatccaggcctggtgacccccggtg3361aggaagagccagtgggaggaaagggccgggcgctgcccggagcagaggctggaggcgaga3421gcgattgctgcttgctgtagcgcggccgctctccagttcctccggcaaggggggcctggg3481ggaggactggccgaaaatactgtgatgctttggtctaagacaaaagccagcaggggatga3541gatcaaacaccatgacactgcgaaggggaaggccagcgcgggtgaggatttgccttccca3601gagaaccccaggggcggggctagcttccactcagcccgcattacccccacccccgccccc3661acccccggggcggggtctggaggatcagcaaaaccgcgggaaaggtgacagtcgccttcc3721ccaccaccgcctgcatttagtaggtggagggacagtcccacctctccacccattccctgc3781cagagtgtgatttaatgtcttcctagaacacaggatatcacagtctgcagcacagagatt3841tgaagtggggagaaaagcatctctggaaatgactggtgggggaagcccgccactgctcta3901caaagggtggcctgtgacagcaacagcgcaacactgggactgtgtgtgtaggttttaggc3961acaagcactttaagctgtgggaggaaagcagtttggcctggctgggatggaggggctgtt4021ggggaggagggaggagggtgtgaaaaggggaaggtggccggtagagggaaacccaccatg4081gttcaggatgtcggtggggatgttgcacagttgggcggggtattgggcttcgatatcatg4141taacccccgtcctccatggaagcaatgcccccctcgtccagcccaggatctaatgtgcat4201gcatgtacgtggacacagtctctagccaggggtggtgcagggtggggtgctgggtcccca4261gggttggcctcttagcagggtggggctggcagcttctgctgactcaaggacatttccagg4321cctggcctggaggttaagcagctcattgaaaacagctgctcctcttggctggcggccctt4381tcaggcggcctctggctctccctgggatgacccctcgcctctgcgtggcctggggagggg4441cagtgaagggagagaggctcagaggctggaggccaccctgccatgccctcagggatggaa4501ctgagtcccctccagggctggccagctgtcctccagcccccagctttccaggagctaagc4561caggtggtcgctgggccctgcgttcctgcctgcagcctttgctcctgcggttgggcagct4621tggaccgccttccttgtccttggaatgccttccttgtccttgtctctgcttgtgcctttc4681agggttttatgtgacatgcctctcattcaacacatatttgttgagagtccgctgcatgcc4741aggcaccatgctagaattaacttctcatctgcctccttcatctgtgatgtcttgtggaga4801gcagccgtcttatctgaggtcagacctctggctgcctctgtggatgggctgagtccgggg4861aaactgcagcaacagctgagggttttgctccagagacacaaactgttgtcaagcctttgc4921actcagcctcgcgacttcaccaaagcattgtgtgccttgaagcagacaggacgggagagg4981aggcctcagtgaccatcaaggaggaaggagatgggttactcaatgtagggcaaattgctt5041taaaacccagttgcccatgagcacaggcatgtttcccaggtgatgacctggatgtctgag5101aaacaagaagtgtggactacatcagaaatattcctatcaaactagacacggtggctcaca5161cctagaaatcctgcactttggaaggccaaggtaggaggatcaggaggttggggccaggag5221ttcaagatcagcttgggcaacaaagcaagactccgtctctacaaaaagtaaaaacaatta5281gctggggctgcgtgcggtggctcacacctgtaatcccagcactttgggaggccgaggcag5341gcggatcacgaggtcaggagatcgagaccatcctggctaatgcggtgaaaccctgtctct5401actataaatacaaaaaaattagccgggcgtggtggcgggcgcctgtagtcccagctactc5461gggaggctgaggcaggagaatggcatgaaccccaggggtggagcctgcagtgagtggaga5521tcgctccactgcactccaggctgggcaacagagcagactccgtctcaaacacaaacactt5581tgggaggccaaggtgggtagatcacgaggtcaggagattgagaccatcctggctaacatg5641gtgaaaccccgtttctactaaaaatacaaaaaaattagccgggcatggtggcgggcgcct5701gtagtcccagctactcgggaggctgaggcaggagaatggtgtgaaccctggaggaggagc5761ttgcagtgagccgagatcgcgccactgcactccagcctgggcaacagagcgagactccat5821ctcaaaacaaacaaacaaacaaacaacaattagctgggcatggtggggtgcatgcctata5881gtcccagttattctgggggcttaggtgggaggatcacttgagcccaggaattcaaggctg5941cagtgagctatgatcatgccactgcactccagcctggacaacagaacgaaactctgtctc6001tttaaaaaaaaaaaaaaaaagaaaaagaaaagaaaaaaagaaagaaagaaatatttctat6061cacacttcatttatctggaaagttcatttctatctatcaccttcacccccaaccgtgcag6121acttaaacttttccatggattgcttcatgagttcagcctctccagcttcattataaacgc6181atgagggcagaggctgtatcttcttttgattcctaggctggaatgcagtggcgtgattat6241agctcactactgcctcagactcctcggctcagttattcctcctgcctcagactcccgagt6301agctgggaccacaggcacatgccaccacacctggctaatttttttttgagatggaacatc6361actctgttgccaggttggagtacagtggtgcaatctcagctcactgcaacctccgcctcc6421tgggttcaagcgattctcctacctcagcctcctgagtagttgggattacaggtgtgcgcc6481accatgccccgctaatttttgtacttttagtagagatggagtttcaccatgttggccagg6541ctggtctcaaactcctgacctcgtgattcacccgcctcggcctcccaaagtgctgggatt6601acaggtgtgagccaccacgcccagcctacacctggctaatttttaaatttttttgtagag6661acgggatctcaccatgttacctaggcttgtcttgaactcctgggctcaaacgatcttccc6721accttagcctttcaaagtgcagggattacaggcatgagccactgtgcccagcatctggtt6781catttttaatttcccttacagtgcttcataaatgattatcatctgtatgggataagaaag6841cccccaagggagcagccagaaggatgattaggagaatgggctgagggtccagcctccagg6901cacctcctctgacatggccttggtgctgcagccgacccaagaggctggtctgacaggtca6961gaggaggtcctagagcagtaacctcccaggaaataaggagaatgggacagaaacccaatg7021cagacaagctaattaaaaaaggaatgcaggagcccagctggctgcagatgatcctgagtg7081tttcacagaatggaggaaagagcctcagggattcagggacaggacatccagcatctttgg7141gactcacttcctctctttccatctctttgtcccgtttcactctgcttctctttatatatt7201atcccctcatcgtaggaagcatggccattggcagccccagcctagcaatcttaacaaaaa7261gggaaagctttatgtgttccaggggagttctctgatcagccctgtttgagttatgtgccc7321catcactgtggccaaggggatagacactgtcatcagctcaagagtgagccttgtgcccac7381ccacacatccacatagccagaggtacacaatacaaggagaacaagaaggggatgggaagt7441ttcctgggcagataaaggcagctcaatgaggtcacagattgttatcgttgccatgtcttc7501attgtagagcccagcgccctgccagaccatatggtgataagggctgggatcatgtctata7561ttgttcacaataccctcagtgcctRatgccctgctggcatctatgaaattctcaatacat7621cattgttggatgaatgaatgaataaatgaatgaatcacacccagccagtgctcagtgcac7681agctccctgcctagcttcattcatttctccatttactcagtaagaacatattgagccaca7741gcttgatgcgtagcactgtgctaggcatgggttgattgcctacaaaatgcaagaagtggt7801ctttgtccttggggaggagaatcccaagatccttgtgaggggccctgcaccttctgtggg7861gggcacttttgctccctgcactgacacctgatgctcccatgcatatggatgaggctgggg7921ggtgggtgaggctggaggggccctctctggatgggggtggggggcatcaggcctctgcac7981tctgagctccctctgcccgctttgtatgtgcttgatgaagcacgSgtacaaagaacacag8041agccagctctcaggacgccttttctctttccccagcagttccagcccagcctgagcgtgc8101actccccctccactaggccttggagaaagagcctcttttcattgagggcccaggcccacc8161ggccacttcgggtggaaaatgctctcaagaaccccctgaccccggcttggcagagtcccc8221ccggatccacctacaggaatcccccctcagcttctcctttgctttgcagagctcctgcac8281gcctctcctctgctttttgattcctcagccttttctcatttgctgccaaatgagaaaatg8341caagaggttgcctttgtcctcttttgctgccaaatgagaaaatgtgacctgtgttcttca8401gtgcctcctccacattgcagtcttcagatctctgcccagcggagtctctcggcccttggg8461aggcacagagctccttgtaacgcccccagacacctgggattttaaaggtttgtttgctct8521tatgcccggcatttctcctctaactggattacaaatcctgcgaggggagggacggtgctg8581cgagtttccttgtggtcccagcattgcctggaacatgccatgcacctagcagaaagttgg8641gtggatttgaacccagagcctaggatgtcagagggaggaggggcctYgaagctgtgcctc8701atgtgacaaaggaggaaactgaggcctgaaaggtacttctgttggacattctaatcccgt8761cccctttctggacttcagtttcctctgtcaagtgaggggaggttggcctgaccctgggaa8821aagcccctcccgactaacctggacctctttcagacccttcattcaccgtttgctacaact8881gctacagcccacctggcattgatttagcacacttgtggattttctggctggggaggagag8941gaagataacgtcctcttgggctattgtgggctcacgtgaagttttgggcttcgttttcca9001gtcgtatctctgagaaacatctacaaaacaagcaataatttttttccttttccggcccca9061ggcccctttggctacctcctgccaaggccattttctgttactactgcctggagtagagtt9121tctgagaaccagatacctactccttaaaggaagcttgggcaggcatggatcaagtcctgg9181agctggtttctggtttctggtgggtggctgaaaggtgacctagtctgtttgggttgttat9241aacacaataccgttaacagatggcttgtaaacaatagaaatgtatttctcatggctttgg9301aggctgagaagtctaagatcaaggttcctgccgattcaatgtctggtgagggccacttcc9361tcgtggacagc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gtgcctcttaactggttatctgtacctgt60181tactgctcggtggtctaagtgggtgagatcaatttcagggtttctccagttcaatgtgag60241cttgtggtgagagaagagtgtccctggtccctcctgctgctttttgctctggttcccagc60301ttgctccttgcaacttcttcctagtccaggacagatttactggaagtttctttagaggca60361gtgctttagggctttggtcttcatctgttgggttgtactgtttgtcgtttgtcatgtttg60421ctttgcaagcctggtggctaatgttgcttgcattggctcactgctcttggggccatctgt60481aacctgtatcaaccccggatgctttgaagttcagtgtgcaataaaaagttcattaaaaag60541atgtggaatcatgaaaataaggttacagaaatgttaacaaaggatcatatcctgcttctg60601gtacttttcttttttaaaaaatgtatttctttgagaaatttatgtcagcatttaaaccat60661ggaagagtttaggaaggacaggcctgggtcaaaaggagggtgagattcttgaggagatat60721cttggcagagccccagcctgttgtcttaattgagagctcagggtctaaggagagaatgtg60781agccagaaaaaaaaaaatctaggtttcttatgactaacagtttgaacccagctttttttt60841ttttttttttttttttttgagacggagtctcattctgtcacccaggctggagtgcagtgg60901cacaatctcagctcattgcaacctctgtcttctgggttccagcgattctcctgcctcaac60961ctcccaagtagctgggattacaggcgtgtgccaccacacccagctaatttttttgtaatt61021ttagtaaagacagggtttcaccatgttggccaggttggcctcaaattcctgagctcaagt61081gatcctctagcctcggcctcccaaagcgctgggattacaggcatgagccatcacacccag61141ccaggttcctctttttcgcaggtcctccatgttcgggtgcccacacgtaatacacaacca61201gcccctgcatccttgctctcctctatttttggggggcaaatatcctagaggctcattcgg61261ttatatgaccattgatgaccatttaattactactcatgtttaataataagcaattatgtg61321atcgtcacttacttttcagccagagggtcaccatgtgtacacttagcagctgctaaatct61381tagtcctaacaatgatagtcccatatcctcccaagcgaccgtgaggacctcaacagcttg61441cacatctaataaacagcactgcagggttttcagggctgccaccggacagctctgggtcac61501agccagacttaccctctctctttccacctggttgtggttgagcctcatgagttctgcaaa61561cctctgctcatagaggtgaagcagcagcaccaggtacaagcctgaattcatcagctcgtt61621ttgtgcctacaaatggaaaagaaagctcttcagttcatctgtaaaatagtcatggtctaa61681attgtgggatgcagggtaacaaggctaagaggtgacaagaactgggagaagagcccaagc61741tccaaggcaaggcctgctcttggggaccagaagacagagcaggaggagatccagggtaga61801ggccagggcagccactcccctgggattttttactagctagtggaggatgctcggagaaag61861ttccattttccaggctcaatcccattccacatttattgagcagttacatatgcaaagcct61921tgagctggatgttgtggggaacacagggcaaaaagcctcagaactccctgctgggaaccg61981atcacctggtgtgggggagagagcttcatcggtaaacagctccagtacaagctggatgtg62041tcgggtgctacaacagagatgcaccaggccttgtccataacgttctcagcagtcacagat62101gtccccttgtgaaaggcttcctgaggagcctcctttgcccccatcattcctttgcttccc62161agtccctaagcattcgtcttgatcttgcatatcaccttttgcacttgttaacgtttccgc62221tttctcttgaggagtgggctgtgctttttttctcgctggtgctcagaacaggcgctacac62281agggaggatgtgatggtctggctgcccatggcacttctctgtccttagcacttgggcttt62341gctgccaagagtggctgtgctcactctagacctttcacagaccttccataaatcagtggt62401attagtaacaactactcctatccctctataggggaaggaggggaaggccctttcagtccc62461catctttcccttgagtgggggtcttgggatgggccaggctgtgctcacaaagccacccag62521cccccggggaatagagtccccagcagagggcatggccccttccgtcacccttgggaaagc62581ttgcaactgtgactctgtcggcccaggataacgatgcgtgggcatgaatgacacagtgag62641cactcagtgtctcatctgccttcttcctgcagcagcgcgggtgccacatcttcctgttct62701cattagatccagggcagctcaggaggttggcacagaacccacctctcccaggaccaagca62761gctgccagaaaaggcacagaagttgagcaaggattgatatgctcttccttctgaatcagg62821gccaactctggatcttggtgacagccggccctgagatcactgctgcaggcctctccacca62881ttctacagatccatcttcgaaggccccagtcctctgtccttagcagttagcctcctattt62941ttagcctttggtcgtgttttttgtgctctgctgaccttggcctagagtgagtagagaaga63001agttgcttgtttgcattatttctgttttatttatttaattttatttttgagacagacaca63061gtcttgcccaggctggagtgcaatggcgtgatctcggctcactgcaacctccacctccca63121ggttcaagcaattctcctgcctcagcctcctgagtagctgggattacaggtgtgcaccac63181cacgcccagctaatttttgtatttttagtagagatggggtttcatcatgttggccaggct63241ggtctcgaactcctggcctcaggtgatccacctgccttggcctcccaaagtgctgggatt63301acaggtgtaagccaccctgcctagccactttttctgttttaattgcctcctcctgccctc63361cccgtgagtctgtgcctgcctccgactgtctcgccaaccacctcttctgagaagccttcc63421tcacctggcttttagctcctcctcaccaacaaagccttggtccttatttgtttacctcat63481ccccggcttcctcctgcaaaagctgccagtggttctctcaccagtgctctttgccagcca63541gcttttatgcttgagacaccgggatgcatgatgacaaatcaatcagctagaaaatggttt63601ggaactatcaggtttgtcaaagcattcaggaaaatcatttactttttatgactcaataag63661gcccagaggcatgcagatgggagaatatccactaatgttttctgcagataatttgaaaag63721tcacttctagcagattgtgtctcttctctgatctgtgggagggaacctgccacacagtgc63781catcaatctcgccggctcggttcctttatgtgcgccctttgcagagctggtgatgctcga63841caaaggggctcagcccaccaatcagagcaggcgtgtgatagtcaactccgaatcagcctt63901tacactcctgagctacaagtgggagggatttgttaactcaaagagttccaccgcacttgg63961ctggccagggccaaatgaacgaggttttggagcagggcttactggagctacggcccatcc64021accccttccttgtcccaatcacatcacactttgactcccacgtggagttccctttgtgtg64081ccagactgaagggaatgggggtggcattaataggagggggcggggacattaaaaaggaga64141aggcatgtgaaacagcgcaaaagggctccggagatctcagcccacctgccagctgttttc64201tccattttgctaatattcatttaggccaagagacttgctactgacggaaattctcaatat64261attccacttaaagaagctgacagcctggctgaatgttctatcaagtcttcaaatatattc64321aagagtgctcctaggtataagggtgattaggtgactgcgtgtgtctgaggacatctgtcc64381ttggctgagatggagccaggccaggcctgtgcttcacagcaatgacagcctgaggcccac64441tgagactctcctgtgcggcagagcaaggatggcaggggtgtgttccaattcagtgagcag64501gggcccctggagtgcatcttacctggaaagcgctgccggctaaggtgggcagtgcagggc64561caccggacactgtgactttaggcagagttttaactgtgcaggaccaaagggactcaattt64621gtaatggaagggaagtaacctctgttctgcctgtttcctcaaagcatgatagtaaagtga64681gctaatatatgggatctcgcaacaccaacacatcctacaagagacaagatttaaagacag64741gattccaggtaaattgtctgggcgttaagatctgtgagggcaagacctgcatctgtcttg64801tttgctcctgactcctagtgcctggcccagtgcctggtatactgtgcatgctttataaat64861atatgcacagatgaagcaatgagaaaaaaatgcataattgtgatggatcagacagtgacc64921cattcagccacaacttttgaatctagtggctccaggtttaacatgcatcagaaatactga64981gcaggccttttcaaacacagttgccaggcagtcccgtcccctcccctcccccttctcctc65041tttttttgagacagtctcactctgttgcccaggctggagtgcaatggtgtgatctcggct65101cactgcaacctccgcctcccaggtttaagtgattctcctgcctcagcctcccgagtagct65161gggatttcaggtgtgtgccaccactcccagctaatttttgtatttttagtagagacgggg65221tttcaccacatttggccaggctggtctccaactcctgacctcaaatgatccacccacctc65281ggcctttcaaagtgttgcgattacaggcgtgagtcaccgcgcctggcctcactgctcctc65341ctttcctccctgcatttctgattcagctggactggagtgggcccaagaatgtgtacttct65401aacaagttcccagatgatgctaacgctcctggtttggggaccacccatttttgagaacca65461ctgatctaaactaagggtgagcacgtggtagctgctaagctgaatctggcctgtggatgt65521gctctggttggatggaaggaagcaagaaggaaggtagaaaagaagagcatgaccatcttt65581agttggggagagcactttccattttgccacaaatgccaccacgcccaatggcaggatacc65641tagaagattaccatcatttgttgtctgcctggcctctgtggtatttgagtgtgtaacctg65701tgacctaagctttccttaaaYctgttttcacctcatccattttggaggctagaatgttat65761ttggctcgccgcatgctgctgtccaggagttctgccctccaccgcaaaaccgagtgccca65821accagcatccaatatgagtcctgcttccaggcgacagccaacgctcacaggagcaaatgt65881aggcttgcctcacacacagcactaccgaaagagtcaggagaaaggagggctgtactgcca65941caggccctgacagacctaagatgaaaccgccaggtccctggggtcccacagctgctcttg66001tctaatgatcccctctccagatgcttgtaggagttcctgatctagccgcttgaggcctgg66061taacagaccttgaccttgggcctcatactagaataatccgagttcagtgccctgttagca66121cctgtgagcagcatctgccctgctcccaccagaagtacagaactaaggtaggaaaacgtg66181ctctgacgggagcccctccacactctcaaaggccaccgacagtgctgacccggcacctag66241aggacacgccctgatctgtgctcaggtcccaggggaggcgtccgtcccagatgtcacaga66301cttcactccaaggcccggttgatagaaggcccgctcctctctttccttccgctgttaaac66361atctcttcttcagaaaggccttctctggttcagagagcaagtcggagtcccacagctgtt66421cccgcagcggcctgcatggtccttgttttgacctccacctgcaggaattgctggcttcag66481tcttcgctgctggtcggtagggccctgcgggtcttgggtacggcagcattttcggcacag66541tatctcacaggcagcaggtgctcaataagtatttgaaaatttagttaaactggctgggtg66601tggtagctcacacctgtaaccctagcactttgggaggccaaggtgggaagatcacttgag66661cccaagattttgagaccagcctgggcaacaaagtgagacagaccccatctctcaaaaaaa66721aaaaaaaagaaaaaaagcagggtgtggtggcacacacctgtggacccagctacttaggag66781gctgaggcaggaggatcccttgaacccaggagggaccctggccccatcccagagctgggg66841ttagacctgggttgggcctgtgtatcaccattttaatagaaaaaaagaagacctgcagct66901aaatgttataagttatcattttgggtggtaaattataagaaatttttgtatggtttttgt66961aattaaacaaaagcaaaaaaaaaaaaattctaagtgggaaaatatataagaaaaacagtt67021cactagtcagttagtttgaggttaagtgagaaccaggctaagctgaagcccgttctaggt67081ttcttttggttttgccccttttttggtacatgtctagacttgtacacgtctagaagtggt67141gccacttctccagaaaggaacccatttgccaaggacaagcagcaggagggtggccaggag67201gcccaggtgagccaggtttggaagcgaggggaagggaccaggcccacaaggcacaggtgt67261ccatgtcacctccacggccaggctggtggcctcctcagctgggcttgtcccctcagtggg67321ctcaccgcgcctcggggacgagctgagtttcagtgacgttgtagatatctttgtgttcaa67381caacagttgcttcagtctttatctatggcgagcttcagaattcgcccaagtcaggctccc67441tggggcctggggtttcgcccaggctgtgagcagacaggcactatgaattcccagcaggtc67501atggggacagtcagggctgcagtgaggtgtgagccctcagccgcctccagacagcagccg67561cccatgctatgacctggctctccccacagggcgggcctggcagcctagctgtcacctggg67621atgataatgcgggcatttcccaaaggctctgatggcttgtgaagttctcactgaaacgaa67681tttaaaatcattcctgcttccacgtggagcaatgactgtgctggatgctttctgtctctc61741ctgcagatccgcacccctcctgctctgtggcccaggggccggccccatggctttccccag67801ccgggcttccttgccctctggcttcccgttgggtttggcccatgggaggccccggcagga67861gacccgagggcggaggagggagaagctgggtgtctcttcccaatccctctctgctgggcg67921tggtgtgacagtggttgctgtcttctcctgaaggccacaggctgctggtggctgatcctt67981agctacagcgctcaKgaccctggccacactgctagtcccaggacgcctcctcgttccttg68041ctgctttcttttgctgcccagatttccctcctcccttctgcgttcttcacttgccccctt68101tgagtgtgccaccatctcttcccacccagatcctggctgatacaattactgtcatagttc68161tttctagaaggtgcttgtagccagttgtcttaacatggag


Following is a CDK10 genomic nucleotide sequence (SEQ ID NO: 4).

>16:89456451-895557001gttaggaagg gtgcagtatt ctaaagtgta ctgcagacgt gtgtaaacgc catgcctgac61acacactcaa gcccccacgc cagccggagc aggacggggc agcagcaggg cggctcgctt121gggggcggct gcagctgggg cctctgagtc gagatcaggg ctccctggtc ggagcctcag181cccaagctct cctccagcct ctaacccaca accctctctc ccttcccaca gcaYcagagc241gtggggttgg tgacacagcg tttcggggac cgaccctacc tccgtgacag ctggtcctcc301tggctgcgca cagagctctc gcccacggca gaggcgccct cgggcagctg gctgagctgg361tccagcacct ccaggccatt ctcctcggcg atctgcatga tgaggctgtc cacctgctcc421tgcggcgtgg tcagggtggt ggccgagctc atggagtcct ccatcacctg ggggcagggg481catgctctgg acaacaggtg gggccaggcc catggggcYc cactcagctt cacaagggta541cgactgcact tttcgttccc tcacacagcc agccaccttc ttgccctgtt ctctcagtga601ctctctctgt ctcaccttcc accaggagcc tttcctgccc ccccacccag ccccaccctc661tgactgtgcc atgcctgctg ccgcgggtgg acatctttcc acacctctgc atgcttgtct721gccatccgag cctccccaca aggagccagg tgcctgcagg aggcccccgc cgccctgccc781accctacctg tcagggtcct ggcagggacc cagcacagag gatgcttggt gacgtcaggg841agcaaccaca ggtatgcaga gacagcacgt cagcctgtga gccacccctc ccgatgagct901gcccacacac gctccttgtc cccaggccac agccccaagg gtaggggcca cataccgatg961tatggacgtc caggttctgc acctgctgct cgaacctgtc catcactgag gagaccttct1021gcaggtccat ggtgctcagg gccttgtcca gggctttggt cacctgggcc atattcttgg1081tcacctgaga caggagagag cgcaggaggg aacaggatga aaggcaagtg gagctcacgc1141accagggacg ggtcccctgg actctgacag gagagctggg gcaccccaac cctgacccac1201aaaaccccaa ctctgggctg agccatcatc tgggtcctat ggaaagacca ccaactcgac1261ggaaaaggcc atggagaccc agtgcggggt cagtggagaa aaggccgccg acgtggagac1321ccagtgcggg gtcagtggag aaaaggccgc cgacgtggag acccagtgcg gggtcggtgg1381agaaaaggcc gccgacgtgg ggacccagcg cggggtcggt ggagaaaagg ccgccgacgt1441ggggacccag cgcggggtcg gtggagaaaa ggccgccgac gtggggaccc agcgcggggt1501cggtggagaa aaggccgccg acgtggagac ccagcgcggg gtcggtggag aaaaggccgc1561cgacgtgggg acccagcgcg gggtcggtgg agaaaaggcc gccgacgtgg agacccagcg1621cggggtccgt ggagaaaagg ccgccgacgt ggacacccag cgcggggtcg gtggagaaaa1681ggccgccgac gtggggaccc agcgcggggt cggtggagaa aaggccgccg acgtggggac1741ccagcgcggg gtcggtggag aaaaggccgc cgacgtgggg acccagcgcg gggtcggtgg1801agaaaaggcc gccgacgtgg ggacccagcg cggggtcggt ggagaaaagg ccgccgacgt1861ggggacccag cgcggggtcg gtggagaaaa ggccgccgac gtggagaccc agcgcggggt1921cggtggagaa aaggccgccg acgtggagac ccagcgcggg gtcggtggag aaaaggccgc1981cgacgtggag acccagcgcg gggtcggtgg agaaaaggcc gccgacgtgg agacccagcg2041cggggtcggt ggagaaaagg ccgccgacgt ggagacccag cgcggggtcg gtggagaaaa2101ggccgccgac gtggagaccc agcgcggggt cggtggagaa aaggccgccg acgtggggac2161ccagcgcggg gtcggtggag aaaaggccgc cgacgtgggg tctccaggac tgctgtgccc2221tcctggccca cctgcactgt gtgaaggaaa tgcctgcaac agccaaggct ctgctgtgac2281gtccctgacc cagcagatga gcctggccat gcaggctctg cctcttgtgg gagttgccaa2341gaccgaggcc tggccaggta cagtgacagt gagtggttca cgcctataat tccagcactt2401tgggaggccg aggtgggaag attgcttgag gccaggagtt tgagaccaga gcggccaaca2461tagacacect gtctctacat aaaaatgaaa aaaatcagct gggcatggtg gtggatgcct2521gtggtctcag ctacttggga ggctgaggca ggaggatcgc ttgagcccag gaggttgaga2581ccagagtggc caacatagac accctgtctc tacttaaaaa tgaaaagatc agccgggcat2641ggtggtggat gcctgtggtc tcagctactt gcgaggctga ggcaggagga ttgcttgagc2701ccaggaggtt gaggctgcag tgagctgtga ctgcaccact gcactccagc ctgggtgaca2761gagcaagaca gtctctaaaa aatatatatt taaaaaaaaa aaagcccaag tcttatcaag2821aggtccttcc tggggcatca gtgcaagctc agggtgcagc ctcacctgtg tccctgccag2881catttcccac cccacgctga tccagcttaa ctcaacctcc ccaccccacg ctgatccagc2941cctcaacctc cccaccccac gctgatccag cttccctcaa cctccccacc ccRcgctgat3001ccaccttccg tcaactctga gctgccccag atccagaccg cagagcccat tcctgcttca3061gccagcgaac gccacccatc cagcaccagg gcccagcact cacccccttc atagtcacag3121ctgtctgcac cttggaggcc actgcgtcta cgcgggacgc catccgaagc cagttcacac3181cttcgttctt cttgcggatg gcgttctcgg catacacacg ggcacactct acatttttct3241gcagaagggc ctgaaacccg Sgggggaaag cagctggaag agcttggtgg tgtgtgtgcc3301ccctgctagg agcccagttt ttgttttgtt ttgttttgtt ttgtttgaga cggagtcttg3361ctctgttgcc caggctggag tgcagtggca cgatctcgac tcaacgcaag ctccgccccc3421caggttcatg ccattctcct gcctcagcct cccgagtagt tgggactaca ggcgcccgcc3481accatgcctg gctaattttt ttgtattttt agtagagatg gggtttcacc gtgttagcca3541ggatggcctc gatctcctga cctcgtgatc tgcccgcctc ggcctcccaa agagcYggga3601tcacaggcgt gagccaccgM gcccggccct aggagcccag tgttgaggcg gtgggcgggc3661ctctccccag aaatgtttga gctaaaaatg caaacaccca ggccccagcc cagcccccca3721agcaggcttc tgcttccccc cgcacccctc cagcactcca ctgggcacgg gacacagcgg3781gtgacactca gagcctgtgg caggcagttc tccgagctcc attctccctg ggagcctggc3841tgtgggtgaa gagttaccaa agtccaatac gctgggacct acttcaccga ttacgcagtt3901ttaagagctg cacaagatcc taccttcttt tctttttttt ctttttttgg agatggaatc3961tcgctctgtg gcccaggatg gagtgcagca gtgtgatctc ggctcactgc aacctccacc4021tcctgggctc aagcaattct cccgcctcag ccttccgagt agctgggatt ataggtgtga4081atcactgcgc ccggcctaga tccctcctct ccttctgccc tgtccatcat ggggagggga4141cccctcctgg agaggcaccc ctgtaagcct ccagccccat cccaccccat ggggcagggg4201catcatgact ctgcccagac cctgatgccc tagacaccag ctctgtaggg aggagagags4261taaaaatgat aggatccgct gggcccagtg gctcatgcct gtaatcccag cactctggga4321ggctgaggca ggtggatcac ctgaggtcag gagtttgaca ccagcctggc caacacggcg4381aaaccccatc tctactaaaa atacaaaaat tagccaggtg tggtggcgca tgcttgtaat4441cccagctacc caggaggttg aggcaggaga atcactcgaa cccaggaggc agaggttgca4501gtgagccgag accgcacctc tgcactccag cctgggcaac agcgagactc catctcaaaa4561aaaaaaaaga taggatcgtg gcagacagct agctagcagc cctcctcctg cagatggcaa4621aatcaagttc agaatgttgg aaatgcaaag aaggaaaaca atgaaggcaa tgcactcatc4681tctgtgctct ggacgccctc agtgggctgg gaaggtctgg caggggctgt agggcaggga4741cRgaaaccat tctgggctgc accacacaca gcacctccct tcctgagtgc ctgtttgtct4801tatgtgggaa gcaaatactt tgaggctcag aaatcatagt gaggattagg ctgggcacag4861tggctcatgc ctgcaatacc agcacttttg gaggccaagg caggtgggtc acttgaggat4921aagagttcga gaccggcctg gccaacatgg cgaaacccca tctctacaaa aaaattaaaa4981aattagctgg gtgcaagcct gtaatcgcag ctacgtgtga ggcgaaggcc ctagaatcac5041ttgaacccgg gaggcggagg ttgcagtgag acgagatcgc acctgtgcac tccagcctgg5101gcgacagagc aagactccgt ctcaaaaaac aaacaaacaa acaaacaaac aaacaaaata5161tatatatata tgtacctcaa ggtgacgatt aaatcatgat gtagaaagtg ccctgcccct5221ccccaaaagg catcaaaaca aaacaggaca caaaaataag ggcagaaaag gacagaagac5281aaaccaggag agtcatgacc ccacagcccc cagggtctca ccttcttcac tttggcctgc5341tccgccttgg agtccttctc cgccttcttg gccagcttct ccagctgctt cgccgtgaac5401tgagcggaag ccggaatgtc ctgggtcaga catgcggagc ccatccccca ggcccggctc5461tccacacccc cacacctgtg cccacacctg tgcccacaca ggttcctaag cccccatcag5521ccagcctggg cccagcacac cgcagggaag gcgtgaagcc tgtgggcgtg catcctgggc5581ggcgagagat ggcaccagct ggaggaggaa gctgcctgct ctgcctcagg ctctgtctgg5641gttttctcca gtgtctgtct tctctgtgaa cagcctcacc caaggcctcc tccagagcgg5701aagggtcaaa aacctctcac tctgatctcg gctcctgctt atggatggct tttcaacgtc5761aaagaaagac tgaaggaaac tcatttcagg ctacccccaa gtcataacac agaactccag5821gtaaaaacac acctgaaagt gttatctcga gagaatgctg tggatgcagg aggttgctgg5881gcagcacaag cagcttccgg ggatctaact gctctgccag caccacacgg aaacagggcc5941tctggcaaca ccacaaggac acaggacctc tggcaaccca cccggaacca cagacgccct6001ggtaccaggt agccctggcc accagcacct caaggaaaca gggcctctgg caacccatct6061ggaatcacag acaccctggt atcaggtagc cctggccacc agcacctcaa ggaaacaggg6121cctctggcaa cccatctgga accacagacg ccctggtacc aggtagccct ggccaccagc6181acctcgagga aacagggcct ctggcaaccc atctggaacc acagacgccc tggtaccagg6241tggccctggc caccagcacc tcgaggaaac agggcctctg gcaacccatc tggaaccaca6301gacgccctgg taccaggtgg ccctggccac cagcacctcg aggaaacagg gcctctggca6361acccatctgg aaccacagac gccctggtac caggtagccc tggccaccac agcaccacag6421ggtgcaggac acaagcaggc ctcagccggt cccctctgtg cccaacagca actgacaggc6481ccaccgtgat gggaaaacat cccaaagctg catacggtag ctccggcaca tcactcctgc6541ccagtagctc agagccagcg acaaggcaag ataaagctgg ccacagatga gcagccagga6601gcacatccca gggaaatgca cggaaacctg ccctggatta ccggaagctg caacctccac6661cccacgtggg aggaatactc agaaagctct ccttcaaatg atctcaaaga atgagtccac6721acagcggaac accaaggcct aatgcccaag ttgtcttttt tttttttctt ttcttttttt6781tttttttttt ttgagacgga gtctctgtca cccaggctgg agtgcagtgg cacaatcttg6841gcttactgca agctctgcct cccaggttca cgccattctc cggcctcagc ctcctgagta6901gctgggacta caggtgcctg ccaccgcgcc cggctaattt ttttgtgtgt ttttagtaga6961gatggggttt cactgtggtc tccatctcct gaccttgtga tccaccccac ctcggtctcc7021caaagtgctg ggattacagg cgtgagccac cgcgcctggc cccaagttgt ctttcttaag7081ataacactcc gggccaggca cagtggctca cgcctgtaat cccagcactt taggaggccg7141aggtaggcgg atcacgaggt gaagagatcg agaccatcct ggccaacgtg gtgaaacccc7201gtctctatta aaaatgtaaa aaaaaattag ctgggcatgg tggtgggcgc ctgtagtccc7261agctactcag gaggctaagg caggagaatc gcttgaaccc gggaggcaga ggttgcagtg7321agccgagatc acgccaatgc actccagcct gggcgacgga gtgagacgcc gtctcaaaaa7381aaaaaaaaaa aaaaaaaaag ataacactat ttctttcatg gctttcatgg atgaccctaa7441ggatctgcct ccacttagcc tcagataaga aaagagctaa tatgtcacca aggtggctca7501ataatcccct ccttggtcag cagtctgagc cctgtgtggc ggtcactcaa ctgttatata7561atctgcccga ctgtttctgt ggctctgagt gagcgtggct tatactatct gtcctcacca7621gaacagggct ggggtgaacg gccattcggt acataccttc aactggaaca gggtatctgc7681aaagaaagag ggaattaatg gtttgaggat tgggaatggg acgttacact tctggcaggc7741aggactcact aggttccttc tagacaccag gagctagaac acacacacag accacacctg7801cctggggcct tcggggagcc ccccccaaca ggggaggcac gagacagatt cgtgtgccag7861ccctggctcc ttgctagcca cagggcccag caaatgcaga ccctccctcc ctgtttcctt7921gttgggaaca tgagggtggt accagccaat ctcaaagggt tagtgtgagg acagaagagg7981tcatggacgc agcatctttt ggccatggtt agaatccaaa agatggtggt ttataattaa8041tgcaactttg caaaWgtccc ttgggattct ttcccaattt aaaagttcac ctgtaatccc8101agcactctgg gggtgccgag gccagtggat cacttaaagc cagggtttga ggccagcctg8161ggccaattta aaagtKcagt acttgacagt ttacttttta cagactcttt ctcctcaaag8221gtaaatggat aaaaatagca aggtggtaac tcttagacat caaagagctg atttccagcg8281gtacagtcRc tcacgcctgt aatcacagca ctctgggagg ctgaggcggg cagatcactt8341gaggtcagga gttccagacc agcctaggga atatagtcag acctcatctc tattagaaaa8401atatatattt ttttgccggg tgcggtggct cacgcctgta atcccagcac tttgggaggc8461cgaggccggc ggaacatgag gtcaggagat cgagaccatc ctggctaaca cggtaaaagc8521ccgtctctac caaaaaatac aaaaaattag ccaggcgtgg tggcgggcgc ctgtagtccc8581agctactcag gaggctgagg caggagaatg gcgtgaaccc gggaggcaga gcttgcagtg8641agccgagatt gcgccactgc actccagcct gggcaacagt gcaagactcc gtctcaaaaa8701aaaaaagaaa aaatattttt tttaaaaagt aggattgatc tcaattcatt caactgctaa8761atttggagag cgaccctgac cccagattcc tgaaagggct ttaccacact agccacaacg8821aggaaatgga gcctgtaaaa gaatgaggtt actgagtgtt cacaaatgca caccaaggct8881ccaagatgag atctcccggg gttccagaaa gctctgggca tcgttcaatc tatcgcctgt8941cagtgcaggc gtggtcaagg ccaagttcaa cctgtcatct atcagttcag gtgtggtcac9001ggccaaggtt atctgtgact ggaaaacagc cagcctgacc cagatggcac agtgcaggcc9061aggcgggccg tcccacccaa ctcactgtgg gatgctggcc aaaccgccct cctcatctca9121gctttccctc acctcagctt tccctcacct cagctttccc tcacctcagc tttccctcac9181ctcagctctc ctcacctcag ctttcctcac ctcagctttc cctcacctca gctttcctca9241cctcagcttt ctcttccagc cgtgtgtggc agcacaaaaa cccagctaag gcaaaatctg9301actctgagga aatctgacat gggcttgaga gaagtcccct caaaagagtt gagtgagctg9361tggagtgaaa agccacaatt tttctttttt ttttttgaga cggagtctca ctctatcacc9421caggctagag tgcaacggca ccatcttggc.tcactgcaac ctctgcctcc caggtttaag9481cgattctcct gtctcagcct cccaagtagt tagaattaca cgtgcacgcc accaggccca9541gctaattttt gtatttttag aagagacggg gtttcaccat gttggccagg ctggtctcaa9601actcctgacc tcaggtgatc aaccaacctc ggcctcccaa agtgttgggg ttacaggcgt9661gagccaccac atccggcctc ccaaagtgct ggaattacag ccgtgagcca ccgcgcccgg9721cctcccagag tgctRgggtt acaggcctga gccaccacac ccggcctggc cagcatattt9781tttgtggcgg gaggtagatg ttatctttca tcttctctca gggcagaaaa tctctacaca9841tttccttcat gatgatgaca cggtggtact atagcttttc acacttctaa ctaaaagaat9901tttttttgag acggagtctc gctctgtcgc ccaggctgga gtgaagtggc gcgatctcgg9961ctcactgcaa gctccgcctc ccgggttcac gccattctcc tgcctcagcc tcccgagtag10021ctgggactac aggcacccgc caccacgcct ggctaatatt tttgtatttc tagtagagat10081ggggtttcac tgtgttagcc aggatggtct cgatctcctg accttgcgat ccgccggcct10141cggcctccca aagtgctggg attacaggcg tgagccaccg cgcccagccc ttcaaaaagt10201atttttaaaa gcctgcccat cacaccagcc caacgatgtt tcagcaaaca taggacttca10261ttttgccaga ttcttcctgt tctttggtat ccctgagcac acagccggct agccccagat10321gtagaaggta agaattaggg cgacccattg agcacagaaa cgcttgtgct ggctcctgct10381caaacctctg cttccccaca tggagaatgg acgcaaagag ctgctgtgac cagggcctgg10441cactcaggga gcgctttctg tctgagtcct tctgatttgc agtgatgctg tttggggcag10501ggggtgcaac tgtcactttt tctggtgccc tctgacccga cattccaagt cccgaaaggc10561cgaacaggca tcgcacagtc ccccctaccg agctggcggg aaagggcgcc tgacgctgag10621gagtgccagg ggcgggggaa agggtcctgg gtcggtggtc gggggtccag gccctgggca10681aggggtcctg ggtcgggagt ctaggtcccg ggtccatggt tgggggtcag aggtcgaggc10741cccggggaag gcataccggg ttgagatttg gggatggagg ccccggggaa ggggtcccga10801atcgggggtc ggggatcgag gcctgggggg aggggtcccg ggtcgagggt cgaggcccgg10861ggtaaaggat ctcgggttgg gggtcggggg cctgggaagg gtcccggtcg gtggtcgggg10921gtcgaggccc gggagaaggg gtcccgggtc ggtggtagag gccctgggga aggggttccg10981ggtcggggat cgagggtcga ggcccgggaa aaggggtccc gggttggtgg tctaggccct11041ggggaagggg ttccgggtcg ggtatcgggg gacgaggctc gggaaaaggg gtcccaggtc11101ggtggtcgag gccctgggga tggggtccgg gtcggggatc gggggacgac gaccgggaaa11161aggggtccca ggtcggtggt cgaggccctg gggatggggt cccgggggat cgacggtcga11221ggcccgagct ctcctgagtc ctgcccgcgg ccccaggtgg gcggagccca cgccagtggg11281agaagcggcc ccgccccgcg cgcgagtccc cggaggacgg ccgcgacctc ttaccgtcca11341tggccacaat gacaggagca gcactcggag agggagaagg gacgccaact ccgggcggtg11401tcaggtcccg gcggcgatcg aaccgaccaa gctgcacccg gcggggactt ccggggtcgc11461cgccccactt ccggtcgcac gggatgctct cgcgagagga tggtggccgg tcccgccaga11521ggggcggggc gcggctgcgc ggcgcgcggt cgccatggtg acgcacgccg ctggcgcgag11581gaccttttgt gagtcgctcc cggctccgcg gccgcggagg tgtggggacc cgggcttgcg11641tcggaggggg cggtgggctc acccatgggc ctttccaaaa gcaaagagaa acccaggaaa11701ggtaaaggag gcgcaggcgc tgggctcccc gcgtctccgc ccctgggcgc gggcccaggg11761cggggctggg ctgggcgggg cggtgtgagc gcaggcgcca ggctcggccc ggtgcgaacc11821gttcctgccg ccgagtccgc cacggacggc gcgtttcccg cctgaggcgg ttacggaaac11881gcggagactc gagacttgaa gccagcttat tctggatcca cgcaggcgac tgaagacgat11941gggacccact gccctgcatt cggtaagtcc cgggtctaac ggatgatcca tatatttcag12001gtgaggagca aaagaaggga tccacctatt cagttccaaa atctaaggag aagttgatgg12061agaagcattc ccaggaagcc aggcaggcag acagggagtc ggagaagcct gtggacagcc12121tccacccggg ggccgggaca gccaagcacc cgccgccggc agcttcgctg gaaggtagga12181gacggcggga gggagcgaag cgaggtcagt ggcttggagg atctggggct ggggtgagga12241gcctactgta agaccctacc ggatggacac tagtagcagg cacgcgccgt aaagatgaaa12301ctggtttgtt ttgggaactt tatgtaggag gtaaatatcc agaaatctta gttgaaaaca12361taagtttaat gaaaatgtag gtcagttccc gagtgataag tgcgtaccag gtggtcgagg12421ggctggggca gtgtgcgtgt gtctcacctg aggttctcct cgagggctgg gtatcttggt12481gccactgaga atccctgagc ctccgcacac tggaacagag cattgggagg ccgtcccagg12541ctggactgta gctctccgag ggctgtgggg ataagtacat tgcggagggt gaccgaggag12601tagaacctgg gtgcgacaaa ggtgtctttt ataacagcct agtgtggcca gacctttggg12661tgtgtgtggg gaagatgagg ccccaaaagt aggaagaagc tagaatacaa agttttccat12721cagttttacc acgtggagct ggaagcgtta aatattttaa acaacggagg tgctgcctgg12781ggcggtagtt cacacctgtc ctcccagtta tttggccgag gcaggaggat cttttgagcc12841caggagttca agaccagctt gggcaacata gtgagaccct gtctctacaa aaaataaata12901tttaaaaata aaaaacaagg gcggcatgat gaggtctgtg cctaccaaca ggggaagcca12961gttcgcaggg gaggtggccc ggtcagagac ccattgctgt ggcccagtgg gagccagcaa13021ggccttgcag taggaattgc agaagggtca ggcagggaga acccgcagag cctgccgtcc13081gaggagattg ggggcctgtg ccaggaggag gtgttttaag tttcatcaca agaatgaacc13141actcggctgg gcgcagtggc tcacgcctgt aatcccagca ctttgggagg ccgaggcgtg13201tggatcactt aaggtcagga gttcaagacc agcctggcca acgtggtgaa accctgtctc13261tactaaaaat acaaaaatta gctgggcgtg gtggcatgtg cctgtaatcc cagctactca13321ggaggctgag gcaggagaat tgctcgaacc ttgaaccggg acccgggagg cagaagtagY13381ggtgagcgag atcgcaccac tgcactccag cctgggctac agagtgagac tctgtctcag13441aaaaaaaaaa aaataataag gagagaacca ctcgggtcag acaagaaaga atgtcagggg13501gatgactggc ctgctaggca gaggagctgg gcctggactg cttcacactg acacgcttgt13561aggatcaagg gctgaaagta ggaggctctg ggccttggac ttggaagaaa gacttggatt13621tgaattccca actttggaac ctcccactgg agggtggaac ccctcacagg agggcgcaac13681cccccaccgg aggatggaac cctgcaggaa tagcatcacc cttgggtggc agtttggaaa13741atgctacctg tctgtcacgg tgcttttagc tgaaagttac agaaaagctc actaaacgca13801gctttaacac agggatttaa tgtctcacga atgaagggtc agaggtggag cttccaggtt13861ggcttgttag cacaacagtg acatcaaaga ctttgcgtgt ctcttcttga ccactctcag13921tgtccacagc ccccgatgag tgtacagccc cccacacgca ggcaggactg catctactgc13981aaaggcattg ctccttgctt catattagca aggaaacctt tcccaggaag gcttcgcctg14041cccagcctct gcaggactca ttcaatggtg caaattcacc aggggctgga gggagctgtg14101aagcatggga gtgggcagga cctctgccag taacggaagt gggggaagga ggaccgcctg14161ttggaagggc cccagcagtg tctgtcacac cagagggtgg gggaggaagg tggacatgaa14221gggaaaacag cagagcaaga gtgagaacag agtttcagca gataaaatga agtgtgcacg14281ctctccatgc agtgacgggc tgagcgacag tgaggagcag gctgctgact cccttgccac14341accgtgaagg agtcccaaag gtgggacact gagtgagaca agccagagga aagaccactc14401atgttgcagt tccctttcca tggaatgtcc agaaaaggca catttacaga gagaagcaga14461tgagtggttg ctgggcagaa atgggagccg gccacagctg agcacaagag gactttggag14521ggtgatggaa atggttttcg acctgaatct gaacatggga agcactgcag tttagtagtc14581ctggtcccta agcccttcca gcccaggagc cagacctgtg agcaaacaag cctttagtga14641ttccaggctc tggctggaac cttgagtctt ctcagcttgg ggcatgcacc tcagggggag14701ccagcatcag tgtccagccc caagagcttc cctgtacgtc tcagtgagtc ttcacatgcc14761tccaactgcc tggacaacca cacgtgatac ctgtcctgcc aaacgtgtcc tgaacccata14821aaatccagag aaaagaaaat cgttttaaac tgctgaggtt tggggtaagt tactatgaag14881cagtagtgag aagaacagaa gggccatgat ggggagaaag tttggccata agttaatcaY14941tgtggcagct gttaggatat tgatgtggct tggctgtgtc ctcacccaaa tctcattttg15001aattcccaca tgtgggagga actgggtRgg aggtgattga attatggggg caggtctttc15061ctgtgctgtt ttcgtgatag tgaatgagtg tcacgagatc cgatggtttt aaaaacggga15121gtttcctgca taagctctct ctctgcctgc tgccatccat gtaagatgtg actcgctcct15181ccctaccttc cgccgtgatt gtgaggcgtc cccagccatg tggaactgta agtccattaa15241accttcttcc tgtgtaaatt attcagtctc tggtatgtct ttattagcag catgaaaacg15301gactaataca gatagatgta ttatactgtt acccctacta cttctgtata tgcttaaatt15361tttgcattac aaaattcaca aacatacaaa cattatagag aactaacatc acagatttac15421agtcacctta ttccacaatg ttggaaattc taccacaaat aaagaacttg aaaaaaaagg15481aggggatgtt agaaggaaat ctgagcgacc tagacctggg ctcagtcacg cctgagacag15541tcagatgggg tatctgagga tgagtcagtg ttaattttct cagacctggg ctccgtcact15601cctgagacag tcagattggg tatttgagga tgagtccgtg ttaattttct tagatgtggg15661aatgctatta tagttacgct aaaacacaat ccctatctaa taaatacact gcaacattta15721tgggtaaaat catatgtctg aaattcctta aagatactca gaatgccagg catggtggct15781catgctgtca tcccagcact ttgggaggcc gagacgggtg gatcacttga ggtcaggagt15841tcgagaccag catggccaac atggtgaaac cctgtctcta caaaaaatac aaaaaaatta15901gctgggtgtg gtggtgcaca cctgtaattc caactacttt ggaggctgaa gcaagagaat15961agctggaacc caggaagcag aggttgcagt gagccacgat catcccactg cactccagcc16021tgggcaatgg agtgagactc catctcaaaa aaaaaaaaga tactcaggaa aaaaacgaag16081aaagaacaag ataacaaaat actgattgaa gctgagtgct ctataggtgc aggtgctcat16141tcactattct attttcttgt gtgtttgaac actttttttt ttttttgaaa tggagtttcc16201ttcttgttgc ccaggctgga gtgcaatggc atgatctcgg gtctcaactc actgtagcct16261ccgcctccca agttcaagtg attctcctgc ctcagcctcc caagtagctg ggattacagg16321cgcccgccat gacgcccagc taattttttt tgtattttta gtagagatgR ggtttcgcca16381cgttggctag gctggcctca aactcctggc ttcagctgat ccacccgcct cggcctccca16441aagtgctgag atcacaggcg tgagccaccg cacctggcct cccagagcgc tgggatcaca16501ggcgtgagcc accacgccca gcctgtttga acacttttgt gacgttttgt tgtttgtttg16561ttttgagatg gagtcttgct ctgttgccca ggctggagtg cagtggcaca atcttggctc16621attgtgactt ccgcctcccg ggttcaagca attcttctgc ctcagcctcc cgagtagctg16681Rgactacagg tgtgcgccac catgcctggg taatttttgt atttttagta gagacggggt16741ttcaccatgt tggtcaggct ggtctcaaac tcctgacttt gtgaaccacc cgcctcagcc16801tcccaaagtg ccagtgttac aggtgtgagc caccacgccc ggcctgtaac attttcatga16861aggtagagta tacttaattg caagaaaatt ttgaaaaaga agagaaatgg aacacttcKa16921ttggctatta aaaaatacca tgctgagtga aaaaagccaa tcccagaagg tcacatgctg16981tatgattcca tttctttttt tttttttttt aggctggagt gcaaatggca cgatcttggc17041tcactgcaac cttggtctcc cgggttcaag caattctacc tcggcctccg gagtagctgg17101gactacaggc atgcgccacc atgcccagct aatctttgta tttttagtag cgatggggtt17161tctccatgtt ggtcaggctg gtctcaaact cccgacctca ggtgatccgc ttgcctcaaa17221ctcccgatct caggtgatcc gcttgcctcg gcctcctgaa atgctaggat tacaggcgtg17281agccaccata cccggccgtg attccatttc tgtaacattc tctgaatgac aaaattatag17341acagggagga cagattgttg attgctaggg ccttggcgtt ggggtaggga gtgagggggt17401ctgtgaaggg cagggctttg gggttggggt agggagtggg tgtgtctgaa gggcagcagg17461aggaagatgg gattgttggt tttggtggtg attgattaaa tggcgtagct gtacatacac17521tcatagtacc ggagtccatt tcctcgtgtc gataatggta ctatagttat ataaaacgtt17581cgctttggga gccaagcaca gtggctcacg cctgtaattt cagaactttg ggaagcctag17641gcaggcagat cacttgagYc cagaagttca agaccagcct gggcaacatg gtgaaacctg17701atttttacaa aaaagacaag aattagctgg gtgtagtgga gtgtgcctgt agccccaact17761aatcaggggc ctgagatagg aggatccctt gaccccatga ggttggggct gcagtgagcc17821aagatcgtgc cactgccctc cagccccggc gacagagtaa gaccctattt atgcttccta17881gcaaggaaag acagtacaca tattgaagaa tttgactgcg ttaaaatgtt aaacctctcc17941ccaaactctt ataaacaatt gtagaacaaa tgataagctc aaaaaaatat ttacatctat18001gagaggccaa agaatatcag ttgactctgg atcaggggct cggacacgtt gctgtggagg18061cacacatctt cctggaaggc ggtttggtaa caaggtggca ggactcaaga cagcccacac18121gatccctgcc cctgggaaac agccatgtgt aaccctcccc tcgagtgtgg gYggaatgag18181gaagagggag gtgtgtctct cctgcgatta gctcactcat ccattgagtt tgacgtcatc18241aaaatgcgag atgatcctgg atgggcccga cctgatcagg gaagccagac agtaaaagtg18301tcagaggctc tttagggccc gacctgatca gggaagccag acagtaaaag tgtcagaggc18361tctttagggc ccgacctgat cagggaagcc agacagtaaa agtgtcagag gctctttagg18421gcccgacctg atcagggaag ccagacagta aaagtgtcag aggcttttta gggcagcgag18481aaaacgggaa cttcattcct tgaaccggga ataaattctg cctccaaccR gaatgggctt18541agaagagggt cctccagctc ctcatgagaa tgcagtgcag ccggcccctg gaattcagcc18601tgcgtcccag ccacgctgca cctgcacccc tgacgacgaa tgcagtaaga tgataagtgg18661atgtgctggt ttcagccctg aagttggtgg tgatttgttg tgcagcaata gaagtacaca18721caatatgtac caagaacctt gagaagagtc acatttattc tttctgtttt gtttctgaga18781tggtctcacc caggctggag tgcagtggca ccatcttggc tcattgcact cttgacgttc18841tgggctcaaa cgattctccc acctcagctc cctgagtagc tggggccaca gtgcacgcca18901ccacccccag ctattttttg tagtttttgt agagatgcgg ttttgtcatg ttgcccaggc18961tggtctcgaa ctccagaggc caagcaatct gcctgcttca gcctcccaag gtgctgggat19021tacaggcgtg agccaccgca cctggcccaa gattaatatt tctttttttt tttgagatgg19081agtcttgctc tgtcgccacc caggctggac tgcaatggtg catctctgct cactgcaagc19141tccgcctccc gggttcatgc cattctcctg tctcagcctc ccgcatagct gggactacag19201gtgcctgcca ccacgcctgg ctaatttttt gtattttttt tttttttaag tagagatggg19261gtttcactat gttaaccaga atggtcttaa tttcctgacc tcgtgatccg cccgcatcgg19321cctcccaagg tgctgggatt acaggcgtga gccactgcgt ctggcctaat atttctaaat19381gaagaaacta tactYtgagg aagatgaaat aagtcttgtt ttagactaga aaatttcaat19441agtgtaaata cgttaacttc tgtaaaatca gtcaacacca agtcaaaaca caaaagatct19501tttgggaata tgagcagatt attcaaaggt tcatctaaag aataaaaatg aggccaagtg19561tagtgattca cgcctgtaat ccYggcactt tggaggccga ggcaggtgga tcacttgagg19621tcaggagttc aagaccagcc tggccaacat gatgaaagcc catctctact aaaaatacaa19681aaattagcta ggcgtggtga tgggtgcctg tagtgccagc tacttgggga ggctggggca19741ggagaatcac ttgaacctgg gaggtagagg ttgtagtgag ccaaaattgt gccactgcac19801tccagcctgg gcgacaagag tgagaatcca tctcaaaata aataaatgaa taaaaatgaa19861agcagtagga aacatctgga aaaccagagg catgaggggg tcgggccctc tgatactcaa19921gtaggttaga gtaaaaaaaa gccagatatg gtgatgtggg aatatatggt cccataggtg19981ggatctgtgg ttccagctac tcagtgaggt ccgaggatca cttgggcctg ggaggttagg20041actgtagttt gctatgatca catccacgaa tagccactgc gctccttcct gggctacaga20101gtgagaccct tctctaaaaa tggaagaaaa tggccggcac agtggttcat ggctgctgta20161atcccagcgc tttgggaggg tgaagtgggagaataacttg agcctgagag ttcgagacca20221gcctgggcaa catggcaaaa ccgtgtctct acaaaaaatt tcaaaattag gtgggtgtgg20281tggtgtgtag ggtccagccc cacagggtcg gtgggttttc tccccatgtg cggagatgag20341agagcgtaga aataaagaca caaggcaaag agataaaaga cagctgggcc cgggggacca20401ctaccaccaa gccgtggaga ccggtagtgg ccccaaatgc caggctgcac tgatatttat20461tggatacaag acaaaggggc agggttagga gtgtgagcca tctccaatga taggtgaggt20521catgtgggtc atgtgtccac tggacagggg gcccttccct gcctggcagc caaggcagag20581agagaggaga cagagagaca gcttacgccg ttatttcagc atatcagaga cttttagtac20641tttcactaat ttgctactgc tatctaaaag gcagagccag gtgtacagga tggaacatga20701aggtggacta ggagcgtgac cactgaagca cagcatcaca gggagacgat taggcctctg20761gataactgcg ggcaagcctg actgatgtca ggccctccac aagaggtgga ggagcagagt20821tttctctaaa ctcccctggg gaaagggaga ctccctttcc tggtctgcta agtagtgggt20881gtttttcctt gacactgacg ctactgctag accatggtct gcttggcaac gggcgtccta20941ccagacgctg gcgttaccRc tagaccaagg agccctctgg tggccctgtc tgggcataac21001aaaaggctca cactcttgtc ttctggtcac ttctcaccat gtccccccag ctcctgtctc21061tgtatgacct ggtttttcct aggttatgat tgtagagcga ggattattat aatattggaa21121taaagagtaa ttgctacaaa ttaattaatg atattcatat ataatcatgt ctgtgatcta21181gatctagtat aactattctt attttatata ttttattaca ctggaacagc tcgtgccctc21241ggtctcttgc ctcggcacct gggtgacttg Ycgcccacag tggtgcgtgc tggtggtccc21301agctacttgg aaggctgaag tgggaggatc atttgagccc aggaagttga ggcttcagtg21361agccatgatc atgccacagc atttcagcct gggccacaga ccttcaagat cctgtctcaa21421aaagaaaaag ggRtatcKct cagtgcttag tgcagctgtg ggtcattcgg cagctgacac21481agatgaggct atgtaacgca gcactgcagg cagaggggga ggccaaggcc tgaatacccc21541gtctgactag ccttaggcag gcgacttggc ctctctgcct cccccatctg gttagccttg21601ggcagtcgag ttggcctctg tgtctcccct gtttgactag ccttgggcag atggcttggc21661ttgtctgtac ctcagttttg tctgaaatat gggYtgtcag agacetatag cagaagctgc21721tgtgtgtcat tattcactgt tctagggctc agaacaggca cgcgataaaa gccagcaccg21781ttggccgggc gcattggctc acgcctgtaa tcccagcact ttgggaggcc aaggcaggta21841gatcacctga ggtcgggagt tcaagaccag cctggccaac atggagaaac cctgtctcta21901ctaataatac aaaaattccc caggtgtggt ggcaggagcc tgtagtccca gttgctcagg21961aggctgaggc aggagaatca cttgaactca ggaggtggag gttgcagtga gccgagatca22021caccactgca ctccagcctg ggtgacagag cgagactccg tctcaaaaac taagaagaag22081attaataata aaccagcact gccattgcca gcagtccatc ctacctcgca tccaggttcg22141tgagctacca agtgtcccca caaagctgta aaacaRcatg accactgtag ttacgtttct22201gtaaaaaggt gactgagtct tctaaccctg accacacata tgtgtgcaga gagagccagc22261tccatcctgc tctcccagaa gcctgaacag ctgtgcccta gctcgtagca gtggcactcc22321aggaaggggg cgggcggctg tgcccgagct cgtagcagtg gcactccagg aagggggcgg22381gcggctgtgt ctgagctcgt agcagtggca ctccacgaag ggggtgggcg gctgtgcccg22441agcttgtagc agtggcactc caggaagggg cggccggcca gccaggagag cagccggtga22501aggagcactt ttgctttgta cgtggcaacg ctcggcaggc tcagcagttt cacaggtctc22561aggcgttcct tgtaacctta acaggtttaa gaaaatgtga gcaacatcag acatgtccat22621ttctagaaca ttctaatgga tgaagttctg tacagagggc tttagcagaa ttagagacct22681ttggagtgga aatgatgtct ctgaaccttg agctctgagg cggttcctgc ctggacgtgt22741gccctccctc cttcctttgc tggtcgccct gacaccatcc tgccacggat agtggaagtg22801tttctcctgt tttcttccct gaaaaagggt gtccagcatc agcaaccttc atggctgagt22861tgcacatccc ttaagccctg taatgttcat ggacagtttt gatcactttt ggactcaccc22921attttttctg ctaaccatac atcttaattc ctttactctg accaatggca ggaggtgtgt22981gcatcgtgga aggagctttc cacacatcta ctaaatgcct ggatgttttt tcctctagag23041aaacctgatg taaagcaaaa gtccagcagg aagaaagtgg tcgttccaca gatcatcatc23101acgcgagcgt cgaatgagac gctagtcagt tgcagttcca gcgggagtga ccagcagaga23161accattcggg agccggagga ctggggcccc taccggcggc acaggaaccc cagtacagca23221gacgcctata attcacatct caaagaataa acaagcacct ttgcatggca ctcgctactc23281cctgcgatag gcgtctcggg aacagccgcc tcaccctgtg agaagccgag gccccttctc23341cagtgctctc ggggaggttg caccaggccc accccaccct gtgagaaact gcagccccct23401tctccagtgc tttcggggag ggtgcaccag gcccgcccca ccttgtgaga agccgtggcc23461cccttctcca gtgctctcag ggagggtgca ccaggcctgc ccccgccgtg agaaactgca23521gYccccttct ccagtgctct cggggagggt gcaccaggcc caccccaccc tgtgagaacc23581tgcagccccc ttctccaatg ctctcgggga gggtacacca gggttgcccc accctgtgag23641aagccaccgc ccccttctcc agtgctctcg gggagggtgc accagggctg ccccacgctg23701taagaagccg ccgccccctt ctccagtgct ctccagcccg ccccaccctg tgagaagcca23761tggccccttc tccagtgctt ttggggaggg tgcaccaggg ccaccccacc ctgtgagaag23821ccgtggcccc cttctccagt gctttcgggg agggtacacc aggcccgccc caccctgtga23881gaagccgtgg cccccttctc ccgtgctctc agggagggtg caccaggcct gcccccgcct23941tgagaaactg cagccccctt ctccagtgct ctgggggagg gtgcaccagg cctgcccacc24001ctgtgagaaa ctgcaccccc ttctccagtg ctctcgggga gggtgcacca ggcccgcccc24061accctgtgag aagccgcagc ccgttctcca ttgccatttg gtcctgcctt ccccagagct24121gctagcagct cggtcgaggc aagagtgact ggctcagaga gcgagctttg tggtattgat24181aaatgaataa attagtatgt tcatgttgga agtggctggc tctgcagagt gatggacagc24241aggtggcatc ttacattgtg atttgtggtg cacagttaaa acacttccct tcgtttcaga24301aggagacaat gtgaatactt ggaaattata gtgaataaga tgcctcccaa acttgaggaa24361aaggtcagat tctaagagct aaacgttaag atgtaatgtt ccagaacaaa gaggcccaga24421gtcactgaac ggggccccag ggctcctagc ttgagtgtaa tctgagtcag cttatgaaaa24481ggccccaaag aagaaagtgg aatgtgtgct gggcacatgg gacagccatg gcactgagac24541tttggaagtg gccttgccag cgctgtcctg tctttctggc ccacagatga gggctcctga24601ttgtggaagg ccacggaagg gacctgaaca cgatggcacg agacaagtta tctcctgtga24661ctgctcctct ccagcaaccc tcagctcagg ctagactttc cccctgtgtt gagaatgccc24721tgccctgtgt ggaagcctga tgcatttagt gacagtgtgt tagctttgtt agtttctagt24781ctggccagca ggctttccat gtgatctgcc attggcttgc ttcagtggcc agtggagcca24841ggggtgtcca ccagagtctt ggacacagac ttctgaggtg gcagcgggag ggagcctgct24901ccctgccaca cacgtacaat ttctaatcca tacaggccta tgtagtatat acacacatac24961gcagagcctt aggaattgtg aaaggccaat caaaacttgg ggccctaatt tcactctgcc25021aaaagaaaaa aaaaaaaagc tgaaagatga gtcacccaag aaactgcttc ctttttgttc25081ctaagcagag agctaccaat aaaaagttaa agtttctcca taggtagcta ctctgttcac25141cttctgtgaa gtgccaaatt ttttttttat tattatactt taagttttag ggtacatgtg25201cacaacgtgc aggtttgtta catatgcata catacgccat gttggtgtgc tgcacccatt25261aactcgtcat ttacattagg tatttctcct aatgctttcc ctcccccctc ccccctaccc25321cacgacaggc cccagtgtgt gattttcccc ttcctgtgtc caagtgttct cattgttcag25381ttcccaccta tgagtgagaa catgcggtgt ttggtttatt ttacttgcga tagtttgctg25441agaatgatgg tttccagctt cacccatgtc cctgcaaagg acatgaactc atcctttttt25501atggctgcat agtattccat ggtgtatatg tgccacattt gcttaatcca gtctatcact25561gatgggcatt tgggttggtt ctaagtcttt gctattgtga atagtgccgc attaaacata25621ggtgtgcatg tgtctttata gcagcatgat ttataatcct ttgggtatat acccagtaat25681gggatggctg agtcaaatgg tatttctagt tctagatccc tgaggaatcg ccacactgtc25741ttccacagtg gttgaactag tttacagtcc caccaacagt gtaaaagtgt tcctatttct25801ccccatcctc tccagcacct gtttcctgac tttttaatga tcgccattct aactggtgtg25861agatggtatc tcattgtggt tttgatttgc atttctctga tggccagtga tgatgagcat25921tttttcatgt gtctgtaggc tgcataaatg tcttcctttg agaagtgtct gttcatatcc25981ttcacccact ccatcaaaaa gtgggtgaag tgccaattta ctgagcacga gacaaataca26041tcattgactt cttttgtctt gtggattcaa ggatgtgtcc acgcccgccc tccttcccct26101ccagctcctc cccttgaaat actgaagccc tcaacatcat ctttggagga aggtacagag26161ttggctctca cgcgcacatc cttaaccttg gcctaataaa cctctaaggt ggctgagacc26221tgtctcagat acttgttggc ttacaaattg accaccatgc agggactcag agtggaggtg26281gccctgccct ttggcaggtt tcctcctggt gcttaggacc agtttgggcc ttctttattg26341ctcaggacag taggacaatt tgctgagatc cgggagccct accaccctcc agagaatctc26401tgatctcccg aaacttggtt gaggtcgaaa gtttattttg cgcacaatgc cttttccttt26461tccggtgttt tacttgcttc caacaaagaa ggcgagtttt cctgcttccg tgatgatggg26521gagcaggaga cttccctgga gtttcagctc ggaaacaact cgagtttttc ctgattccac26581gatgggggag agcagtctaa agcctgggcc ccattcctag gtaagtggct gaattggggt26641tctgtcttgg aagttctcct taatgactaa agctaacatt aacgactggc tggtcttaac26701ttcttactat tagcatgctc agtaagtgta taaattgtgg gattgtttgt tttgcttaac26761tgctttttgt ttgtttctgt gtttgctatt tttgctgttt cagtcttttt cccgttggat26821ttgacgaatt ctgttagact tggtcaaatc tgaagcaaac ttccaagttg tgggaaacaa26881gcctctgaat tagctgaatt cccacagctg ggaaaaagga aaaaaagcca gaaaaaggga26941aaaaagattt tgaccacgtg atgggcttta tttgcataac gaggccacgt ttttccagcc27001aagccacact cgaagagtaa gtgccgtcac ctcacgctgc agttcggtag gtagagtccg27061ccatttttcc accaggacag cccgggtttg gttcctacat cctttctggt ttgaaatttg27121tgttactttt gaaatatcag cagtttgtcc cggctgaaat aaggtagtga gatttaaaga27181tttaaaagga ttttcttttt ttttgagacg gagtctcgct ctgtcgccca ggctggagtg27241cagtggcacg atcccagctc actgcaacct ctgccgccca ggttcagcca ttctcctgcc27301tcagcctccc aagtagctgg gattacaggc acttgccacc gcacctggct aatttttgca27361tttttagtag agacggggtt tcaccacctt gaccaggctg gtcatgaact cctgatcttg27421tgatccaccc tccttggcct cccaaactgc tgggattaca ggcgtgagcc actacgccca27481gccaagagga tttttaaaag ggcttaatgg tcaaaagtaa acttaactaa aagctaatat27541cggatatatt ccaacatagt agcttaatta agtccgtatg ccacatacac atatacagac27601ccctgtgtag aaaagaaggc accagactcS tttttgggga gaaacttctg tttttcttat27661ggaactccaa gagtggaaac agacacgttc ctctcagatc ttaaattgct tgctattgta27721ttgtgttacc tgattttttt ttactaaaat agttgacaca ggactttttg gggtgctgct27781tctctagctg gaggcctctg tggccggtgg tgcccctacc cgggtctcct gggctccagg27841gttggccctg gagtaccccg cccacctagc ccggccgtgg gatccgccct cagcccatgg27901ctggactgcg catgcgcacg ccgcaacccg cttcagcctt gagcactgga gtcYggacaa27961gaggagtgtg gtggcgcaca aaacgcccaa agatgccagg aacggcagag ccccaagggt28021gtcacagctc gattgttccc cccgcctgcg gtctggtgag tgggggtgtg ttacagtgtc28081accccccctt tttttttttt tgagacagag tctcactctg tcactaggcc agagtgcagt28141ggcacagtct cggctcactg caacctccgc ctcctgggtt caagcgattc tcctacctca28201gcctcccaag tagctgggag tacaggtgcc tgccaccaca cctggctaat tttttttttt28261tttttttttt ttttttttga gacagagttt ctctcttgtt gcccaggctg gagtacaatg28321gcacgatcgc agctcactgc aacctccacc tcccaggttc aagcgattct cctgccttag28381cctcctgagt agctggaatt acaggcatgc accaccatgc ccggctaatt ttttgtattt28441ttagtagaga cagggtttct ccatgttggt caggctaatc ttgaactccc gaccttaggt28501gatccacccg cctcagcctc ccaaagtgct aggattacag gcatgagcca ccacacccag28561ccagtttttg tgttttcagt agagatgggg ttttaccatg ttggccagga tggtcttgat28621ctcttgacct tgtgatccgc tcacgtcagc ctcccaaagt gctgggatta caggtgtgag28681ccaccaggcc tggccgtcac cccctttttg aatctgtgat tcagcaagtc ccaaatgctt28741ctcccatgtc caagaaaaat gaaggtatgc agacaaccgg agggtgagca ggacaacgtt28801ttgttgagca acagaacagt tctcagtgaa gaggggagcc aaagtgggca gcccctactc28861aaagttgggt acgcccccac tgaagtcccc caccatgcac agcctataat tttatctttg28921acttggcatt cttttttttt tttttgagac agagtcttgc tctgttgccc aggccggagt28981gcagtggtgc gatctcggct cgctgcaacc tccgcctcca gggatcaagt gttctcctgc29041ctcagcctcc tgagtagctg ggattacagg tgcccgccac tacacctggc taattttttg29101tttgtttgtt tgtttgtttt ttgagacaga gtcttgctct gttgcccagg ctggagtgca29161gcagcgcaat ctctgctcac tgcaagctcc acctcccggg ttcacgccat tctccggcct29221cagcctcccc agtagatggg actacaggcg cccaccacca cgcccagcta attttttgta29281tttttattag agatggggtt tcaccgtgtt agccaggatg gtctcaatct cctgaccaca29341tgatccgccc acctcggcct cccaaagtgc tgggattaca ggcgtgagcc actgcgccca29401gcctaatttt tgtattttta atagagacag ggtttcacta tgttggtcag gctggtcttg29461aactcctgac ctcatgatct gcctgcctcg gcctcccaaa gttctgggat tacaggtaac29521attcattttt aatgtccctc taacacaccc agactctccc tctgtatttt gagatgtaaa29581ttttgctatc tgatttttca cctaagagtt cttcctttgg tatgtcaatt tagggctatc29641aagctgaaaa ttgcctaaga caatgaaaca ggttatcaag aaattggaag tttaggccgg29701gcgcaatggt tcacgcctgt aatcccaaca aattgggagg ccaaggcggg cagatcactt29761gaggtcagga gttcgagacc agcctggcca agatggtgaa accccatctt tactaaaaat29821acaaaaatta gctgggtgtg gtggcacgta cctgtaatcc cagctacttg ggaggctgag29881gcaggagaat cgcttgaacc tgggaggcgg aggttgcagt gagctgagat tgcgccattg29941tactccagcc tgggcaacaa gagcaaaact ccatcttaaa aaaaaaaaaa tacaaaaatt30001agccgggcgt ggtggcaggt gcctgtaatc ccagctaccc aggaggctga ggcaggagaa30061tctcttgaac ccaagaggtg gaggttgcag tgagttgaga ctgtgccact gtactccagc30121ctgggtgaca aagtgagact ccaactcaaa aaaaaaaaga aaaagatcga aagaaagaaa30181ttgtaagttt aaaatagtag aaaaaaaaga aggggtctta tgcatcttct gtgtgtcgaa30241tgcatctatg tatctatgag tcatgtatac aatgtttcac taccaaaaat atataaaaga30301gttttaatta attgggttaa aggaaaaaaa agcacttaaa tactgtatca gaaaaatata30361aacttttttt tttttttttt tttttggaga cagaatgttg ctctatcacc caggctggag30421tgcaggggca cgatctcggc tcactgcaag ctccgcctcc agtgttctca cgccattctc30481ctgcctcagc ctcccgagta gctgggacta caggagcccg ccaccacgcc cggctaattt30541tttgtatttt tagtagagac ggcgtttcac cgtgttagcc aggatggtct cgatctcctg30601accttgtgat ccccccgcct cagcctccca aagtgctggg attacaggtg tgagccacca30661tgcctggcca gaaaaataga aactttaagc ccaaatgctt tttcaggttc atgtgactta30721agtaaatcct taaaaaataa gctagtttta aaattattgg taaaataaaa tagaaatgcc30781ttcagaattg tcaacataca ttattgttta gatttattgg taaaggagtt caatatttat30841cattacgaga tactgtaagt tgtcaaaatt tggcatgagg gttataaggc ttgcaacaca30901aaagggaatt atgtttgact aatttcttga taaataaggc atttaatatt gtttaatgga30961aagagataaa tcctgagttg ctggcaaaaa aaaaaagagg aacatttatt taaccataag31021gttcttactt aggtaaacac ctaaaattca caggctataa aaatggttag gccaggtatg31081gtggctcacg tctgtaatcc cagcaatttg ggagtctgag gtgggcagat cacctcagat31141caggaattca agaccagcct ggccaacatg gtaaaacccc gtctctacta aaaatacaaa31201aattagctgg gcatggtggc aggtgcctgt aatcccagct acttgggagg ctgaggcagc31261agaatcactt gaacccagga ggcggaggtt gcagtgagcc aagatcacac cattgtactc31321caacctgggc aaaaagatca aaagaaaact ccatctcaga aaaaaaaaaa aaaagaaaat31381ggttaacaga gaaataactt taaatgaaga ctatcacagt tttctttcct ttttcttttt31441tttttttgag aagagttcac tctgtcaccc aagctcgaac gcaatggcgt gatcttggct31501caccgcagtc tccgcctccc aggttcaagc tattctccca cctcagcctc ccaagtagct31561gggattatag gtgcccacaa ccacgcctgg ctaatttttt tttttttttt ttttgagaca31621gagtctcgct ctgtctccca ggctggagtg cagtggtgct atctcggctc actgcaagct31681ccgcctcccg ggttcatgcc attctcctgc ctcagcctcc tgagtagctg ggactacagg31741cacctgccac cacgcctggc taattttttg tatttttaat agagatgggg tttcaccgtg31801ttagccagga tggtcttgat ctcctgacct tgtgatccgc ccagtctggc taatttaaaa31861aaaatatttt tatagtaggc tgaggcgtga gaatggcatg aatctgggag gcggagcttg31921cagtgagccg agactatgcc actgcactcc agcctgggca acagagtgag acaccatctc31981aaaaaaaaat ttattttaaa taaacaaata aataattaca gatgtaaaga tagttataag32041gagatatttt tagtatgaaa agttaaaagg aaaataattt catatgagaa agaatcttgt32101gtggtaaatt tttgttctaa aatggctagt tattttggaa agagggatgt tcagaataaa32161acaggaagtc caagtatatc ataaatggtt tgtgtaagtt ttgataaagt ttgtaaaaag32221cttttttttt ttttttttaa gatggagtct cgctttgttg tctgggctgg agtacagtgg32281tgcaattttg gcctatgtca gattaacaag gctttcttgg agcattaacc cactctttaa32341ttaaaaaaaa aaagttataa aaatgtttat ggaaattatc ttttttgttt ttgaggcaga32401gtttcgctct tgttgcctaa gttggagtac agtggtgtga tctcagctca ctgcaacctc32461tgcctcctag gttcaagcaa ttcttctggc tcaggctccc aagtagctga gattacaggc32521gcccacaacc atgcctggct aatttttgta tttttggtag atacgtggtt tctccatgtt32581ggccaggctg gtctcgaact cctgacctca ggtgatcgac ctgcctcggc ctcccaaagt32641gctgagatta caggtgtgag ccaccgcacc cagccttatg gccttatttt ttttgttttt32701ttttagatgg agtcttgttc tgtcacccag gctgaagtgc agtggtgtga tctcggctca32761ctgcaacctc cacctcccag gttcaagcag ttctcctggc tcagcctccc aagtagctgg32821gatcacagac gcacactacc atgcctggct actttttgta tttttaggaa agatgaggtt32881tgccatgttg gtcaggctgg tcttgaactc ctgacctcaa gtgatccgcc tgccttggcc32941tcccaaagta etaggattac aggcgtgagc caccacaccc agcctgaaaa ttatatcttg33001tgctcaagat gattacaatt taatagattt gtttataaaa ttttagagag atttaattgg33061ccttatgctg tctttatctt ttgagaagaa attccctcaa agaataaagg tttctacatt33121tttttttgaa atctttgagt tatctcagca ccataggaaa ggtaaaaaca gaaaaaaaaa33181aagaaaataa aaaaagaaat ctgtgagtta tcactgtggc taagtgaatg acttatttta33241caatgacctg tgatcctatt tagtgatagc aactgtttta aaactttttc taggccaggc33301acggtggctc acacctgtaa tcccagcact ttgggaggct gagatgggca aatcaagagg33361tcaggaaatc aagaccatcc taactaacat ggtgaaaccc tgtctctact gaaaatacaa33421aaaattagct gggcgtggtt gcaggcgcct gtagtctcag ctgcttggga gactgaggca33481ggagaatggc gtgaacccgg gaggcagagc ttgcagtgag ccaagattgc agcactgcac33541tccagcctgg gtaacagagc gagactctgt ctcaaataaa aacaaaaaca aaacaaaaca33601ctttttatat ttgacaaact tKccaaaatc aaattctttc ttcagtcctc attaattttt33661gtgtgtgtgt gtctgatgga gtctcactct gttgccccag ctggagtgca gtggcacaat33721ctcggctcac ttcaacctcc acctcccagg ttcaagcgat tctcctgctt cagcctccca33781agtagctgga actacaggtg cccgccacca cacctggcta atttttatat atttttttct33841ttttgagata gagtcttgct ctgtcaccaa ggctggagtg cagtggcacg atctcagctc33901actgcaagct ccgcctcccg ggttcatacc attctcctgt ctcagtctct tgagtagctg33961ggactacagg cgcctgccac cacgcccagc taattttttt tttttgtatt ttttagttga34021gacagggttt cactgtgtta gccaggatag tctccatctc ctgacctcat gatccacctg34081cctcagcttc ccaaagtgct gggattacag gcgtgagtca cctgcctcag cctcccacag34141tgctgggatt acaggtgtga gccactgcgc caggcctccc aaggtgttgg gattacaggc34201gtgagccacc gctccgggcc tcccacagtg ctaggattac aggtgtSagc tgctgcacct34261ggcaattttt tgatattagg tcccctgaag tccaaaaaga gatatatggc ttatttggta34321taatgaaatc atacaggaag cattgttaaa ggtgaaatgg tgtttttctt gggattatat34381ttatataaat gtgttgttag tgtgtgttcc aaaattgtat gagatgcctg tgattctgat34441acatcttagt atatggtatt agtagtaatt atgattatta tgtaaaattg ttgtatgcca34501cataagtaac caaatttgct tgtcaattat gtcttttttt tttctttttt ctgagacaga34561gtcttgctct gtggcccagg ctggagtgca gtggtacagt cttggctcac tgcaacctct34621gcctcccagg ttcaagcgat tctcatacct cagcctccag agtagctgga ctgcaggcgc34681atgctaccac ggccagataa tttttgtatt tttaatagag acggggtttc tttttttttt34741ttttccctga cagagttttg cttttgttgc ccaggcttgg agtgcaatgg cgcgatcttg34801gctcactgca acctccactt cctgggttca agcaattctt ctgcctcagc ctcccaagta34861gctgggatta caggtgtaca ccaccatgcc tggctaattt tgcatttttt tagtagagac34921atggtttcac catgttgatc agggtcgtct caaactcctg acctcaagtg atccacccgc34981ctcagtctcc caaagttttg ggattataaa tgtgcgccac tgcacccggt caagacaggg35041ttttgccatg ttagccaggg aggtctcaaa ctcctgacct caagttatcc gccctgcttg35101gtctcccaaa gtgctgggat tgcacacatg agccactgtg cctggcctca attatgtctt35161taactgtggc tgttctaaga cttttgtcat ccacaattgt tgttttactt ttatcctttc35221acagagcagt ttataatcag ctatagaact ctgaggtact cttcaatcta agtttctgat35281cacttttgag attgtgccat tggaatagag caaaaaactt ccaagactct cattgagagc35341tgctatattc atgaggattg ttcatccaat attgggcaaa atgagagtta attttatgga35401ctaaactaat ataagactga aataatcttt ttatgttttt gcttaaaaca ttactgattc35461tttctgttct ttcagaacca agaaaacttt tttttttttt tttttgagac agaatctcac35521tctgtcgccc aggctggagt gcagtggcgc catctcggct cactgcaagc tccgcctccc35581aggttcacac cattctcctg cttcagcctc ccaagtagct gggacztacag gcgcccacca35641ccgcgcccgg ctaatttttt gtatttttag tagagacagg gtttcaccgt gttagccagg35701atggtctcaa tctcctgacc ttgtgatccg cccacctcgg cctcccagag tgctgggatt35761acaggcgtga gccacggtgc ccggctggta acttttttaa ctttgagcta tttgcagctt35821ttagcaattg agtaaagtac actcctgtga ggaagatttg gaacatattt ctgttttctt35881ttttgttttt tatattttgc ttgggtagca tatttctttc tgatttctcc aaaaattggg35941aactatctgt gagcactatt taaaaaaaaa aaaaaaactt ttccctctcc cttcccctat36001ttgtattcta atgtcaatat aatgatgtgt gcacgtgcag taacaatctg ttttctcttg36061taataggaca tagagacact gcatatttta ccaaggcttt gactggaatg gcatgctttc36121aaatataaac agacggcttt aaggaatcag agttgaccta tagtgcccat caaagccctt36181gggaaaactg gcttcatact gtatccaccc agtggattct tcttgcccag atggagccca36241tgtatcaagg tagggaaact gcaatagaga aagggtttaa ttcacacaga gacaagtgaa36301tgggagacca gaatttgatg attactgaaa tcagtctccc caagaatttg gggactaggg36361tgttccaaag gtagtttggg ggaaggggtg gggtagcttg gcagtgggtg cttgctgctg36421atgggttgtg ggggcggggg gccatcgtag ggtgtgagac aggatcctcc cgtgtgctga36481accgcttctg ggtggggctg caggagtagt tggcaggtcc aggtggagcc atcatcagtc36541agtgggtcct gttggagccg taggtatcag acatgcaaaa aacccttgSg aggtactgca36601aaaggccaat cttaggttac aatagcattg ttatctgcag gacctctgga ataatggctg36661ataattgttt atgtctacat cttagccaaa ttcaggctcc tctcctctcc ctagcctggt36721ggcccctcat tagttttaca aatacagttg agttttgggg aggggctatt atcatgtaaa36781ctataaacat ctcccaaagc tagcccaaga ataactgaag gcagcttgaa ggctaaaggt36841caaagggggt ttggctacat cagatcttcc ccaattggca taattttctc aatgatacaa36901tttttgtggc cgggctcggt ggctcacgcc tgtaatccca gcactttggg aggccaaggc36961gggcagatca cgaagtcagg agattgcaac catccttgtc aacacagtga aaccccgtct37021ctactaaaaa tacacaaaaa attagccggg cgtggtggcg ggcgtctgta gtcccagcta37081cttgggaggt taaggcagga gaatggcgtg aacctgggag gcggagcttg cagtgagccg37141aggtcgcacc actgcactcc agcctgggcg acagagcaag actccgtctc aaaaaaaaaa37201aaaaaaaaaa aagcctacat ggtaaatatt tattcttgct acagtttata caaataacca37261ggccaagtac aataagacta aaacttattt tgaaaacaaa tcagtactac ctgatttcac37321tttggtaaaa ataggagact agagagagaa aaattatgtt tcaaaataaa ctatagtata37381cctgttatta gattctagcc ttgcctaagg tttttcaact tttattattt tctctttttt37441tgtttgttta tttgtttttt cagacagggt ctcactctgt cacccgggca gtggtgtggt37501atcagttcac tgcaacctct gcctcctggg ttcaagtgat tctcgtgcct cagcctcccg37561agtagctggg attattggtg tgcaccacta tggccagctc atttttttgt atttttagta37621gagatgcgtt tcaccatgtt ggccaggcta gtctcgaact cctgacctca tgatccaccg37681gcctcggcct cccaaagtgc tgggattata ggcatgagcc actgcgccgg gccaaagccg37741tgcttttctt aaagctctgc gaacgaaagc tagacaactt aaacctcaga agagaataat37801ggtacctatt tatatacata aaccactttc acgcctgcct gctgtgtatg gacttcagag37861gaacacagcc tacgtcagtt ttctaggatt gttctctcca cgtgctcttt gttttctttc37921tttcttcttc cctctatttt cctccctccc tcccttcctt ccttcctttc acaaagtctc37981actctgttac ccaggctgga gtgcagtggc acgattttgg ctcactgcaa cctccgcctt38041ccgggttcaa gcgattctcc tgcctcagcc tcccgagtac ctgggactac aggcaaccac38101caccacaccc agctaatttt gttagtttta gtagagacgg ggtttcacca tgttggccag38161gctggtctcg aactcctgac ctcaagtgat ctgcctacct cggcctccca aagtgttggg38221attataggca tgagccactg tgcctggtca caacacctgc attttctttt ttttttgttt38281ttttttttga gacagagtct cactctgtca cccaggctgg agtgtagtgg tgtgatcttg38341gctcactgca agctccgccc cgtgggttca caccattctc ctgcctcagc ctcccgagta38401gctgcaacta caggcacccg ccaccatgcc tgcctaattt tttgtatttt ttagtggaga38461tggggtttca ctgtgttagc caggatggtc tctatctcct gacctcgtga tcgacccgcc38521tcagactccc aaagtgctgg gattacaggc atgagccacc acgcctggcc aacaccccca38581ttttgtatac cacactcaac tcccccattt tctacatgct gatcaaaacc aatctgaatg38641gagtctaggt ctggctgtgg gctgttggga aSaggccccc aaatctggcc atcaactggc38701cgcaaaactg gccataaaca aaatctctgc tgcactgtga catgctcatg atggccatga38761ctcccacact ggaatgaggg caaggaacac ctggcccacc cagggcggaa aaccgcttaa38821aggagttcct gaaccacaaa caatagcatg agcgatctgt gccttaagga cgtgctcctg38881ctgcagataa ctagccagac ccatcccttt atttcccgta agaagtgctt ttaatctata38941atctatagaa acaatgctta tcactggctt gctgtcaata aatatgtggg taaatctctg39001ttcgaggctt tcagcttcga aggctgtgag acccctgatt tcccactcca caccctatat39061ttctgtgtgt gtttctttaa ttcctctagc gccgctgggt tagggtctcc acgaccgagc39121tggtcccgca gtgggcagtg gtttgggccc gaccgagctg gtcccgcagt gggcggtggt39181ttgggcccga ccgagctggt cccgcagtgg gcggtggttt gggcccgacc gagctggtct39241cgcagtgggc ggtggtttgg gcccgaccga gctggtctcg cagtggtttg ggccctgctt39301gctcgactgg atttcaacag tgaagccaga ttagaggaca tgctggctgc cccgatcctg39361gacctggacc aatggtgctt cctaaacatt ccatgtcaac ttttcttttt ccaccttcca39421gggctgggct gtgaaagaga ttggagaggc aggtgcctgt gagagaagct tctaccagcc39481tgggaagtca ggtcttccat gagtgagggg aggtatctac tttgccaaac cttgttagRg39541aatgctgaca tagctgcctc ttcctgccca gaagctaagc ttgaatgcca cccctcacag39601agcttctctc ctgtctttca ctcactcctg cttatttctg tgacagaact tgacagcatg39661gacattgcca cttgcttcct tcagtctcct ggagtcccca tgtcctgaac acagaggagg39721cacccaaatg cttgctgaag ccacatccct ggaccaagtg acctgaccct cacctggagg39781accttcacct tcacctctca cctggaggac cctctcacct ggaagactct ctcacctgga39841ggaccRcccc acctgaagga ctcccaactg gaggaccctc ttacctgggg gaccccctcc39901gctggaggac cctctcacct ggaggaccct cttacctgga gggcccttca cctgggggac39961tcccccacct agaggatcct cttacctgga ggtcccctca cctggggaac ccccccccca40021cctagaggat cctcttacct ggagggccct tcacctgggg aacccccccc ccccacctag40081aggatcctct tacctggagg accccccacc gagaggaccc tcccccccca cctggaggac40141ctccccacat gaaggactcc cacctggagg accttcttac ctgggggaac ccgtctcacc40201tggaagacct tcacctgggt aatcgccgtg gcctcccact acggcgcagc cgggtcggct40261gcccgggctt caccctaaaa taaggcccga actgcagcag tcggagccga tccctggagc40321ccagcgccta gggcggaccc gcgggagcgt ctcttggtaa ccgttgctaa ggagaSgaag40381cccgggaagg agctttcgcg cctgcgccgc ggggccgtcc gcgtctgcgc ctgcgcgcaa40441gagaggcggg gccagcgctc ggcatggcgg agccagatct ggagtgcgag cagatccgtc40501tgaagtgtat tcgtaaggag ggcttcttca cggtgcctcc ggaacacagg gtgcgcgggg40561tgccacccgg gcagctctgc ccgcctcgct agcggcactg cccggctggg tctggggagc40621ctcgtgtcgc gctgccgcgc cgaggcttcc ggcacgggcg ggaacgacag tcccagagtt40681ccccgcggcg ggggcggaag ccggggcggg gcgggctcag gacccccgac agccggtccc40741tggagatctg aggggccggg gcgggctcag ggatgcctcg Mgcccgcgga gagacgggcc40801cgggacttgg gaagagcaag ctccgggatc cagctcgggc gctgctgggt tcagcgcccg40861agctgggctt tgcaggctga gccgcgagcc acttgtttgt ggggagaatt tacaYccgcg40921acgagttcga gcttgaaggc tcccgctggg cttgggctgc atggagcggg gtccagcgct40981gcctggcggc ttcacgcgaa ccctgggcgc gtcccttgcg gaggtgccgg tgcctccctc41041tgcagagacc aggagagagc ctctgggcgt tgggcggtgt tgcccaggtg cacggctggt41101gggggtggag agccctgaac tttggccgct ggtttgtgtt tttaactgct ggctggtgct41161ctgagaggcc aagccacgtg ttcagtaaga atcattaaca gatcgtggct ggatgacaac41221aggagecaca ctttactcat ccagacgtgc agcaggcgct caccgcgttg agagccgtgt41281gctggactct ggggtgaagt agtgagttgg ccttttcttt tttgttgttt ttttgagaca41341gggtctagcg ctctcgccca ggctggaatg cagtggcgtg atcacggctc agtgcatcgt41401cagcctcctg ggctcaagtg atcttcctgc ctcggcctcc caagtgctgg aattacaggc41461gtgagccacc gcccccagcc tggcctggca tttctttgag ttcaggaagt gtgacaagga41521tttggacacc cagaaataag cgtgtcgaga agagcacaag cagaggatgt gagaagggga41581attaaggcgg gaaagggagK ttgagactag gccatcagtg agtgacaggc cagatgcttc41641tgactcggtc cccaggtgtc aggaaggaat cagactgagg acaaatgcgg ccagtggctc41701acacctgtaa tctcagcaat ctgggaggct gaggtgggag gattgcttga gcccaggttt41761gatactgagg gttaagagaa gggaagccgg gaaatggggg ctgtgaggtt tcaaaccttg41821ctacgtgtgc tttttttttt tttttttttt ttgagacgga gtctcattct gtcacccagg41881ctggagtgca gtggcgcagt ctcggctcac tgcaagctcc acctcctggg ttcacgccat41941cctcctgcct cagcctccca agcagctggg actacaggcg cccgccacca cgcccagcta42001attttttcat atttttagta gagacggagt ttcaccgtSt tagtcagggt gctctagatc42061tcctgacttc gtgatccacc cacctcagcc tcctaaagtg ctgggattac aggcgtgagc42121cactgcgccc ggcttacatg tgcttcttag atccatggag aaggatacag actctactag42181ttcagagacc ccaaaataca gggcctcaag atgatgcctt ttttcactct cccctaacat42241tctggagaga agcagttgtt ttgcacaggt gggtcaggca ttctggttcc attttttctt42301cccgtcactg aggtgagaag aggtgaaagc ttcctttcac ctggaagtca cacggtccct42361tcagctcaca ttccaaggcc agatgccact gggtagcgtt tcataacttc caggaggtag42421tgatacaggg atgctgggac agcagcctct gccacagatg tatttttgta tgtgggattg42481aaagggagta tctgagtggc cgcctctgat atttctttca aagcattggc ccagtatcgt42541caccaccttt tctcatcaga Raagcccaag aggccgggta cggtggctca cgcttgtaat42601cccagcactt tgggaggcca aggcgggcag atcacttgag gtcaggagct caagaccagc42661ctggccaaca tggtgaaacc cagtctctac taaaaataga aaattaggcg gttgtggtgg42721cacctgcctg taatcccagc tactcaggag gctgaggccg gagaatcact tgaacccagg42781aggcggaggt tgcagtgaac catgttcttg ccactgcact ccagcctggg caacagagcg42841agactcagtc tcaaaaaaag aaaaaaaaag cccaaacgtg tggttgcctt tctgtttggt42901gagcaagacg tgagtgggct ccctggtaca aattccattt gaaatttcct cagttgttcc42961atcagctgcc aaatttccac actggcaaca cccttctgtt tcagctggga cgatgccgga43021gtgtgaagga gtttgagaag ctgaaccgca ttggagaggg tacctacggc attgtgtgtg43081agtggccaag gctaggacat gtggccgcag ctcgtggctg tgacagtgtg ggacgagact43141gtcgaagcag cagccgcgtt ggggctggaa gggactcaga ccctgtgctc tgaaacaggg43201cagaaacctg ttcaggaact tcaattcctg atgtagttat cacaaacRtt gtaccatttc43261tgggtaaacg taagcatccg gttgctaatg cccataatcc aaacgtgtca ctgcagagta43321cacagaactg ttagagaaat aaaatacaga actttccaga agcagcagga gctggacatg43381gtgctcatgt ctatcacctc agcaactctg gaggagtcac ctgagcccag gtgttttttc43441tttttgtttg tttttgagat gggtcttgct ctattgccca gactggagtg cagtggcatg43501atcatagctc actgcggcct cgacctccta ggctcaagtg atcctcccac ctcagcctcc43561tgagtatctg ggaccacaag agtacataag cacacccagc taatttttaa atttttggta43621gagatgggat ctccctatgt tgcccaggct ggtcttgaac ttctgggctc aaatgatcct43681cctgcctcgg cctcccaaag cactgggatt acaggtgtga gccactgcac ccggccaagc43741ctaggagttt gaggctgcag tgagctgtga tcacgccact gcaccccagc cttggcaaca43801gagtgagacc ctgtctcRaa aaaaccaaaa acaccacaca caactaaagg ttataacaga43861tgtgaaattg gcgtaggtca ttctggaagc tgcacacatt agcaggatgt tttgtagagc43921catcttcaca caaaataagg tttgcttttt tctgcataga gtaccttatt tgcttttagg43981acattacaag agagcacaaa gcagtgatct gcatgctaag gaggacatgg agagcctgga44041gccacagaac tgagtggctg gaggccaggg tccagcacag agcaaagttg ggcaccccct44101gcccagttca aaKgaggaaa ccctcNgggg gaacgcgtct cgggctaggg gcgttgcaca44161gagtggggac tgagtgtcac tgggcatgag gttgtcagag gaaagaacgg ggacccctgt44221ggctcaggga gagcctcccg ttcagcgcta gggagcccac gaggggcatc gagatgatgt44281catcaccaat gtgtttccat tccagatcgg gcccgggaca eccagacaga tgagattgtc44341gcactgaaga aggtgcggat ggacaaggag aaggatggtg agcaggaaat tggggtgttg44401ggacctcgca ctgggaggag gcagaaggat gtgagttacc tgaagtttcc tcagagcgac44461tgcacggtgc ttgtagcggg ggccggcctc tgggagggtt ctggtgtggc ccagcacatc44521acccccaggt ccacacgcag cctYagctgt ctgccaggag ggctctgcag tctccatccc44581ctcctccccc tcctgtctgg tgaggccacc tcactgcagg ctcagctctg ctgtcccctc44641cacccccagc acctgcctgt gcaggcaggg ccctgccatg ccccacatag atgtccacgg44701tgtttagaga ggtgccgatt cgtatttggt caaggaagag ttaaaagtgc ttattgggcc44761gggcatggtg gctcaYgcct gtaatcccag cactttggga ggccgaggca ggcggatcac44821ctgaggtcgg cagttcaaga ccagcctgac caacatggag aaaccccatc tctactaaaa44881atacaaaatt agccaggcgt ggtggcacat gcctgtaatc ccagctacta gggaggctga44941ggcaggagaa ttgcttgaac ccgggaggca gaggttgcgg tgagccgaga tggtgccatt45001gcactccagc ctgggcaacg agcgaaactc cgtctcaaaa aaaaaaaaat gcttattggg45061gtcgccccaR ttctccctga ggtggggaca ctgcagcacc tgctatcagg tgttcgtgaa45121gcccaagagt ggctggggtt ggggcttccc cgccatcact ggggtggggc tcgctgaggc45181cacctccctc cccaggcatc cccatcagca gcttgcggga gatcacgctg ctgctccgcc45241tgcgtcatcc gaacatcgtg gagctgaagg aggtggttgt ggggaaccac ctggagaggt45301acgtggtctc ctggtctgca cattgggccc tagggagcat gtgtcttggg ctagaggtgt45361tgcacagagc gaggactgag tgtcactggg catgaggttg tcagagaaga gatgacccca45421cctcaccgcc tggctcagcc cattgtctga gggggacaca ggttgtcctg cccatgtccc45481ctcctgcagc aggggaggct ccacttccac cccttaggag aaggccggag agtggcatgc45541atcttctgtt tcttccagca tcttcctggt gatgggttac tgtgagcagg acctggccag45601cctcctggag aatatgccaa cacccttctc ggaggctcag gtgcgtggca gaggggcctg45661gggtggggga atgggcttca tgggcccttg tggtgcacat ttataacgaa acagggcact45721tggggacttg aagagctgct gctgcctctg cctccacctc ccagtgtttc agcgagcttc45781agtgaggtgg aattcctgtg gtgggggcat ctgctgggag cgggcagccc cagggccgag45841agccttatga gggcactggt ttcctgggct gttgggagca ggcagccccg gggctgagag45901ccttctgagg gcactggttt cctgggctgc tgggagcggg gagccctggg cggagagcct45961tctgagggca ctggttttct gggctgctgg gagagtgcag ccccggggcc aagagccttg46021tgagggcact ggtttcctgg gctgctggga gcgtgcagcc ccggggccga gagccttctg46081agggcactgg tttcctgggc tattgggact gctagtcctg ctggccagtg tgggtgtggc46141agggcccagc tgggcttccc tgcaggctca ccctgactgg tacctctgac cctctgcaca46201ggtcaagtgc atcgtgctgc aggtgctccg gggcctccag tatctgcaca ggaactteat46261tatccacagg tgggtgacag ctaggcagag ttggaagcac aaattcggct gaggcctRac46321tctgctgccc ctgtggagtt actaaaagct caggggttcc cagctgcagc agcctgcccg46381ctgctcgcac tccatttgtg tttttttcta aaatagagaa ggggtctctc tgtgttgccc46441aggctggtct cacacacttg ggctcaagtg atccacccac cttggcctcc caaagtgctg46501ggattacagg tgggagccac cacacccagg catgcactcc gatttttaaa aggtccaaac46561atcaacagca tgtaagagtt aatgaaaaaa aaaataataa agatactacg ccgggcacag46621tggctcacgc ctgtgatccc agcactttgg gaggccgagg caggcggatc acgaggtcag46681gagatcgaga ccatccttgt taacacagtg aaaccccatc tctactaaaa atacaaaaaa46741attagccggg cgtggtggcg ggcgcctgta gtcccagcta cttgggaggc cgaggcagga46801gaatggcgtg aacccgggag gcggagcttg cagtgagccg ggattgctcc actgcactcc46861agcctgggcg acagagtgag actctgtctc aaaaaaaaaa aaaaaagata ctaaaaagcc46921caaagctgag gaaacgtgca ggaagtctac gggcattggt gccgtggggg agctctcagc46981ccctgtggcc ctctgggagc cacctgccac tgtttttcca tcacagggac ctgaaggttt47041ccaacttgct catgaccgac aagggttgtg tgaagacagg tgggtgcaac ttgggccagg47101ccctgtccct agatggcact tggtgacaca cactcccctc tctgctgcag cggatttcgg47161cctggcccgg gcctatggtg tcccagtaaa gccaatgacc cccaaggtgg tcactctctg47221gtaagtcctt ctgaagcatg gtggcccctg gggaccaggc ctgtctggtg gaggtctcct47281tggggatgtc aggccgaagc tgcaactggc cttgggaatg ttaagctaca gggtctgtgc47341acactcagaa acgttgggtc taggacagcc tccaggacac agcagggctg tgccacaaaa47401tggtgagaga cgggctcagt ggcgtcaagg gcctgcctag atcagacaca cctcgccttg47461agagctggaa acaccgtggc cgtgaaacca tctctggagg gaaggctgct tcacggactg47521aaggacaggc agcttgcatg ctttccctgg ttcttcctcc tggatacttt taaccaaaca47581gggtcatgct tagccagggc tgaatatcag agttggtcag aatcaggtca ggaccccagt47641gtcgtgtact ccccatctga gaacgggttg gtagtggctg gtgtggctgt gtgccgaggg47701tccgtggtcc ctgcgactct caggacaccg ctgcacgtga cagctctcgc ccccggcaac47761cattgctgtg tgtgccacct ggatctgcag ggcctggggc agagggtggt ccgagttggg47821acaggtttcc aggccaScac aggctctcgg ggaggctgtg tcctgtggag gcctgggctg47881ggggagagga gccggctgta ttgaggtggg tgcttctgtg tgtaggtacc gagcccctga47941actgctgttg ggaaccacca cgcagaccac cagcatcgac atgtggtgag gagataYggt48001taccgctcct ggggcctcag gaagggctgg gacaggagcc gggtcacctg gttcctgagc48061tcagcctcag gggaggggca ggagtgcagt ggggtctttg ccagcctccc actcccaggg48121aggtgggcct gagcctggaa acccaggagg aggtgtgaga acttagcttg ctgttctcag48181gctgggagca cagaggtctg gaggaggcac gcctgcccct gcacttgtca ccccgggagg48241cctgcggggc ccaggagggg agccagtggt ccctgcctgt ccttcacagt gtccctgacc48301cagSgtgcct cacactggca gggtcagcaa aggtctggct gcagtcaggt cctctgttcc48361tcgcgctggc ggggtcagca gacgtctggc cgcagtgagg tccactgttc tctgcagggc48421tgtgggctgc atactggccg agctgctggc gcacaggcct cttctccccg gcacttccga48481gatccaccag atcgacttga tcgtgcagct gctgggcacg cccagtgaga acatctggcc48541ggtgggcgtc ctgggcagac ccgcagcccc cgcccgtgcc cacgccctct gcgcccgcag48601cccccgcccg tgcccacgcc ctctgcgccc gcagcccccg cccgtgccca cgccctctgc48661gcctcagctc ctgcctccca tagggctttt ccaagctgcc actggtcggc cagtacagcc48721tccggaagca gccctacaac aacctgaagc acaagttccc atggctgtcg gaggccgggc48781tgcgcctgct gcacttcctg ttcatgtacg accctaagaa aaggtgctga tctctgcacg48841gggggcaggg accctcacca cccacactgt ccagaccgtt tcccagagcc cagcctcact48901gcggtggaga ggcccctccc caggcacagc cgctcggagt ggccacctgg cttcctttga48961gcatttgaat cacaggctgc tcagctgggt gggaggtgag gaagccgcac tcacaagtcg49021cactaacgca ggctgcctcc tccagggcga cggccgggga ctgcctggag agctcctatt49081tcaaggagaa gcccctacgt gagtgtgcag ggttcctgac tcgctctgtg gggtatggct49141gggagcccgt tttgcccggg gtgggtggtg gagaagtggc ctgctgccct cactgacggc49201acaccctttc tggggtaaga ggacaggggc atgtggaggc acagacagcc cagggccagg49261tccagaggca gagctggact cagacctggg cctgctcctc ctatctgggg ccctgccccg49321cccagcactg aacccttctc cctgcagcct gtgagccgga gctcatgccg acctttcccc49381accaccgcaa caagcgggcc gccccagcca cctccgaggg ccagagcaag cgctgtaaac49441cctgacggtg ggcctggcac acgcctgtat tcccacacca ggtcttccga tcagtggtgt49501ctgtgaaggg tgccgcgagc caggctgacc aggcgcccgg gatccagctc atccccttgg49561ctgggaacat cctccactga cttcctccca ctgtctgccc tgaacccact gctgccccca49621gaaaaaggcc gggtgacacc ggggggctcc cagcccgtgc accctggaag ggcaggtctg49681gcggctccat ccgtggctgc aggggtctca tgtggtcctc ctcgctatgt tggaaatgtg49741caaccactgc ttctYgggag gagtggtggg tgcagtcccc ccgctgtctt tgagttgtgg49801tggacgctgg cctgggatga gagggcccag aagaccttcg tatcccctct cagtcgcccg49861gggctgtccc gtgcatgggt tggctgtggg gaccccaggt gggcctggca ggactccaga49921tgaggacaag agggacaagg tatggggtgg gagccacaat tgaggatacc ccgagactac49981caggagagcc cYgggctgga ggctgagctg catccctgct ccccacatgg aggacccaac50041aggaggccgt ggctctgatg ctgagcgaag ctataggctc ttgttggata aaagcttttt50101taacagacat ggtttcacca gcgtttaatg tgctctgatg ttgaccgtcc ctctgagtgt50161tctggggagg agggggtggg gcggagggtc aggaaagcag gctcagcttc cagggtcagg50221gagttgtggg cccagagggg ctgtcacagt ggatgcaccY tgccctctcc ctcgccagac50281ccgagggtag ggcagaggca cctcctcgcc agcctgtggg ctgcacccac agggaatgga50341ggggaggggc accattacca ctggacccac caagaccccg ccgcctgggc gggctgtcca50401caggcccttc cgcccagcag cagtgacgct cagggccttc ccagctgtca gccactgccc50461gttcctgcgc ctctcgtgca cctccacatc tggtttgagg tcaggtcatt tcctgtctgt50521gggcccagct gctgtgacac ccaaggggag ggccggcgYc cccgaagcca ggtcagccgt50581gcacccggca gagccccgca gattggcYcc agcagagctt ggggtggggg gagctcggtg50641tcaccaacag gcccttgagg acactcgtgt ggagaatccc tgggacacgt ggaggacccc50701caaStcctga gccccgtact ccgtactgca gggagcaggc caggagccac gggccttggg50761gcacagggtc cttctcaggg acaggttcag gcactggctg gaacaggctg gacccctcta50821cccagcacat gtgggcatgc gtctgggttc cgagggtggg gaaatgtgga aaggctcctg50881ctggccggct taggcctgct gcccttggag ccttccatat acagagagtc ccacctccag50941caagggttgg cctggactct ccaWccccct gctgtgccca ggactccccc agggacaagg51001caaccagagg cccagacccc tccccagcaa agagaagcac cacgggagct gtctcccaag51061agcccttgac ctggggccca gctggcctag ggccgggccc cggggctgtt gtagagcaga51121gtgtccatct gtgcacgctg tagccacaag cgccgctggg cgtcgccaag cagcacacgc51181agcgagtggc ctggacggca ggcgggcagg gctgcacagt gggcagcatg cagctggcga51241caggtgtcgc agcagagggt gggcagggcg ccaggggaca ggcagctgtg tgcctggtag51301cctgggggct ccgagacaga cctataggct gaggctggta ccccctctgc ccgaatacgc51361ctcgggctgg ccgctgcaga gctttcaggg gtggccaggt ccccaggcct actcagctca51421cggcgcagag agaggaagga gaaggcggaa ggttcaggtt ccagcccctc ctccaaggcc51481ccatatggtg ggctgctggc ctgcggcagc tcctcagctg ggggctccca ggcccggccc51541ccagtgcccc acagtttggc actctgctcc cagagtggtg gcctgtaggc cagctccgcc51601gggggcgagt cagggcctgg tgacggctcc ccatacaggc tggcgtcctc cgacccctcg51661tcttcctgca agtcccggta taagtccagt ggggccctgt aagcagcagg ggagcctcgg51721gggggcaggg gtggcggctc ctcatcctgg gccagccgct gctgcagcca ggccacacag51781gaggccacgt ccccgctggc acgccgtgcc tgcagcagct cctcagctgg cagcacactg51841gtgcccagct gggccagcac ctcacccagg atctcacact ccagccggag ggcgaagcag51901cccagggcca cctgcaccag ctggcaggcg gggggcaggg cggtcaccat gagccgatgg51961ctgtctctgc gtacgtagcc catcttctgg aagctcttca ggaggaggtc gtctgagagc52021acacccttca gcacgtgcac gtagccccca gagaaggtct gcaagggagc ggccaggtca52081gtgactgccc accctcccat aggccagacc ctagggtaca gtgggtccgg gatctgttgg52141ctcaggcctt ggcagcttct gatgctcaga cccaacccca gagactaggc ttagccagct52201tgggccaggc ccaaacatga tgatttcttt tttttttttt tttttttttt ttttttgaga52261cagagtcttg ctctgtcatc caggctggag tgcagtggct ccatctcagc tcactgcagc52321ctctgcgtcc caggttcaag caagttcccc tgcctcagcc tcccgggtag ctgggactac52381aggcgcccac caccacaccc ggctaatttt catattttta gtagagacag ggtttcacca52441tgttggccat gctggtctca aactcctgac ctcaagtgat ccgcctgcct tgagctccca52501aagtgctggg attacaagcg tgagccactg tgcccggcct aRaatttttt tttttttttc52561ttttttgaga cagagtctcg ctctgtctgg agtacagtgg cacgatctcg gctcactaca52621acctcctcct cccgggttca agcgatcttc ctgcctcagc ctcctgagta gctgggatta52681caggtgcgca ccaccacgcc ccactaattt ttgtatattt agtagagacg gggtttcacc52741atgttgacca ggaaggtctt gatcccttga cctcatgatc cacccacctt ggcctcccga52801agtgctggga ttacaggtgt gagccaccgc acctggccga atttcttaaa tttaatggaa52861aaaacaatga atacattaac ctagctcaca aatcagaaca aaatgaagag cccctgtact52921gagaaattcc actgtcttct gtcccaccgc tcccttcacc cctggagatg gcctcatcta52981attagttttg aatgtacttg cccaggtgtt tgttttgcaa acccaaatac agatttcgtt53041tgctcctcgt atgaggtagt cccccctttt ttttcacctc tttgtaaggt ctctaaccaa53101agctagacag gtagcctctt aaacgtgttg ttctgaactt tgtttttttc cactacagtc53161ttgcagacaa ggacagaagc tgtaacgttc attccccctg ctgtgtagta ttccaccatg53221ggggtgacac gtctgttttc agcgctctcc agaggctgtc catgcactgg gctgacagcc53281gtgggacggt agaagggtga gtgctcNttt tttttttttt ttgagacgga gtctcactct53341gtcaccagga tggagtgcag tggctcgatc tctgctcact acaacctcca cctcccaggt53401tcaagtgatt ctcctccctc agcctcctga gtagctggga ttacaggcgc gtgccaccac53461acccagctca tttttgtatt ttttaataga gaYggggttt caccatgttg gttaggctgg53521tgtcaatctc ctgacctcgt gatctgccca cctcggcctc ccaaagagct gggattacag53581gcgtgagcca ccgcgcccgg cctggctgag tgctttctag cctacactta ggtgaagaac53641taaaggagct caggacaatc actgcgagcg tctcctgact ctcaaattcc ctaactccgg53701gactcctagc atctaatgcc tggacacggc ccacgtgtgg agcgcccagt gcctgcagca53761ctgagctcgg ggctttgatt cattcctctg gccccatgaa ctggactgtt cccgtgtcgc53821tgtctcagga ggaaaggggc tgacagagat tagctaattt ggccaagtcc tcgcggccag53881gaagcagcag aactaagtcc aaaccagggc tgtccSgcac tgaagccggt tattcccccg53941ccccgccccc tgggaggcct caagggcggg cctgtccgtc ctctctcatg aatgaaagtc54001ctggcgcctc cgaccagcag gtgtccccgg ggagccgcgg agagactgcc gctgccatgg54061caaccggcgg aggcgtagct cgcccacgtg cccagtgacg tcatcggctt cctactcttc54121cgaattagct cagagctccc ggcggccctg ccccctcctg ccgccaggag ctcacctgga54181gaggccaggc cagccctggg cacagcctcc cgccccaggc cgtggagttc actaggtctg54241agcccagccc tgagacttca cagggatgca cccatctgca gtcctcccag cctcggcggc54301ctcctctgca aatgggattg gcccataacc tggaggaatt aggatgacca tgggaactca54361agtcccttct gtgcccccgt cctaagattt ccatggcaac tggggccgcg aggcctctta54421gggaagacac tgacgcaagc tctcagggca ggaccccgta ggcagcaccc aagcctgccc54481tcgcccaccc catgttctca gccctaggag tgggtgcccc cacacgccca cgacccctca54541gcgctcggag ccttgaggag cccaccaggc actgaggaca cttgaagccc ctctctctcg54601aaaggcgtgg tcccccatgg tcggcggccc cgcgacccgc aggccaggac gaggggcgcg54661ctcctcctgg cctggcgcct accttgatgg tggtgaactc cttcctccag ggcagcaggt54721acaggtgcac cgcggcgagc tccagaagct cgaaggcgcg agccaggccg cgtagcgcgg54781gcgccaggtc ggcgcggccc cacagcccgt cggtgagcag agccagcgcg tcgtcctgca54841gcgccccgtg caggtcgaag tcctccacca ggatctgcca gagcaccgcg cgcagcgagg54901ggtccccgca cacgcccgcg cggccccgtc gcagctcgcg ctccaggcac tggcggtagt54961cctcggacag cgagctgctg cccatcctgc ggcggacggg ggtcgggggg gtcagggacc55021taaggccgcg cccagcccgc cgcgctctcc ccgaaccctc acccagcgtc cccaggaccc55081ctcgagcctg gagaccccgc ccccagcggc tgggcgcccc cggtatcttc ggcaccccct55141acgctggaga cctcggcccc atcagctgcc cgcccaccca gcgatccagg cccctccagt55201tgagcgcccc cgcacctcca gcccctccct ccagcctggg tacctcggcc ttccaggggc55261gcccccggtt atcgaccgca cccttctcta gagacgcgct tccgcccgct gagccctctc55321ccgaccagag ccccttcgcc cggcagatgg agctgcggat cccgcgggtg acggcgctac55381ctgtctgtcc ccagcggtct ccggccgccg cggatcagtt cgggcctccc cgcgcgccgc55441gcgcttccgc cgggagtcgg ctccgcccgg ggtccgcggc cgtgacgctg ggggcggggc55501ggggcttggc gcaggccccg cctccgaata atttaggagg ggcggggcct ggtgaaaggg55561gaggaggagg tgtcagcccg gccgagcccc tgcgcgaggc accccgagag cctcctgtca55621ccgcgtcgcc cgtctcctcg cgggaccggt atcgttcacg acgccgctgg gccgagggct55681gcgggctcat ggcaggcgct cagggaacgt tgtgcctggg ccgctcctcc ccacgagaca55741gggacccagc cccaggcgac ccggccgcgg ggaagcaggt gcgcgggctc gtcccctggg55801gggtggggcc accggcatta cctggcgggg attggctgga tcctgggaag agcctttcct55861gcggggcggg gccacctgca ttacctggcg gggattggcc ggatccgggg gcgggcctcc55921cctgtggggc ggggccatcc ccttgaggga ggggcccgcg gcacggaagg tgggcggaag55981gacaagcctt tcagcctcgt ctgaaacagc agcctcaggt ggatgcgggg acctcagatt56041ctgtttatgc tggctccgct tcccggcctc ctgcacttgc catctgcagc ccttgagagc56101atcgtctggg cgggtctctg aggcagggcg aagccgcccc cctctctagg ctaaaagctg56161acctgggctg cggatccacc caggctaaat ctgggccacg acttcccggg gacgttttct56221tcgctcgggt tcctctgttc tgtgctacag ccactttctc gcattgccca gaagtagagg56281aggatgaggg agacccttgg aggtcccagc cgaaacccac ccctacccca cacccccgct56341ttgggcaggc tttgctactt cacgtctgtg tccctcatct ctccaggcca ccaaaaccaa56401ggcccaagtt tgttgaaact ttccctgggc ctgggcggcg aggtcccccc tttttcttag56461attttttacc tggaagttcc ctgctggcat cttagaaata gtgatcatga ccgggtgcgg56521tggctcacgc ctgtaatccc agcactttga gaggccgagg cgggcggatc acctgaagtc56581aggagttcaa gaccagcctg gccaacatgg tgaaaccccg tctctattaa aaatacgaaa56641gttagccggg cgtggtggca catgtctgta agcccagcta cttgggaggc tgaggcagga56701gaattacttg aacctgggag gcagaggttg cggttgcagt gagctaagat cgtgccactg56761cactccagcc tgggtgacag agcgagaatg tctggaaaaa aaaatcgtga tgcagtcggt56821ggatctccct gagttatcta ggtggccacc tctggacagg gagctctttt acttactgtt56881tactgaactc actcttggtg cccagtgcca ccatgggtcc acaccagtcc tcaggacagc56941taacttcgta agtctcatct ttattcttct tctgatgcgg aggagggtgg aggtaactgt57001tctgcctttg tcctcactga atttggcaac accagtccat ccatccagca gtgggcctag57061ctctcctccc ataggcctgg gtgggttttt gcacctgcct caacccatgg aatagaactg57121atgtggaatg aattccaagg ctaaatcata aacatgccat accctggcct tgtcttgaga57181tccttggcct ctgacatggt ttggttctgt atccacaccc aaaatctcat cttgaattgt57241aatcccacag gtctggagca ggaccaggtg gagataatta aatcttgggg gcagcttcct57301ccatgctgtt ctcgtgatag tgaattccac gagatccgat ggttttataa ggggttctcc57361cctttgttca gtgctcattc tcgctcctgt cgccctgtga ggagacgcct tctgccatga57421ttgtgagttt gctgaggcct tccccagcca tgcggaactg tgagtcaatt aaacgtcttt57481tctttataaa ttatccagcc tcaggcagct atttacagca gcatggggac ggactaacac57541accctcagaa cccagccact agagtgagaa ggccacggcc gtgtgcagag cttttggcct57601gcaacttcga cctgcgactc acattcacct gcagcctatt cttgggactt ttttggaaat57661agtttttcca gacaaaaatc aagatgagtt tgttctgggc cgagcgcagt ggctcacgcc57721tgtaatccca gcactctgga agaccgaggc aggtggatca cgaactcagg agatcgagac57781catcctggct aacacactga aaccccatct ctactaaaaa atacaaaaaa ttagctgggc57841gtggtggcgg gcatctgtag tcccagctac tcgggagact gaggcaggag aatggcgtca57901acccaggagg cggagcttgc agtgagccgg gatcgcgcca ctgcactcca gcctgggcca57961cagagcaaga ctctatctca aaaaaaaaaa aaaaaaaagg atgaggttgt tctgggttgg58021catgggccct caatccagta tgactgctgt ccttatgagg agagagaaac ttagatatga58081acgcacagga gagaagaccg cttaaagacg gaggcagagt ggaacgaggg tgccgtgagc58141gaaggtaaca cagctgtgtt agagccacca gaaactggag gcaaacagaa ttctacccta58201aagcctccag agggagtcag ccctgctgac agcttgattt tgacttcagg cctgcaagat58261ggtgagagag aacatttctg ttgctgtaag ccacccattt tgtggtcgtt ttggcagccc58321ctggcaacac atccagtcct cacgtcgtcg ttccgacagc cctgaatgag gtcccagccg58381acagctggca ccaaccatag acagcagaca gctcttcgga tgaatccaac ccctggtcat58441tgggttacca cctagaaaca gcctcccatg atgctgagga gccatctggc tgatttttgt58501actttttgta ttttcagcag agacagggct tcactatgtt ggccaggctg gtctcaaact58561cctgacctca ggtgatccac ctacctcggc ctcccacagt gctgggattg caggcgtgag58621ccacggtgcc cagcctaatt tttaactttt tagtagagag gggggcctta ctatgttgcc58681caggctggtc tcaaactcct gggctcaagt gatccccccg cctcagcctt ccgaagtgct58741gggattccag gtgtgaacca ccatgcctgg cgtcttgagt atttaagccc ctgagttggg58801gtagtttgtt aggcagcggt ggctacgtgg gctgtacaca ttcacggctc cctcccttcc58861tgcccgggga tggacactcc atgagctgcc gtgtggattc tccgtccttc cctgccctcc58921cgggacagac actccattgc ctccactgca tatacatcct ctaaaaccgg cagcagcctt58981gccaggtcgg aggttccgtc cacacttacc tggactccag gactgcacct ggtcccctgg59041ggccactgcc gtgtccaccc cacagtgtgc gtgagctccc tccccctcct gtactcagca59101tcgtccagct tcctcatctt ctccggtcca atggatataa ggttacatct tgtttacttt59161tcatttgatt tttgttatga tttggacact gcttatgcag aatagtttta tttttatttt59221tttttagatg gagtctcgct ctgtgctcag gctggagtac aatggcgtga tctcggctca59281ctgcagcctc tgcctctcag gttcaaacaa ttctccggcc tcagcctcct gagtaactgg59341gactaccgcc accacgctca gctaatttat gtatttttag tagagacggg gtttcgccat59401gttggccggg ctggtctcga actcctgacc tcaaaagagc cgtccgcctc ggcttcccaa59461accgctgggg ttacaggcat gagccactgc gcccagccat tactttttat tttattttta59521ttttttgaga cggagtttct ctctgtcgtc caggctggag tgcagtggca cgatctcagc59581tcactgcaac ctctgcctct caggttcacg ccattctccc gcctcagcct cccgagtagc59641tgggactaca ggtgcgggcc accacgcccg gctgattttt tgtattttta gtagagacgg59701ggtttcaccg tgttRgccag gatggtctYg atctcctgac ctcgtgatcY gcccacctYg59761Rcctcccaaa gtgctgggat tacaggcgtg agccaccgcg cccggccatt tgtaggattt59821ctaaaaggaa gctgctatta ttcattcctg tcttatgggg gagctgcagg ctcagcgagg59881gcaaagctgc acagcaaaca gcagcacttg aacatgtctg ccgggtctca gcgccaggga59941tacagctgag ccaggacgtt cccRgccccc acagtgtgca ggctgcaggt gtggccccga60001tgccaagcca gcaggggctc ccacgccagg atttgttcaa gacacagctc atctccacca60061ttctatgaag cggactttaa aatcccatct tctagacacc aacgctgagg ccggagcagc60121ccaagctcca aggcaccgtg gccagctggg gttcgtatcg agggctgggg cagatgctcc60181agctctgccc cacccagaga gacacttctg tctccctatg gggtccttgg cgggtaaacc60241agcccctgct Sagggtgcaa gcacagagca tcccaccgag gtccacagct actgctttta60301agactgtttc cctgagaaca gcaacactgc cccccagcca gctcctggta gcatctaggg60361ttgggacctc ccaccccacc agccctgcca cctgcctcct catggcatct caaggtgcca60421ggggtgcctt tcttgcagaa gagccctcca tcctgatctg cccacaccgc cctggcagat60481gctgtccacg cccggcagcc ttctgtgcac ctcagagcat gtgtgtgtcg ccatgcacag60541aacgatgatt cctgagaccc tcgggtgact gctctgggct caggtcccca gtgatgacct60601ggaccccagc ctcaccccac ccttcccacc cactgatgcc ctgcagagct ctcctgtcca60661agagggtgaa gcccacacct gcctggccag gcacagcccc aagaggaccc tgcttctatg60721cagagggtct cggaggctgg cagatgttca agaggttggg ccaggcttgg tttggggctg60781ggctgtggct cgtcccccac atcctcagcg gcctctgcca tgggcagcag gggtggttgt60841gctgcctggc cagtggggtg gggattgagg gagccacacg gcgaagcaca gcagtgagct60901ctttatttta ggctgggtgg gggcagcatg tgacagtcaa gagggctgac ccttgggctg60961cacggaccta gataagatgc ccagggtccc ggaatgttca cagcccacag ccaaggtgag61021taaggggcag ggaatgtcct gggctcagag cccgagaccc tgcaggggtg tccctgccca61081tcagcaaaca gcataccctg acaccagctg caccccaggc cccagccggc ccttgttcct61141gacccagaca agaaactagt tcatggggct acaactcccc ttctaggggt cagaggggag61201agaaactgga ctgccgtctc cactgctggc cggctcatgg ccccctcttc cctgatgctc61261cccaggagct gccccagcca gatgttcatc gctgagcctg cacccaccga agcccaaagc61321ttcccttctt ggcctctctg aggcctacac aggagagcag ggcctggttc ctcctggagc61381tgattcagcc ccattctgtc ggggcagagg tgccaacccc aagctccagg gtcagatgtg61441aggtgaggaa ggtgaagagc tggagagcac accacagtgg acaagccccc accatgtgca61501gagcagcgtg aggctccagg atggtcctca gtagatccca tggctccagg tgtaggagag61561acagccctgg gaggcgaggc caggccctgc agggaccctg ggctctagct gtactacttg61621gccaggcctc cccggggcag cagctccacg tagctcaggg cactctgcag tgatgtgagg61681cagtagccct cctctccgat caggtacctg catggatggc aggggcagcc ggtgtcatct61741Raacgaacag aaggataccc ccggccctcc agaaggtagt gtctgccctc ccagttcagg61801acctgggtgc tgagggtggg caggtggggc tgtcagggcg gctgggctag aggaggcctt61861tcccacccac tatgtgctcg tccttggacg gtagagccag ggatggcact ggatccttat61921cccctcccca ggcagatgct gctcagggga cagcccccag ctcctgctgt ctgggagagg61981ctgagctaag cgaagggcca cgggacatgt ggcagcaggc actctgcaca ggtggaccca62041caggggctgc ccgaaggcat ggctgtgacg gagccacacg ctaccctccc cagctgctac62101tgacaaccac cgtttaggtt gagggatgag gtccctgctc ttcgcacaga gacccccacc62161ctgacccgcc agggtctacc cctcgtggat gaactcctcc agggccgcgc actccgacac62221cagctgaggg aggccgctcc tcagcaccac gaaggacagg atgggcagca ggtcatcggc62281accactgtcg ggagggcata gcggccttgg gttggggcca ggagcctctc actccgcaaa62341cccagcagct gtggagccat cctggccgtg ctggccgcca tctctgtgcc cttctgcgca62401gaggcaccgg aaggcagctg ggcatggtga ggctgtccct aaagggagct cccgtactgc62461ggagccaggg ctgggcccac acggccctcc tgcttctaca gtgcagccag ctggcacaag62521gctaaggggc aggaagatag tccctcccgt gaccattgcc tttctagggc ccttcctccc62581tggccacacc tcgccctcct gggactcaca tggcagctgc agcgatgggc gggggcccgg62641cctggggtgt ggcctctggg gtggggcagt agtcttccgc acagacacag atgatccgca62701gggtccgcac tgcgccccag acagggtttc aggggcagtt aaccccgtcc agcagcctga62761gccacccctt ataccccgcc cacccaccca cctcctgatg tgcacggctg ggaagagcca62821cggtgaccct ggcgatgttg ctcagacacc acttttgttc tggtccccca acctccccac62881cggcacgtga caagagaagc tcagtggtga agacacaggc tccaggtcac ctactgggat62941gggagcctgt ccctccccct gactctcgaa cctgctgacc tatgcactcc agcttcttct63001gggggcagct ctccaggacc agcagtccga gctcctgggc cgccgcgcag taggggtagc63061cagtggcccc cttggcctca gggttctggg ggaggagctt ggtggggatg ccaatggcgg63121tggggggtgc attcctgtag agctccatgc tcctgctcag ggcagcctcc cgggctcggt63181gcacgctcct ggggcagaga gaggccaggc taagctgccc cagggacctt gccctgggga63241cagaagcagg gctgtggctg gagctcagtc ctggagaaaa ctggctatgt atctcaagca63301aaaagcccca gcgttgccaa gaccccaggc acgggagagt ggcaataatc ctacctgcaa63361actggaagcc cctgaggatg gcctcagggt aaagcacagg ctagggggag caaacacgag63421tcttggggat ggaggggagc agacagtgcc agggagcagg gcaggcctcc tgggggactg63481ggctggaaga cactaggcca cgcaggtggt tgtacagctg ggtccatacc tgtacagggc63541cagcagcaga ggccacagcg gggagaaaaa gggttcctca atgcaggcca ggcagcggtc63601cttggaggca gctgtgttta ggccttcgaa ggccagaagg gtcagcgaga gcagcctgtc63661tgctaggaac agagccgggg acgtccacag aggctcctcc cctgctgggt caagtggctc63721taagccacgg ggcctttctc cgcccaagaa acctctttcc accgccttca agcctcctgc63781acccacccct gaccctcctc ggcccagggc agggagtccg ggactggaaa cggtcaccta63841cagctgctgc cctgttgggc cagcagcctg agaagaaacc ggagaatgat ccttttctca63901aaaggggctt tacccaaaac tgcaggaacc agggaagact ttagatccaa ccccctgtgg63961gtggggaccc tgaccaagtt ccttagtgtg tgggcagcac agtgagaatg acgcctggac64021tgttgggcct ccctcagtgg ttcttagaaa actctgggct gggcgcggtg gctcatgctt64081gtaatcccag cactttggga ggccggggtg ggcagatcaa ctgaggtcag gagtttgaga64141acagcttgac caacatggtg aaactccgtt tctaccacac tgtttccaaa atgagaaggg64201ctgaggcgtg agctgggtgg atgtaagtct gatcagagtc tctgatcaac agacccttcc64261ccaaacagaa gcttgaaacg gaccaataag caggggtgaa gagctgcggc ggctctccca64321gggagggcct ggcctcacct acggcattgt ggatctcctc caccgcggtc ttcagctgct64381gcagctgggg ctccggctgt accccacggc cccggccctg ccgctcagac gtccccagga64441actgctccag gtcctcgaac gagctgtcct tgtctggcaa tccagatgcg gtgtccccca64501ggggtgaggg agaccccacg gggccgggtt ggagtggggg cgtgggggga ctgtcctggg64561gccgcggggc tgcgctgggc tcgggcgggg acagcatgca atggaggctc tgcgagggcc64621gcagccgtcg gcttccgggg ttgggggtcg gggggcagca gcctggggct ggcgcggctg64681ctctgctcac ggcgggatac agggcgctgt acacggagca ctgcagccgc ctcaggagct64741gcgcgatcgg gtggtcgggg aggctgcggg agaagaggac gtgagaacgc ggccagcctc64801ggcctctccg tggcatccac aggtgccgcg ctatgcccgg tttcagagaa gcgacttgca64861ttacctcctg acagtgaatg ttggaaagtc tgccccccgc ccccgctggg gaggttggag64921aagccgagga aacgccctca gcgcaaaggg agctgcagtg tccacgtggc cgggcacagg64981ctccctgtgt cagtccagcc cctctccggc acctggcacc aggcagaggt gcccaaggcc65041ggtgcgcagg ggctctgtgg ggcctgaagg cccagctacc tgagcaggtg tgagaccagg65101ctggtcacga gtgacaggtc ccccggattc ctcttgagct tggccttcca gtgcttcggc65161cagtcctacg ggacaggggg ccttgaagga aagcacggtg ggtccgccac gaccggacgc65221gcctcatctc ccaccagtgc cgacccctag agactgtggg agcccgtcct ggggtgcagg65281agacccagcg ccagcaggcc ccgcgacccg cccccgcccc cacccacacc cgagggaaac65341cttaggcgaa ccctgggcct gcaccctcgc tgggaccctc cctcctcgca gggaccctcc65401ctcctcgctg ggaccctccc acgcgacccc tgacctcgcc tcctcgctct gccccgcccc65461acgcagccct ggccccgccc cctcaccggg ccctcccacg ggcctctggc cccgccccct65521cactgccccc cacagcctct ggctccgccc cccacggggg cccacccagg cggctctgac65581cccgcccccc tcgctgggct cctcctacaa agcctgggcc cgtgggggac gcacatggtc65641ctgttcgtac tccaggatgg cggcgtaaag ggcccgctgc tcccgttcct ccggggtcag65701ggcgacttgg ctgcaaaacc tcctggggag aaaggcacgc ggtgggtgcc ggggccaggc65761cctccccgca gcggccccag gggcgggcag gagtcccacc tgttggcggc ctcctggagc65821cgcagccggc gctcctccat ctttctctgg agctgggtgc agagtcaagg ggccgagcgt65881gggatctgag cgggccctcg gttcccaacc ccgtcccaca cccggagagg ccgggacgtc65941ccagggacct cctctcggtg aggggctgtc cccaggtaag gcttctgggg cagaaggcag66001cctgcgctca agccaggagg gccgggccag agaacatggg ggacggcggc cgggccttcc66061cctgagaccc tgcctctgca aagcggggag cccctcggcc ctgtaccccg accccggaac66121ctccgctccc ctcgctgccc ccttctctgc ccttggctca aggccacact gagtttcccc66181tcggggacca ccagggcggt cagaaaggct gctgtctcct cccgggcttt ggcaatcact66241aggttctcca tcatctgccg ctgtagagag agggtctggg gggggaggag ggagtaaggc66301cgactcccct ctgccccgcg cccacgcacc cccaggccct ccctagcccc cactcaccag66361ggatgtcttc tgcatggcct ggctggggtc tagccgcgcc attcgggcct catacgcagc66421cttcagcttc tgattctgca gggaggcctc ctccagtggc gtcagctctc tggaaatgtg66481acaagctgct gtctagaccc caggaagccc agtggtgtct ggacaccaga ggagtctctc66541tcatcctccc ggattgctct gaggccagga ggtgaggaga aagaggaggg gagccatctg66601gggtgggctg caacatctgc cccttcttgc ctgcccttcc aaaacacagg ttttggccag66661gtgcagtggc tcccgcctct aatgccagca ctttaggagg ctaaggccgg gcggatcacc66721tgaggtcagg agttcgagac cagcctggcc aacatggtga aactctgtct ctactaaaaa66781tacaaaaatt agctgggcat ggtgcacact tgtaatccca gctactcggg aggctgaggc66841agaatagctt gatcccggga ggtgatgcag tgagccgaga tctggccact gcactccagc66901tggggtgata gagtgagact ctgtctcaaa aaaacaaaac caaaacaaaa aataacaaaa66961agaaagaaaa cacaggtttt ggtattgcca ggacttttgg tttttctttt tttctttttt67021tttttgagac ggagtctcgc tctgtcgccc aggctggagc gcagtggcgc aatctcggct67081cactgcaaac tccgcctccc gggttcatgc cattctcctg cctcagcctc ctgagtagct67141gggactacag gtgcccgcca ctatgcccgg ctaatttttt gtatttttag tagagacggg67201gtttcaccgc attaaccagg atggtcttga tctcctgacc tcgtgatcca cccgcctcgg67261cctcccaaag tactgggatt acaggcgtga gctaccgcgc ccggccggac ttttggtttt67321tcaagagaag ctggaccact ggacttctat ggagatccgt tatatgtgtt tggcaacagt67381atgaatcttg ttaaagcaag cttgtgcaac aatccgaggc tgtatgactt cgaatgtggc67441ccaacacaaa ttcctaaact ttcttaaaac atcactgagg gctgggcacg gtggctcacg67501cctgtaatcc cagcactttg ggaggatgag gcaggcggat aatgaggtca ggagattgag67561accatcctgg ctaacatggt gaaaccccgt ctctattaaa aatacaaaaa attagccggg67621tgtggtggca tgtgcctgtg ctccagcctg ggtgatagag actctgtctc aaaaaatcac67681tgaggttttt ttgcaaaatt tttttttttg agtcagtctc attctgtcgc ccaggctgga67741gtgcagtgac acaatctcaa ctcactgcaa cctccgcctc ctgggttcaa gtgattctcc67801tgcctcagcc tcccgagtag ctgggattac aggcatgtga caccacgccc agctaatttt67861tgtatttttt agtagagatg gggtttcatc atgttggcca ggctgatctc gaactcctga67921cctcaggtaa tctgcccgcc ttggccttcc aaagtgctgg gattacaggc gggagccact67981gcgcccagcc ttgcaatttt tttttttaag ctcatcggct gttgttagtg ttagcgtatt68041ttatgtgtgg cccaagacaa ttcttccagt gtggcccagg gaaggcaaaa gattggacac68101ccctgcttta aaggcactat gtgggtcaaa cagccagatt agtctgtgac ccatgaaagg68161aagaagtgtc ctgtctcctt tatacagttc ccaacagcca ctccaccttg aaagctttgt68221tccctcgtac cttcccactg tcagggacaa tggaggacaa ggaggaaagt cagagcctga68281cgcctggcca ccctgccctg aagccacatt gcagaaacaa ggggactggg aagccaggcc68341ccgccagagc aggggcagga ggagtggacc cttctcgtct ctcccacttg ggagtcactg68401ggctttttgc atttgtgtct ttgtatcttt catcagttct agacaactgt ctgccattct68461aacttctaat aatgcctctc tcccattttc tttcttcttt ttttttgagc cggagtctcg68521ctctgttgct aggctggagt gcagtggcac gatcttggct cactgcaacc tccacctcct68581gggttcaagc gattctcctg cctcagcctc ccaagtagct ggggttacag gcacgcgcca68641ccatgcccag ctaagttttg tatttttagt agagatgggg tttcaccatg ttagccaggc68701tggtcttgaa ctcctgacct tgtgatctgc ctgccttggc ctccctccca ttttctttct68761ggaactccaa cgaacacgtt aggctttcca tctcttgttt tctgtcttta ttttttgttt68821ctgtaattca ttctagctat cttttgacct tttttccaat acatgaattt gctcttcagc68881tatatctgct ctgctgttat aaacttgtct gctcagtttt gtatgtatgg atttctacat68941atatatagat atataactca tccttgttcc cagctaaatt tttttttttt tttttttgag69001atggagtctt gctctgtcgc ccaggctgga atgcagtggc gcaatctcaa ctcactgcaa69061cctctgcctc ctgggttcaa gggattctcc tgcctcagcc tcctgagtag ctgggactat69121aggtgcccgc caccacgccc agctaatttt ttgtattttt agtagagatg gggtttcacc69181ttgttggcca ggatggtctc gatctcctga cctcatgatc cacccacctc ggcctcccaa69241agtgctggga ttacaggcgt gagccaccgc gcccggtcat tttttttttt tttagacaga69301gtctcatttt gttgcccaag ctggagtgca ctggcacaat cacggctcac tgcaccctca69361acctgtcagg ctcaagtgag cctcccacct cagcctccct agtagttggg actacagggg69421cacaccacta tgcctaattt tttttgagac ggagtttcac tcttgttgcc caggctggag69481tgcagtggca caatcttcgc tcactgcaac ctccaccttc caggatcaag cgattctcct69541ggactcagcc tcctgagtag ctgggattac aggtgcacac caccacggcc agctaatttt69601tgtattttta gtaaagacgg ggttttacca tgttggccag gctggtcagg aactcctgac69661ctcaggtgat ctgctcgcct cggcctccta cagtgctggg attacaggcg tgagccacca69721ggccgcctgc atttgcttgt gtccggccag gggatattcc cattctggga ctcattttaa69781aaacattcgc tgtcaggcca ggcgcagtgg ctcatgcctg taatcccagc actgtggggg69841ccgggcgcgg tgactcacgc tgtaatccca gcactgtggg aggccgaggc aggcgcatca69901cgaggtcagg agatcgaaac catcctggct aacatggcaa aaccccgtct ctactaaaaa69961tacaaaaaat tagctgggtg tggtggtggg cgcctgtagt ctcagctact tgggaggctg70021aggcagaaga atggtgtgaa cccgggaggt ggaggttgcc gtgagccaag attgagccgc70081tgcactccag cctgggtgac agagtgagac tctgtctcaa aaaacaaaaa aaaaatcgct70141gtcatcagct catctttctc agcctcaaaa acccaatccc aggctcatcc cacctcccat70201cttgtccatc ttacttctta cagctttgtg actctgcccc ctgaagcttc tggaagatct70261cgggtggcag aaaaggagag agctttcctc cttcatcgga gtatacacgg cggtgtcggc70321cggcaggctg gggaatggga gcagctgcag gcatggttgg cttcaggcgt gttttccctg70381caagccatgg gtaaccaggg gtcaagcggt gggccacatc ccacagagga gatgcgggct70441tgggggatgc cccaggagag ccgcctactg caggagaccc accaagcttg gcggccgtcg70501actgggccct ctccagacac tgctgtgcta gcttcagcat cttggaggtg tcggggggca70561cagtttcccc agcttctggg ggagggacaa gaaggctggt cacagtcggc tctaccactg70621ttacttactg ctgagataaa atgaggcaac ggaaaaggca ggcaagccca ctgcccccct70681cagcaccccc actgaccccg ggcaagccca ctgccccccc atcccctcac tgaccctggg70741caagcccact gcccccccat cccctcaccc cgggtaagcc cactgcccct cccgtccccc70801aactgactga ccctaggcaa gcccactgcc cccccatccc ctcaccccgg gtaagcccac70861tgccccttcc accccccact gaccccgggc aagccgacta ttcctcatgg accccaggca70921aggcccctgc ccccaccatg gaccccaggc aagtcccccc acccacgcat ttctaatcat70981ctgccctggt tttgcctcct gagtctgcta aggctgtggg cccctcatcg aggcccgtca71041cagcctcgac cctaaacagg cctttagtgc tcactcagct tcacttgcaa ctgcaggcct71101ctgggaagtg gccgttttac agagacaagg acgtgctgag gggatcgcct cacccatctc71161ccaccggctg tgtccccaaa gtcctgcctt cccgtagctt ggcatggagc acctctgcgc71221gaccatggac ctgctcacac agcccttctg cgcgccgtgg gagctgcctc tctgtgactc71281gcccctggac ccaccgcccc agcgcctgac ccacctccct gctgcccttc acagaaaact71341tctcatagac tcccattttc tcatcccgcc ccgatttagc ccccgactcc ccccggaaac71401tgcccttacc cgtccccacc ttgccagaca cggtgggcca ctggagttcc tctgtcttct71461gggactttgc tctccccagg ggatcctctg ccactcctcc acccaactcc tcaatcttgg71521agcgccagag ctcagccgag ctcaccggct ccctagggag ctggggcaac tgttagctcc71581ccctgacctg tgtgatccta cgtccagtgg ctgccccgac ctctgtgatc ccacgtccag71641gggctccccc gacctctgtg atcccacgtc cagtggctcc cccgacctct gtgctcccac71701gtccagtggc tcccccgacc tctgtgctcc cacgtccagt ggctcccccg acctctgtgc71761tcccacgtcc agtgggtccc cctgacctct gtgatcccac gtccagtggc tccccctgac71821ctctgtgctc tatgtccagt ggctccccct gacctctgtg atcctttgtg cagcggctgc71881tggcctctca aacttaatgt gttcattgcg gaacttgcta ttcctctcct tgccagtttg71941ctcctgcccc atctgcctcc tgcaagtctg ctctgttcca ccttcaggac gcaccccgag72001gccgtttact gctcccatcc tgggtggtct ggccactgcc atcctcagtc ctcacctgag72061gctccactcc caggctgtcc tctccctcaa agctcaaagt ctcttcaagt ttccctgcac72121accagaacgg ccccagttcc acccaaggca tgcacacagc cccaccccgc ccactcctat72181ttcagggcct ttgcacaccc cacagctgga atgctgcccc acgtctgtgc cgctcactgg72241agccagtgtc ttgccacacc ccagtctcca tcttcttatc cacaacgctg gtcccttcct72301gccactatat cacactgagt gtgggctggt ccacctacta caaatggtca ggctagaagg72361atacaagaga cccccgccga agcctggcac ccatcaaggg ctaggatgcc cacaatttgt72421ccagcatgga ggaagcagca gggctcagcc atgccttttt ctttttcttt tctttttttt72481ttttttgaga cggagtttcg ctcttgttcc aggctggagt acaatggggt gatctcggct72541cactgcaacc tctgctcccg ggttcaagca attctcctgc ctcagcctcc cgagtagctg72601ggactacaga cacccgccac caggcccagc tgatttttta gatttttagt agagacgggg72661tttcaccatg ttagccagga tggtttcgat ctcctgacct caggtgatag cccgcctctg72721cctcccacag tgctgggatt acaggcgtga gccaccgcgc ccggctggct gagccgtacc72781tttagtggtt tccacttctt ctagtaacac ctgggagata tagtggatgc tcctcaggta72841ttccgtgtat gcctcctgtg tccaggaaag agaaagagtg gggtcaggcc agtggaggga72901agtgactcca ttattccggc caggtgccct tgtgggataa gcaaggaaca ccaccacctg72961cttatctctg agatacaccc agagacattg agcctgcggt ggttagctgc agccggaaca73021cggttttctg cgcagggcac aggcagcatc gctgctctcc agacttggaa aagacatcag73081gacctgagag agagagagag ccagggacga gctggacctc attttctcct ctgactgcct73141ttctctccag cacaggaact gatattctca ttcagtgggc ctggggaagt gacctcagca73201gtttgctgtc atttttcatt ctatgcaatg tttccttctt gtgcaactca agggtgattc73261attcagtggc ttaaaaaaat acttttctga gaggataaag ggagtgatac tggaagtttt73321caaatcactt cccggtttaa gttccgagaa atccctttcc cagaacctgc gagttacgta73381tttccagtgc acacaggagc ctagaactgt tacctttcaa ggctgtgtga cccctctgac73441aaggcacagg tcattggaac cacagaatgg tggtttgttt tgttttgttg agacagagtc73501tcactctgtc gcccaggctg gagtgcagtg gcaggatctc gagtcactgc aacctccgcc73561tcctgggttc aagtgattct catccctcag cctcccaagg agctaggatt acaggtgtgc73621accgccacac ccagctaatt tttgtatttt tagtagagac ggggtttcac cacgttggcc73681aggctggtct caaactcctc acctcaggtg atccaccgcc tgggcctccc aaagtgtcag73741gattacaggc gtgaaccaca gcgcccagca atatgctatg tttttttgtt ttatccctag73801ctgttaagtt tcctatcttt aagtctatgc ctcagagaga aagacgaaat ataatccagg73861caggatttct gacttgcggg ggcacctgct gccctaaaaa ccaacttggg gaaagcagca73921gattcaggaa cgaggcttct cctaggcaca ggcaggctcc ttccctcctt gggcagccgg73981gtgtggctgg gccagcttct ctacctggtc gccaacctta gattcggccc catctcagga74041acgggccaca cccctccggc ctgcttttcc ttcctaagac gactgcaaac agcatcacaa74101atgagcaggt atgtggcctt ctctgcatgc cgtcccctgt ggcaaccgga ttaaatctac74161catctctgcc cagaaaaatg cacgaacgca cggacctgcY cgtcactcgg attttggaag74221gtggcctggc ggaccctcct gctacatctc ctctacaggt ggagcccagg ctgtgatgca74281ccggctcagc gagcggagtg gaaagccaag gtccgcagga tctgagctgt ccaaggtcac74341tgcacgcccg gctgcgcccc gcccccgtcc tcgcccggga accgcggccc gggatcccgg74401ctggcgcccg ctcccaagtc tccagcccga gccggccccg tcctcgccct ccccgggcgg74461gggtccctcc aggggccacc ctcagcggcc aagccccgcc cccgccccgc ctcacccggg74521gccggttgcc ggtgtccagc tcgatggccc cgttggcaag cttcatggcg ctctgcagcg74581gcttcaccgt gccgtccccg gccgcagcgg ccatggcgcc gagcggggga ggcggcggcg74641gtagccgagg ggctggacgg gcgaccggag gcccggaacg cagccggcgc tggtgctgga74701gccgcccgag cagcggaccg caggaaggga cgcggccgca cttccggcga cgggccccct74761ccgcgcgtac tgcgggcccc acgggtgtta gtggcggggg cggcagagtc cgggtgggtt74821gtcgcgacgg agccgggcct cttcgccgtc ttgagacggg gctggcgaga agggcccctc74881acggagttgc catgggcgtc taaccgcggc agccaggccc ctctctacgt gagaccccgg74941cccccctccc ctttctgcag cccgcccgcc acctgcgcgc cgcgtggcct ccgccggcgc75001ctgcccgccc cgcgcctccg tctcccacgg taagaggcgg tgagggccgc gtcgcctctc75061cctggaggac tcgtcgtgag gtgtcggcga gcccctccgc ccaccgcccc gccccaggcc75121agcgctccat tggcgacaga ggcgggcacg gccgccctcg ggcccgagag gggagcggta75181cgagcggggg cgctggcacc gcgggtggaa cctcggccgg cggggtcccg cccctggccc75241ctggcccgcc ccgcccgcct cgctttgctt ctcgctccgc ccctcccccg ccccgcctcg75301cttccagcgc gccgagcgga gcctaacgcc gggtcctcta ggaacctcgg gccgggcagc75361acccgcggga ttctgctggc gtcctccgct gccatgaagc gggaccggct gggccgcttc75421ctgtctcctg ggtcgtcccg acagtgcggg gcctcggacg gcggcggcgg cgtcagccgg75481actcggggcc gcccttccct tagcggtggg ccgagggtgg acggggcgac ggcgcggcgc75541gcctggggcc cggtggggtc ctgcggggac gcgggcgagg acggcgcgga cgaggcaggt75601gggtccgcgg cccgggcgtg gcgggttggg gtcgcggcag gggttgctcg tgggaggagc75661cgcactgacg cccggaagcc ggcgcggcct ctcctccacc cggccaaggg cgtagcggac75721tcggggctgc gtcgggatag gcgctgggcg gccggctcgg ttttcctcgc gctccgtgac75781gccggcgtcg ccggcccggc ctccccggcc gcgggctcgg ccagaaggga cccggcgtga75841ggagcgctgt caccgcgggc aggtcgccct ggggtgcccg cgcgtgggaa tcgccctccc75901ctgcttcgtg ccctcgcgga gccggaaagg aggcggcacc tgcaattggc agacaggaaa75961ccgcggcgtt tcctggaggg cgcagcgcct gccccgggtc tcgcggtcgc tgggaggagg76021gtgcagttcc aggtttcact cttgaagttg cttcgctgga gaggccttag aggcctttga76081aatttgaaag atttcttgga ccagcggcca catcccccac agggcggctt gtcccgccga76141gagccgcgcg cccgagccgc ttgctgtgtc cgggagccgc gcgaagggcg ctgcaggcgc76201tgccctcgga cctggagtcc gggacgccct gtgctcagga gcctcctttg ccagctgcta76261acacttcctg ccgctctgtg caggagcagg ccgggctctc gccatgggtc actgtcgcct76321ctgccacggg aagttttcct cgagaagcct gcgcagcatc tccgagaggg cgcctggagc76381gagcatggag aggccatccg cagaggagcg cgtgctcgta cgggacttcc agcgcctgct76441tggtgtggct gtccgccagg accccacctt gtctccgttt gtctgcaaga gctgccacgc76501ccagttctac cagtgccaca gccttctcaa gtccttcctg cagagggtca acgcctcccc76561ggctggtcgc cggaagcctt gtgcaaagta cgccctagtc tgttcagagc acgttcaggc76621tgtcagtact gcagtgtgac gggtgttgag agagggacag ggcgtgcctc cgcgggagcc76681tctgggtggg gggaatgggc catgcccggg ttcagtgcca acagccctgg gactgttgtg76741gagagatgaa ctgggaggag ctggggtgag ggaagagaaa gttgcctatg gggaccgctg76801aggtttgaga tctcgagagg gtcccgtacg acgagcgctg tgaacctccg cctgcttgtc76861ctgctcatgg ccacactgat cctttgcagg gtcggtgccc agcccccaac aggggcagag76921gagggagcgt gtctgggtga gtcctccccc ggtggagggt gggctgggtg ccgaccagcc76981gtggatctga catctctgtt gactctctgc agtggatctg atcacatcca gcccccagtg77041cctgcacggc ttggtggggt gggtgcatgg acatgcggcc agctgcgggg ccctgcccca77101ccttcagagg acactgtcct ccgagtactg cggcgtcatc caggtcgtgt ggggctgcga77161ccagggccac gactacacca tggataccag ctccagctgc aaggccttct tgctggacag77221tgcgctggca gtcaagtggc catgggacaa agagacggcg ccacggctgc cccagcaccg77281agggtggaac cctggggatg cccctcagac ctcccagggt agagggacag ggaccccagt77341tggggctgag accaagaccc tgcccagcac ggatgtggcc cagcctcctt cggacagcga77401cgcggtgggg cccaggtcgg gcttcccacc tcagccaagc ctgccccttt gcagggcccc77461aggtaggagg cacctcttgc tggtgctaga ccaggatgtg tgctcctcag tSgggcaggg77521ttttcagcag aaagtgaatg tctccactgc tctaggtggt ggctggggtg tggtgtgaga77581aggagcagag cttggggctt ctgcctgcgg ctgctcacca catccagaga gcagggaggg77641gcggccagcc tgtctccgcc tcttgctcct ctgtgagcag ggctctgttc cagcagtttc77701tgtggagaca cgctcatcag ccgtcgagtc tttttctaag cctgtgcttg gggctgaggg77761gactgaggaa gagggaagaa tagggccttg tctgtggatg tgggagaaaa gaatggccgt77821tgcctctgcg gctgctgccc tgggccagct gcaggtcatt tcctggcact cagccagccg77881cctggtgaca ccctggtgtg ggccagtgtg gcgcagcttt ctctgctgac aggctgaggg77941gaaacacaag gcccttgttt gggccccaga acccatagca ctgtgggctg gtgcagctgc78001tcctggatgt tgttttagga gggccagggc cagtttccct ccggcctctt gtcctgtgtg78061tgctctgtgc acctgtcccc acctgagcac tcagttccca ctttgagtct tgaggggagc78121ttccgaaaag cgtagctacc tgtaatccca gcactttggg aggctgaggt gggtggatca78181cttgaggtca cgagatcaag accaacctgg ccaacatagc gaaaccccat cgctaataaa78241aatacaaaaa ctagccaggt atggtggtgg gcatctgtaa tcccagctac tctggaggct78301gaggcacgag aatcacttca gcctgggagg cggaagttgc agtgagccga catcgcacaa78361ccgtactcca gcctgggtga cagagcaaga ctctgtccca aacagaaaca cagctaccca78421gatagtttag ctagatacgg gtgcacagat gagacagtgc ggctcggggg cctcaggtag78481gaacgggatt gtgagtgtca catcttagag ctgcttggct ctgtaaactt gtatgcagtg78541tgttgagaac aattctatct atctgtgtat ctatctacct acctacctat ctatcttcct78601acttctttgt ctctctctat ctacctacct atctgtatat ctatgtatct acttatctac78661ctatttatct atttatttac ctacttatct atctatctat gtgtttattt atttttgaga78721cagagtctcg ttttgtagcc caggctggag tgcggtggtg ccatcttggc tcaccgctac78781ctcagcctcc cgagtagctg ggatgacagg cacatgccac cacgcccacc taatttttgt78841atttttagta gagatggggt ttcaccatgt gggccaggct tggtctcgaa ctcctgacct78901cgtgatccac ctgcctcagc ctcccgagta gctgggatga cgggcacgtg ccaccacgcc78961cacctaattt ttgtattttt agtagagatg gggtttcacc atgttggcca ggcttggtct79021cgaactcctg acctcgtgat ccacctgcct cagcctcccg agtagctggg atgatgggca79081cgcgccacta cgcccaccta atttttgtat ttttagtaga gatggggttt caccatgttg79141gccaggcttg gtcttgaact cctgacctcg tgatccacct gcatcagcct cccagaatgc79201tgggattaca ggtgtgaacc accgcgccca gccaagaaca atacttttta aaacttattt79261taggccaagt gcagtggttc acgcctgtaa tcccagcact ctgggatgct gaggtaggcg79321gatcacgggg tcaggagatc gagaccatcc tgactaacac ggtgaaaccc catctctact79381aaaaatacaa aaaattagct ggatgtggct gtggttgcct gtaggcccag ctacttggga79441ggctgaggca ggagaatcgc ttgaccctgg gaggcggaga ttgcagtgag ctgagattct79501accactacgc tctggcctga ccgacagagc aagattctgt ctaaaaaaga aaaaaacaaa79561acaaagaaaa cttattttaa gtagagacag ggtctcactc ttttgcccag gctggtcttg79621aatttctgtc ttcaagtgat ccttctgcct tggcctccca aaaggctgag attataagca79681tgaaccttca ggcctggccg agagaacagt aatatttatt tatttattta tggagtctcg79741ctctgtcgcc accaggcctg gccgagagaa cagtattatt tatttattga tggagtctca79801ctctgtcgcc caggctcgag tgcagtggca caatctcggc tcactgcagc ctctgcctcc79861tgggttcaag tgattctcct gtctcagcct cccgagtagc tgggattaca ggcttgtgcc79921accacaccct gctaattttt ttgtatttgt atttgtattt gttttgagaa ggagtttcac79981tctttgttac ccaggctgga gtgcagtggc acgatatcag ctcaccacaa cctctgcctc80041ccaggttcaa gcgattctcc tgcctcagcc tcccgagtag ctgggattac aggcatgcac80101cactatgcct ggctattttt gtatttttta gtagaaatgg ggtttctcca tgttggtaag80161gctggtctcg acctcccgac ttcaggtgat ccgcccaccc cggcctccaa agtgctggaa80221ttgcaggcat gagttaccac gcccagctca attttttgta ttttttgtag agacggggtt80281taaccatgtt agccaggttg gtcttgatct cctgaccttg tgatccaccc gcgtcagcct80341cccaaagtgc tgggattaca ggcgtgagcc actgcacccg gactgagaat agtaattttt80401aattatttta tttttttgag tcggagtctt gctcttttgc ccaggctgga gtgcagtgcg80461cgatctcggc tcactgcaat gtctgcctcc cgggttcaag cgattcttct gcctcagcct80521cccaggtagc tgggattaca ggtgtgtgcc accatgcccg gctaattttt gtatttttaa80581tagagatggg gtttcaccac gttggccagg ctggtctcga tctcctgacc tcgtgatcca80641cctgcatcgg cctcccaaag tgctgggatt acagacatga gccaccgtgc ctggcctagt80701aattttttaa atgtctcaga acatccacca agtccaggct gtcctagaag gactggatcg80761gagtcagtgg aaggactggg ctttctggtt ctgctgttcc tgagggttct gatgcttgtt80821gctgcctgtg tgcttctgga cgttgggctg gcagggcagg atcagaggct gcatagcacc80881cttggtgggg atggggccaY ggtggggaga acttgagggt cagggacccc agtctccctt80941ctagtcatca atagtagaat catgcctcct tgcaggtgag acctttctgt gctccgtgtt81001ggtaacagga gccttgttct ttgtttctca gggcagttgg gtgagaagca gcttccatct81061tcaacctcgg atgatcgggt aaaagacgag ttcagtgacc tttctgaggg gtgagagaag81121agtgggttta aacagcctaa aatttctcct tcaaaaacca tcagtgacct ttctgagggg81181tgagagaaga gtgggtttaa acagcctaaa atttctcctt caaaaaccaa acagccatcg81241tagggtcggc tgcccatgcg ctggtaccaa actaagccat cacaggggcc ggctgtccgt81301gccctggttg ggacacgcct cctctcgagg tggcgggaga ggaggggctg gggagcttcc81361gaggcttctt gtctctgatt gatggtttcc tgggtgccag agagcactcc cagacctgag81421gcagctccag agggagcctg atagggtttc tgacccagag cagcctagaa atgagtttgt81481tgaaaccact ctggctgggt cctttgcttc tgctactgtt ttagggccat agttttgtgt81541tcgctccaga tgtttctgga gtagagaaac cccgacttta gcccttgact atgacactgt81601tgtggcttga agccgaggct ctgcgaggag gtgttgcagg aggacgggcc tcagaaggga81661cctcatcagt gtctgcaaac tccctgggcc acccctgcct ctgagccaag cgccattcag81721ggcagctgtg ccaggctggg gtttgcaccg tttccaccct ctcctgtaaa ccttggctgc81781aacacagatg cacctcatcc aaatgaggac accccagttt gggggccaat cccagtggga81841tgtgcagtga tgaaatcggg caggtgccag gaggcacctg gggtggctaa gactgtcctc81901aggaccacca ggacacaaaa ccgcagctga ggttatagca taaaggccac aaatcttttt81961tttttttttt ttttcctttt gtgaactttt ttttttagac cgagccttac tctgttgccc82021aggctggagt gcagtggtgt gacctcagct cactgcaaac tccacctccc gcgttcaagt82081gattctcccg cctcagcctc ccgagtagct gggattacag gcgcccgcca ccacacccag82141ctaagttttg tatttattta tttttttttt atttgagacg gagtctcgct gtgttgccca82201ggctggagtg cagtggcaca atctcggctc actgcaagct ctgcctcccg ggttcacgcc82261attctccggc ctcagcctct gagtcgctgg gactacaggc acccgccacc acgcccggct82321aattttttgt atttttagca gagacggggt ttcactttgt tagccgggat ggtcttgatc82381tcctgacctc gtgatccgcc cgcctctgcc tcccaaagtg ctgggattac aggtgttagc82441caccgcaccc ggccaatttt tgtattttta gtagagacgg ggtttcacgg tattggccag82501gctggtcttg aactcctgac ctcatgcgat ccacctacgt caccctccca aagtgctggg82561attacaggag ttagcctctg cacccagcca cttttgtgaa cttagaagtg actcatggca82621aacccatcag ttgcataggg ttcgcagtta tggggtgagg tggtcgattt ctcaaagtct82681gcttagcagg aagctacctt tgagcgcctc caccccttct gggccgctgt ggaaagggct82741gtttccaagt ccagtgtcct ggtgcccaga gcacatggct tgtggggatt gctggcttgt82801gagcctggct ggccctttgt gttcggaaac gggattggaa atttggggat tttagattta82861agacattttc agaagaactc aaatgtggga catgggggtc catttggtat ctttaggcag82921gaggtcgtgt tgggtctgtc actgcccagg cccagggctt tgctgaagat tctctcttaa82981ttcctagaga cgtcttgagt gaagatgaaa atgacaagaa gcaaaatgcc cagtcttcgg83041acgagtcctt tgagccttac ccagaaagga agtaagtggg cagcccgggg tctgctggag83101gcatccgttg ggcgacctag acacccgcct gtgctccagg gcctctcccc ggtgcccact83161cagttcactc tctcgtgtgc ggatctcccg agcccagtga aacgtggctt ccctcagtga83221gcgggaggcg ggggaggggc aggcgggcag gtgagaaggt gggtcctcct ggtgctttgg83281agctgggacc tggtctcggc actgtgaggc tgcctcagag tcccagcagt ggatggcagg83341tgtctctgtc gacagaaccc cgccttgctt gagaaggagg ctcttacgtc ccttctgccg83401gtcctgccct gggttcacac cccagtgtac ccccgaagcc cccaacccgt cgcttctgct83461ttgtgtgcgt ccccagaggc ccgaggaccc agctctgtca tcctcccagt tctgacccca83521tcactgatgt ccatggccag gcagccacgg tccacttccc tgttgaatga cttctgtgag83581cttgttttga ctttgttttt agggattttt attttctctc ttcagttaga acacaggccc83641tggtagtgag ggctggtttt cacttgtgct gttttctttg cagccctata cccagcagta83701cccagaacac tcccaggtag ttggtgactg acccagtccc tgtccttgag catttcacag83761cccagaacaa aggtccagag ggcaggtggc tctgagcacc accgtcagcc actagaaccc83821tgctgtgcag actgagaggt caacggcctt ttttcatgga agttctgatg ctgttaagtt83881ctgaaaagag aaacgggagc agtggaggag gccYggagag agatgcgaag cagagggcag83941gactgagcca gaggggctgc agcagcttca gcatgaacag aaaactggtc aggcccagaa84001aaggtgcccg ggggtcccta cccccaggct tgttcccagg agtcatggct gggagtgccg84061cctgaggctg ctgtacggaa gcccccactc cgtgaccctc Ygggttgagt ctgaatccca84121ggacacctga gaatgtgtct ctggcttgct gcccagaagg tctggggctg ctggtgtgag84181gagggctggg agctgggatg ggaggagcca ggcgagcctt ttccggatcg aggcagattc84241aggaggggag ggtcctctct gctgagcccc tgctgctgtg acagggacag cacactcaga84301ctgtgtcact agtccctctg ggaccccagg agtgcagaca aagacgtgct ttggaacaga84361gacacgggag atacgttttc agaggtctct aataaaaggg gtgacttcag tggaaaccgc84421tgcgcaggag gcaggactgg gtggtagcaa gcatggcctc ctgactgcgg cgagtaccag84481gggcacttgg ttccccagct gccaccgtga gacccccctg ggaacccaga agggcctctg84541cacagccccc ggcccgtgtg ctgcctccca gtcttgtcaa ccctgacctt ggagttctag84601cgggcgtgcc cgcctgtgtc ctcagctggg ggcgcatgag aagagacttc tgtgatgtgt84661taatgagggg tcagtggttc ccgccagagg ggaaggccag ggcctgcacc cccatgggga84721aagaactgac aggcagcagt gaggcctgag cccagcccac ccggtccgca aagattcccg84781tcccaaacac gagggcacag agactgcggg tgctctgaag cctcctgtgc tgaggaggtg84841agtggtcatt gtgggaaYgS catggccagc cactcacaaa caaggggcca cccaatccag84901gcaggcaggg caggtgaggg ggacatgtgg ctaggtcatg aaaaccagtt attagggaca84961gggagagaag gYgcagcctg tgcgaagtca acagttcttc tcagtgcgtt tttgtcttgt85021ttctgagatg gagtcttgct ctgttgccca gactggagtg cagtggcacg atcttggctc85081actgcaacct ccgcctcccg agttcaagtg attctcctgc ctcagcctcc tgagtagctg85141ggattacagg catgtgccac catacctggc taatttttgt atttttagta gagatgggat85201ttctcccacg ttggccacac tggtctcgaa ctcctgacct catgatccgc ccgcctcagc85261ctgccaaagt gctgggatta caggcgtgag ScaccgcgSc ggcctcaatg cactctttaa85321agagaagggc agttagcgac agggtctcat gttgttgcct aggctggggt gccctggcac85381gatcatggct gactgcagcc ctgtgcgctc aagtgagcct ctcacctcag cctcccaggg85441tgStgggact gccaacatga gccactggac tcggcagtgt aaactacttt tttcatgcta85501ttgcatatgt attgttcatt gataatatat agaaacataa ctgggttttg tgtatggatt85561ttgtgccctg caactttgct gaaataatta attagggttt tttgtatgtg gatctttagg85621gttttctacg tagaaggtcg tgtcatttgt gaatagggag aatttgtctt tgttttcttt85681ttcttgccca gttactctgg ctagggcctc cagtgctgcc ctgagagcag tggtgagaga85741atccgtatMt tgttcctgac cttggaggaa gagctttcag tccttcacag ttgagcgtgg85801tgtctgctgt gggtttcatg tagaggtttt atcgtggtga agaagttcct ttctgttctt85861Sagatatttc cttaactatt cttacctgga tccagaaagg aaccacaagt gcgtctctct85921ctttaagtct ccgatagacg cacactggaa gctgctgcat ttgctggtcc ctcagggact85981tgccagacca actaaaatgc acgaccccag acttttggag gacaaggtct cctgcccacc86041atggtcccag ccagcagctc caggaacaca ggctgggctc cctgcagtgg aggcggggtt86101gaggaatgct ggggtggcag ctactcaccg tgtgcttccc agggagcatc acctgttcct86161ctgaagccct gcctgggtgg cttgagcttg gaatcaagct gacccatttt gagagagtgg86221attctgagct ttttcctagc gtaaggtcgt tttgataaag gagtggtcct ttggacatcc86281ctgttctgcc gtcctgatac ctctagttgt gggttttatg tgtagcacag ggtgaagctt86341tagttctcag aacccacccc tgctgtaccc tgtgccctcg ccctccgctg tgtttgccct86401gaccctcctg taccctgcgt cctcgccctc tgctgtgctc acccgaccct gctgtaccct86461gcgccctcgt cctctgccgt ttgcccctga ccctgctgta ccatgccctc gccctctgct86521gtgttcaccc tgaccctgct gtaccctgcg ccctcaccct ctgctgtgtt cacctgaccc86581tgctgtgtcc tgcgcccttg ccctctgctg tgctcgccct gaccctgctg taccccgctg86641taccctgcgc cctcgccctc tgctgtgttc accccgaccc tgctgtaccc tgcgccctca86701tcctctgctg tgccctcccc ctctgctgtg ttcgccctga ccctgctgta ccctgcgccc86761tcgccctctg ctgtgtttgc cctgaccctg ctgtaccctg cgccctcgcc ctctgctgtg86821ttcacccctg accctgctgt gtcctgcgcc cttgccctct gctgtgctcg cccctgaggc86881ctcctgtcca gagttgctct ggctttggtg gcttcaggta gattctgagg gtcagtcagc86941acagaagcaa ctcagggaag cttcagaaaa gaccatttct caggttggag agacaaaaaa87001cattttgcaa atggagatca gaaaccattg aatttgggaa cctctttttt tcagagtctc87061tggtaagaag agtgaaagca aagaagccaa gaagtctgaa gaaccaagaa ttcggaagaa87121gccgggaccc aagcccggat ggaagaagaa gcttcgttgt gagaggtgat gcctgcaacg87181cgaggctcgc cctgcctgtc ggggccgggg ctccatgcat gctcttcatt gcagggagga87241gcttcccacc atctacaagt gtccttacca gggctgcacg gccgtgtacc gaggcgctga87301cggcatgaag gtgagcactg gctgtgcctg acccaggccc ggcagctgtc agtgtgaacc87361tgggggaggt agcaggtgtt tctgcagcct aactcagact tgYgcccaag gacactttga87421cctaggttaa ccgagacctg aagcagtcct agccagtgcc atccttgggg gtagatgtgR87481ggccagactt tcacttgaga caccccccgg ggggtctaga agtttcttgg agtgaagctg87541tgaaggagcc agtggtgtcg ctggaggagg agttggatct gccttccagg ggcctcagct87601gtcacctact gagggctcgt gccatgtggc ccgggtgcgg ctctgagggt ctctcaccga87661gtctctcctt cagaagcaca tcaaggagca ccacgaggag gtccgggagc ggccctgccc87721ccaccctggc tgcaacaagg ttttcatgat cgaccgctac ctgcagcgcc acgtgaagct87781catccacaca ggtacgccta tcgccagtgt cgcccacgcg ggtgacagcc aggggcacgt87841gacctgcttt ccagtcttcc tcatacccat ccccgcccca agagttgggg gtgcgtgaag87901gtggctcatg gggtctttgg tactcagtca cgttggtaga tgaaggctag agtagggcgt87961cagtgtgaat ctgcccttcc ctgtgatgga aggaattcct gacttcccca ggatctaggc88021agtttgtcaa aactaatagc caagcaatct tggtgtcctt ttgcaaattg gtcattttat88081tttttaattt ttttttgaga cggagtcctg ctctgttgcc caggctggag tgcagtggca88141ccatcttggc tcaccacaac ctctgcctcc caggttcaat caattctccc gcctcagcct88201cccaagtagc tgggattaca ggcacccgca accatgcctg gctaattttt gtaatttttg88261gtagagacgg ggtttcacca tgttggccag ggtgatctct aactcctgac ctcaagtgat88321ctcagccttc caaagtgcag agattacagg tgtgagcaSt gcacctggcc caaacaagtg88381tatcttaaaa ggcaaggcag aaggcatgaa ggcacaccag ctctggggag ctgcccacgt88441ggacaggcgg gaggcacgtg tggttctgcc tYttcacRtg tgtgtaggtc gtgtcattaa88501gaccggaggc ggatggggga attatgactg agggtgctgt ctctgtgccc gtcttatttt88561tcacaatttc tataatgcac aagttacttt tataatccaa caaggatatg ttttcaaaat88621gacaaacttg gccgggcgtg gtggctcaca cctgtaatcc cagcactttg ggaggctgag88681gtgggtggat cacctgaggt cagcagttca agaccagcct ggccaacatg gtaaaacctg88741gtctctacta aaaatacaaa aattatctgg gtgtggtggc acacgcctgt aggcacagct88801acttgggagg cggaggtgga ggttgcagtg agccgagatc acaccactgc actccagcct88861gggcaacagt gagacactgt ctcaaaaaaa aaacttgttt gaaccacgtg gacccacata88921tccatggtca gggactcgaa tccctggtat tcagagggct gccttttcct aagtgcaggt88981tctgtagggt ctttgcaggg ctaacttcgg gacttcagta tgagcagatt ttggtaaggg89041gtgggggggg gcctggaacc agctgaggac aactgtatat tgatccttac acacacagtt89101ttccttggtt ttgaacttat tacagaataa gaattattta ttcttacttg aatttactga89161agtatttttt cattaggatt gcttttcact gctgggtcat ctcccgacat gcgggttgtt89221attttttggg ttctccgaca tctctcctga tggcagcact tccttgggtt tggtccttca89281gactctgcat ctgtccggtc actgatggtc acaaccagca gttccttaaa aacctgaaat89341tgggccgggt gcagtggctc atgcctgtaa tcgcagcact ttggaggcta aggcagtagg89401attgcttgag gccaggattt tgacaccagc ctggRcaaca tagtgaggcc gtctttacca89461aaaacttttt ttttaattta ttgatggaaa cagtctcact cttatcacca tggctggagt89521gcagtggcgc gatcttggct cactgtaacc tccacctccc tagttcaagc aattctcctg89581tctcagcctc ctgagtagct gggattacag gcatctgcca ccacgccggg ggtgtttttt89641tttgtttgtt tgtttgtttt tttgactgag tcttgctctt gtcgcccagg ctggggtgca89701gtggcacaat ctcagctcac tgcaagctcc tcttcctggg gtaaagccat tctcctgcct89761cagcctccca aatagctggg actacaggtg ctggccacca cacccaacta atttttcgta89821ttttcagtac agacggggtt tcaccacgtt agtgaggctg gtctcaatct cctgaccttg89881tgatccacct gcctcggcct cccgaagtgc tgggattata agcgtgagcc atgacgcctg89941ggtaattttt ctatttttag tagagaggca ggttttgcta tgttggccag gctggtctcg90001agttcctgac ctcaggtgat ttgcctgcct tggcctccca aagtgctggg attacaggca90061tgagccaccg tgcccagccc ccaatttttt tttttttttc cgggacagtc tgtgtcgccc90121agcctggagt gcagctgtgc aatctcagct cactgcaacc tccacctcct gggttcatgc90181aattctgctg cctcagcctc ccgagtagct gggactacag gtgtgtgcca ccacacctgg90241ctaatttttg tatttttagt agagacaggg ttttaccata ttgattgggc tggggctggt90301cttgaactga ctKcatgatc cgcctgcctt ggccttccaa agtgctggga ttgagcactt90361tgtgtaagcc actgcaccgg ccccccaaat tttttaaatt acctgggtgt ggtggtgcac90421acctgtggtc ccagctactt gagaggttga ggcaggagga tcacttaagc ctgggagttc90481caggctatag tgagccatga ttgtaccatt gcacttcaac ctgggcaaca gagcaagacc90541ctgtctccaa aaaaaaaaaa aaaaaaagaa tcccttgaac ctgggaggtg gagtttccag90601tgagctgaga ttgggctgtt gcactccagc Stgggcaaca gagcaagact caagtctcaa90661aaaaaaagaa cagcgtagag taaaatgcca agatagggtt tcccttcgaa gagaaaagtc90721tgtcttgtga agcgtttcag ggcttgacag atgtggaggc agcacagggt gagtgggtct90781gcatgtccac atgtggcatt tgtggactag caagtgagat ctctttgttc aagggtgtga90841gaaagatttc cagagccagt acagcaatgc atacceccag accctgggta tggacccctc90901tcatatggag atgaattcta gagaacagcc atagctgtga caatcagtat tctatacaac90961cttagtgata cggcttcctt taagaatttg taggccaggc acggtggctc acacatgtaa91021tcccagcact ttgggagacc gaggcaggca gatcacaagg tcaggaattc aagaccagcc91081tggccaatat ggtgtaaacc ctgtctctac taaaaataca aaaatcagct gggcgtggtg91141gggggcgcct gtaatcccag ctacttggca ggctgaggca caagaatcgc ttgagcctgg91201gaggcgcagg ttgcagtgag ctgagatcac gccaccgcac tgcagcctgg gtaacagagc91261gaaactccat ctcaaaaaaa aaaaaagaca agaatctgtg gttaaggaaa tagctttctg91321aggtttcttt aaaaaccatc ctgaaatgca cacagctgat gaagccacgt gacagtgtat91381aaagcagttt aaagatctta ataaacgagg ccctcatagg ccccttgctt gggcccactg91441catggtgaac catgtgcaga aatgtcttcc cagctgtgat ggtttcacat tgtcatcgtc91501gtccccccgg gaggttggag catcaggggc ctggactcac tggactctcc cctctcagag91561gtgcggaact atatctgtga cgaatgtgga caaaccttca agcagcggaa gcaccttctc91621gtccaccaaa tgcgacattc gggagccaag cctttgcagt aagtgtgagt caggaccccc91681tcccagggct gtggccctcg caccttctta tctgcctctg tcccccaggt gtgaggtctg91741tgggttccag tgcaggcagc gggcatccct caagtaccac atgaccaaac acaaggctga91801gactgagctg gactttgcct gtgaccagtg tggccggcgg tttgagaagg cccacaacct91861caatgtacac atgtccatgg tgcacccgct gacacagacc caggacaagg ccctgcccct91921ggaggcggaa ccaccacctg ggccaccgag cccctctgtg accacagagg gccaggcggt91981gaagcccgaa cccacctgag gacggcagtg aggatgagca cctctagcag cctggactcc92041gcagtggctg tgtcagcctc acccttcgtg tgcacccgca tgggagggtc ggagggtgct92101gcccgccctt ggtgctggag gcgggcttgg tgtccggctc aagtagcctt cctctgctct92161gggaccagtg gtttattttc ccgcaaacgc tgagtgactY ggggccggac agttcataaa92221taattgattc ctttccccac taaagcagtc gaggagattt gtaatccact ttttagtgca92281acaagagctc catgttatgc ttgtaataaa ttatttacac gggagctggg ctggtgtgca92341gtggcaggtc ccgtcagaag agatgaggct cctgggacag gtcagcgtca ggggcagcct92401gctgtctgct ctggagggcg gcgctcacct ctgggtcgca gtccccacga tcagccagca92461gctgtgagag aggagcaggt cctcagccca tgccgcccac taggcctcag accacagggg92521aggggctctg gcagaaatag tcgagttgta ttgccagcca ggcaggcaca tggcccaggc92581agctgtcaat tctcatgtcc cccacRtggc ccaaggtggg catcttgacg ttacctctgc92641cacgtgtgag aagctctttt tcgggcaccg aggtattaac tgcagcagaa aaagacgagc92701ttttgttatc agttccacgg ggttgcccta gagagaaaac aggcaaactc acaggttaga92761agacatacag aaacagggct ggtgtgtccc ccatagtctg catgctgtgc cggaacattc92821tttggcagaa ggagcctccg gctgggggga gctcccctgg aggtgggact ggcccttgca92881cctgcctgac ccttgagctc caggctcctg ccagctggag gtgaaactgt gcttgtatcc92941ccagccacga agagctggac cagcttcaag tacatgtcca cagcaacatg caggaaggcc93001tcttccctga tggccgcgtc ttcatggaag taggagagaa gactagaggt aaagacatag93061tgacaaatgg ctacagactg ctggaaaggt agcaggtgat gccaagggat actgctcatc93121tgtggagcag aggcacagac aacccttccc atctggcggg acccagaggt gctgagatgg93181gggtctggga aacactgccc agccctgacc agccctgtgg gtggaggtac ctgtaaaaag93241cgaaaggcag cagcctggtg tgctgatccg gggccacacg gaggaggagc cgccccagcc93301tgaggtctgc aacaccaaga aRtggctcag gcaactctgg acatctctgc ctattatcag93361tgctggggac acccctgggg gtcgggacgt gtaccctggg aggcctggct gtggggatag93421tgtggggcga acagcctgag ctgaggatac ccaggtacct gtcagcagct gggagaggat93481gggggggtcg acctcttgca ggagggtggg tgtggtgcag agagaggcag tccccatgat93541aggcccattg gtcctggggt tgaccagtga gccagtaaat tatcttattg ctttaaacaa93601gtttgtgctt aatctgtccc aactaaaatg gagcttataa acttacttag caaggaacct93661caaggagggc tcgttcttaa ccatttgcaa gatgcctctg aaaagagcgg ccctccgcat93721ttgtgcctca gcagcgtgtt tcttaccact ctctgtcaac tgaaagagtg ccagccagga93781tatcttcctc ttctctaaac actcgaggat tgctgcacaa acgtggaaag cctttggcag93841gtctgtggtg ctctgtaaac cgcaggagac caaccctgag aatggccgac ctggtgctcc93901catgggtagg agggtacagc cctcagcaca gaagagggca tttcctcttt gcttattgta93961agtcttaaaa ctggtgacag ttttacctat agaaggtaat actgggcctc tggacagaag94021aggctcaggt aggaggccag ggacttcgag cacccacacc aaggctgctg caccacgtcc94081tcaaataaga cattaaaaga aaggcccaca ggccggatgc agtggctcat gcctgtaatc94141ccaacacttt gggaggccga ggtcggcgga tcactgaggc cagttcaaga ccagcctggc94201aatatggtga aaccccgtct ctactaaaaa tacaaaaatt agccgggtgt gacagactca94261cgcctgtaat cccagctact cgggaggctg atgcaggaga attgcttgaa cccaggaggc94321tgaggttgca gtgagctgag atcacaccac tgcactccag cctgggtgac agagtgagac94381ccccatctca aaaaaaaaaa aaaaaaaccc acggcctggg agttctcact cacacttccg94441caaacacaag gagctcctga gctagtctgg aaaccctgac ttggaagctg gctgcctggt94501gcccctgcct ggcccacagt gggagaggac accttggctg gtaaggtctg acttacattt94561gaggtcagat gtgacgacag caggcccatc aaggagaaga agaaaaggaa aaccaatagc94621tgtaaataaa aacgtgcact tattattaca ttaaaattac ctgtgctgtc attctaaata94681aggctgacac attcctcttt aattgaaatt ttttacatct aggccataaa tcctttaagt94741ggatcttaga aaactttcca atcacttcta gagagacagc ttaattgaga attaattact94801actggctggg tcatttcaca cttgccttta aacaaagcta gaaacagttt gaaaaagtga94861caagagccag aaagagaaga ctgctgcggt ctccttggct gtgcgcagtc ccaactcagc94921atcatctccc tgcaaggcct cctgtcagaa gtgtttgtaa aatccttgaa gataagccca94981caggctccag cctaaagtag ggcctgactg ggcagggtct ctgtggacac tcctcgctgc95041acactagggt cctaaggccc tggagccact aaaatccctt ctaacaaaac tacacaagaa95101acctgaagac gtactcagaa gaacccctgc agggtggtta taatcaccaa aaccctgggc95161gctgctgtcg atccagcttt gagctggtgt aagttgtgtg ctgacatggc acctatgcgt95221tgcggatctc gggctgggca cagagggctg ctgcctccag ctctcttgtg ggatgcttgt95281ttctgttcct gttctgcaca caccaccctt ggctccctgc acagagagct gtgtgggtgt95341cactggtctt gtgggggcag tcaggctttc cagggtgcag caaccaggtg ggcagtgggt95401ctgagtcgaa catggtccct ggcctggaat tcaagagcag cctctcccac tcctttccct95461atgcctatgg ctgcagacgg tcccatcctt gctgcctctt gtccccagca ccccaggtct95521gagctgtctc actgcaagct tatctggggc tgtccctcag cacatgaatc aaggctactg95581cagcatgaac acagaaggca ccacttgaag gctgacagca tttacgctgg gctgtgatga95641ggataaaaat aagctgaaga aacgcacttc aagcctgggc aacacagtga gacccccatc95701tcttttaatt tttactttaa acaaattttt tttttttgag acaagtctcg ctctgtcacc95761cagactgcag ggcagtggcg tgatcttggc tcactgcagc ctctgccccc tgggttcaag95821tgatcctcct gcctcagcct cctgagtagc tttacaggct atgggattgt aggtgcgcac95881caccacaccc gactaatttt tgtgttttta gtagagacag ggtttcacca tgttgcccaa95941gctggtctca aactcctgac accaagtgat ccgcctgcct cggcctccct aagtgttggg96001attacaggca tgagccacca tgcctggact acagaccccc atctcagaaa aaattacaaa96061attagccggg catggtggca cacacctgta gtctgagcta ctccagaggc tgaggtaaga96121ggatggcttg agcccagaag aggctgcagt gagctgactg caccactgca ctccagcctg96181ggcgacagtg agaccctgtc tcaactacat acaaccattt aaaaaactga aattattatt96241tttactagta aacttgatag tcaaagtctt actccaagcc agctgggcgt ggtggctcac96301gcctgtaatc ccagcacttt gggaggccaa ggcgggagga tcacggggtc aggagatcga96361gaccatcctg gctaacacag tgaaaccccg tctctactaa aaatacaaaa aaaaaaaaaa96421aaaaaaaaag tcttgctcca agccacatat ttgtctttag aaaaacagtt catcagacaa96481gcccagagaa atagcactga ttgaaaccaa gcttgcgaga aaataaatca gtaaaagaat96541ttcctatctt gcctcctctc tctcgcagtc cagcttcttt agctgcttcc tgatgttttc96601ttccctgact tgttgaatcg caaagtgcag tgcagcagct gagagccagt ccgggttggg96661tgcYggggag gcagcctcag gggagaggaa actgggacag agagaacggg gtcattgcag96721ggccttacaa ccatacaacc acgccataga aaccaagtcc ttattcccac ctgtcacctt96781tggggcctgc ctttccatca gaggacagag aagggtttca ggaccatcag aaactaggtc96841tgcatccccc aggtccacgt gagagtgtgg gcaggtgctg gtgctgcccc cacRgcatcc96901tggcgttctg agagctgctg actgagcagg cagctggcct ggtcctgtct accttgccct96961gtgctcttgg gccccagctg cctacatttt ggaagaagag gcctcatttt cctcagttgc97021ccatgcctgg ccctgtggat caggcattcc ctcctacaac agagaacaaa ggactgcctg97081gctctggccc cagtgtggtg tctgagtttt ctggctggtt aaaaaattgg taggaaatta97141atgaaacaat gctgtggttt aagagacttt aaataagctg ggcatgacgg ttcacatctg97201caatcatagc acttcaggag gatcgcttaa gcccagcagt ttgagatctg cctgggcaac97261acagtgagac tcaatctcaa ataaacaaaa acgaggcagg gcgtggtggt tcacgcctgt97321aatcccagca ctttgggagg ctgaggcagg cagatcacct gaggtcggga gttcgagacc97381agcctgacca acgtggagaa accctgtctc tactaaaaat acaaaattag ccaggcatgg97441tggctcacgc ctgtaatccc agctactcag gagactgagg caggaggatc gcttgaacct97501gggaggcgga ggtcatggtg agccgagatc gtgccatctc actctagggt gggtgacaag97561agtgaaactc tgtctcaaac aaaacaaaac aaaaaacaaa acttcagaca atctgatctg97621tgtttgtaga attagaactg gaggtgtgac ctgctttctc cttttttgag atggagtttc97681tctctcattg cccatgctgg actacaatgg catgatctcg gctcactgca acctccacct97741cctgggttca agcaattctc ctgcctcagc ctcctgagta gctgggacta caggcgcaca97801ctatcacacc cagctcattt ctgtattttc agtagagatt gtttcacatg ttggccaggc97861tggtctcaaa ctcctgacct caggtgatcc acctgccttg gcctcccaaa gtgctgggat97921tacaggtgtg cgccaccgcg ccggggccct cttctcttct ttcagcagac ggcgagcagc97981caaaccctga ggggacccgg tgtggcatca gtgattcaaa actagtcaca aagtaaggtg98041ttctccaagg aggcaggatg ggtactagca gtgccctctg acgacccctt aagaacacgg98101cacccacatt cagcaggtgg agtggctgtc agctctgtgg tcctcagaag ggtgtgtgag98161cctggattgc gccaggagct tgatggtttg caaggacccg tgaatatctg actggggtcg98221acaggtcagg gtaggctgag aatctgcatc cctgaggtcc ccatgacact gaccctacca98281gcacgcggtg cactctagga gcccacatta gatttaagag gaaggggagc aggtgcaaag98341gaatcactcg agctcctcac tcagacggga gcctgggaga ggccgttcct cactcagacg98401agacactggc agaggctgtt tttttttttg ttttttttct ggacagagtc ttgctctatt98461ggccgttgga gtgatctcgg ctcactggaa actccaccte ctgggttcaa ctgattctcc98521tgcctcggcc tccccagtag ttgggattac aggcgcccac caccacgaga actcggttct98581tttctccctg ctcacacgag aggctgccca caccgcttct ctcaagcaag ccagggtgtt98641taggagatga ccttgagcag gtcccgaagt gcatctgggc gggcacaccc catctcacca98701cccacacgta ctcgctggca aactgccggc cttcttgtag cttctgcagt tcccggggca98761gcgggctctg gcagtgtctc ctccaccggc agagcagcac aggctccagg ctcggccacc98821acacctatgg agagagcacc agcacacaga tgagggtggc tgagatggac acacctccgc98881tgccccagcc Rtgacaagcc cccagtgctc ctacaggcca ctacaggccc acactcgccc98941tgcgctctgg aactccaaag ccagtatttt ttacgtcaat taaggctcaa agcagttcca99001tgaggcaYtt attacccaca ctcatggaag acgtggaatt tggagaagga agagcctggc99061ctaacgcaac agtgaaggga ccacactccc ctgagctggg aacgaaacag tgaagggacc99121acagtcccct gagctgggaa tgaaacagtg aagggaccac actcctctga gctgggaaca99181aaacagtgaa gggaccacac tcctctgagc tgggaacaaa acagtgaagg gaccacactc99241ctctgagctg


Following is a FPGT genomic nucleotide sequence (SEQ ID NO: 5).

>1:74033451-741308501gagatcttct atcaggctat cctggggaac tgctccaggc tgaaacaaac caccagttag61gacgaaccgc ctttgatgtg aaagctggga agctagaatg tagtttcgcc ttatccttag121cgggtcggtt ctaacaattt tgtcacccaa actgaaccac tcaccgggtc aactgggctg181caagccctta tgagttggag acaagKggcc cagccccaac acagtcagac aaatcgctgg241gcggccatct gctcctgaga ccgttgctag agaaacaaga caacatccaa gttctccaSa301tcatggtttt ccgggcgcca gccaaggcgc tccgggggcg tggttacgtg ggcgacgcac361ggagggggcg ggtcagaaac aaccgggcgg aggcgcaccc cagggcgcat gcgtgctgtg421cggcgcggtc tcagggaagg tggggctatg gcagctgcta gggaccctcc ggaagtatcg481ctgcgagaag ccacccagcg aaaattgcgg aggttttccg agctaagagg taccagagaa541ggcaccggag ttctcctggg caatgcagag ggagggtgct tttacgtgcc aggaggtttg601ctggactgtc acttcccgtt tacttctcat ctgYaggcct atgacgctcc ccaggagtag661ctagctaata atccttttga gttttttatt cttttcactg tccctgcttg acatgcttgt721tgcttacttt attcttagta gtatagatat taaaaatgag ggcggaggaa gaagaagaaa781atcttactag tgaagttatg cattgattag ctcattaata aaagttgctt taaataaatc841taaaagatac gaagcacagt attttgaact ttatgagtag atgccagccc cctatctact901tcacgcttta cattacgttc atctacttct gtaatgtaac cgtttgttga gggagtaaca961gaacatggag gaattatgga ttaaaaattN aaaggaagaa atttgttatc aaaaaattgg1021ctatgacact taatccctgt acacctgtct aataaaagtg attatatttg tgttgcagtg1081actgcttaaa aaatactggg actaattctt ggataacagt aaaactttca ggactattag1141attctagctg acaagttctg tttgttaact aattaaaatg ctaaatcatt ttgtttatca1201ataaagagat tccaactgtc tactaaaagt gcacactgtc ttttaaagat gtcatcagag1261agttgggacc ttctgttatc cagaatttta tgttatattc taacagccta tgtaagtttg1321catggttcat cttaaagtaa tagaaaataa agtacccaca tttataaaca gcaacattga1381gggaagggat ctttgggttg tgtctttctt agtcacctaa caccaagtgt ccStgataca1441taagaattga tttgcaaagc tggagatttg tttatgttta tctcctcttc ttattgaaga1501attgaggcaa tagtctttgg caagttaaaa gtcgctctct taatcaagaa acattatgtc1561attcagaatt tataattaaa ttggattgca aaccataaaa ggagggagtg atctaaaaga1621aaaaaaagga acttgtgaac agtcttcatt ataactattt tgaatcgtat taatgcacca1681ctaattattc cctaactttc aaaagccaaa gatacatttt ggaagatgat aatggatatt1741accttcttta tcatctttct caataggcaa acctgtggca actagaaaat tctgataatt1801tttgttttca tttccttgct ttttccaaat aggcaaactt gtagcacgtg gagaattctg1861ggacatagtt gcaataacag cggctgatga aaaacaggaa cttgcttaca accaacagct1921gtcagaaaag ctgaaaagaa aggagttacc ccttggagtt caatatcacg tttttgtRga1981tcctgctgga gccaaaattg gtacgcgctt tatggtcagt tttataatat aggtcctaag2041cagaataatc attgaagaaa aggtttctgt gacttacagt gtataagctg catatctcta2101cccacagctt tacaaacttt aaaatttgaa cataaattga ctttttttag aaagataaaa2161ggaatagcaa ctgttcttac ccattatgac ttaccttgaa aaacttccag aatgttactt2221aaaagaWaat tttattagtt ttaatgaaat tcacactatt cagataggaa tactcctttg2281gaccttatca gtagaaccct tgaggtcctc tccactccag agttcctcca gcaatttaca2341tgaagaaagc aattagtggg aaaaaatact cttgacctct tagggacata atgcagaatg2401caaaagaaca ctgtcacatg gcttaaatat aatacatata tttgaaaacc ctagtttagg2461tcaactttgc cctttgaccg tccattgcaa atagaaataa atgctgtttt tctgaatgct2521cttttctatg tgatacttaa agttgaagcc ccctgatacc agagataaga tatctgtata2581tgacattaat atgacattag cattattcta attagccaac attattttaa tccttacaac2641tatgaggtca gtactgttat tatttctttt tttttttttt tttttttttt tgagaaggag2701tcttgctctg ttgcccaggc tggagtgcag tggtgcgatc ttagctcact ccaagctccg2761cctcccaggt tcaggccatt ctcctgcctc agcctcccga gtagctggga ctacaggtga2821ctgccaccag cctggctaat tttgtttttg tattttttag tagagacggg gtttcaccat2881gttagccagg acggtctgta tctcctgacc tcgtgatcca cctgcctcgg cctcccaaag2941tgctgggatt acaggcgtga tattatttcc atagatgaga atctgaggta cYcaagagtt3001aagtaactgg cccaagataa ctagtgtggt agttatattt aagatttttt tttgggaatc3061caagttctaa aaatcttttt tcgaaaggaa ctctaacttg tgtttttgat tgtgtcattt3121gggagaatta tttaaatggg ctttgtgatt tgtaactctg tatgaatgtt tatatttatc3181agtcttctag tgttacagat tgtgtaattg attcttcttt cttataatta ctattttggg3241gagaatatta cctcatcatt tattactgag ggttgttgtc tgggtttcat tgctaattta3301ttgattggtt gtttgctatg agatgttttt aattttgttc tattgccaat acctttgatg3361tccatctatt tacttattcc tgaaaggcaa atccttaagg ttatataaca cctaaattta3421ctgtcatgac aaagttttag taggtttcta ttaaataaat tgtgcttttt taactttgtt3481tttaaggaaa tggaggatca acactttgtg cccttcaatg tttggaaaag ctatatggag3541ataaatggaa ttcttttacc atcttattaa ttcactctgg taatgttata ctttactaat3601ttacattttc tttttagtct tccaccttta atactttaga gacttttttc tttcttgcaa3661acactttcct tcttttttaa aagaatcttt gaagttattc tgctttgaaa acaaatattt3721tatgattttc atgacttttt ttaggttgta agtagtatag tgattttcac aaattttact3781cgtaaaattg tagatcacgt atatcgtcag tcttttatga atattgtttt gttttgggta3841tgtatataga aaattatttt cctaagacaa ctcattcaaa tatgaatagt tctcgggaat3901actttaatgg ggatttttct tttaatttta atttgtattt taagttccag ggtccatgtg3961caggtttgtt acataggtaa acgtgtgcca tggttggttg ctgcacctgt caacccatca4021cctaggtatt aagccagcat gcattagcta tttttcctaa cactctttct cccttaatgg4081ggattcttgc tttgagggaa gggtggttta acactgtcat tactagactt ttgccagtac4141tttttatctt gcatttttgt gtgattattg Rtagccagag gccacaatgc ttaaaaataa4201aaggaaaatg tgacaagata cagagtggac aagtaaatat ataaaagaga aatcacacac4261acaaaataag acattttaaa acagaaaagt aggaaggaca tggtctcaga atRaagaata4321gctttttaaa ttgaatacat ttagttgtgt gaaaattttg gttacatgaa aacctttgtt4381gtttttctat gtatgttaag gggctaatga aaactgtcat gaactgacac cagtccaggg4441acaggtattt tagaattatt catctacatg gcttccaagg cactttttca ttcttatgat4501ttacatacaa aacaatgatt ctttgtaaag atgatgtcaa agtaaagcat ccaagttctg4561gtttgaagcc taaaaatgtg taaaaMaatc taatttacaa ttttatttaa gaaaataatt4621attatcagtt tagtaaaaat taaattagat accaggaaat gcatagttag ataatttttt4681tatttttatt ttttggaaat agataattct tagtgctaaa cttgactatt ggctgctctt4741tgaaaaaaga aaaaaacaga aacttattag ctctcttttg ttgaacaatg tactgtgtgc4801tgccaagtgc tttacaggta ttattgatct tcagtaataa attaacctta gcttactata4861acatttttac tttataaact tcttaatttt tttgactctc ttgtaataac attcagctta4921aaacacaaac actgtacagc tgtacaaaaa tattttcttt atattgttag ggtttatttt4981gttgttttta tttttacttt ttaaatattt tggttagaaa tgaagacaca aacatattag5041cttaggccta cactgggtca ggatcatcag tatcactacc tctatagtcc tactaaagtt5101caggagctgt catctcctat gataacaatg ctgccttctg aaagacctcc tgaaggacct5161gcctgaggcc attttatgct taactttttt ttgtatgtat catatgaatt tattctaaaa5221taacaatgaa aagtatagta tagtaaatac ataaaccagt aaaatagtca tttattacca5281attattatat actatataca taattgtatg ttctgtagtt tagtaaactg gcagcacagc5341aggtttgttt ataccagcat caccacttga gtggtgcatc Rtgctaggac tttacaatgg5401ctccaacatc actaggtgat aggaattttt cagttccatt atgatcttat gggacaacct5461atgtatatgc tgtccataat tgagYgaaat gttgtaaaga agcaagtatt ttagcattag5521tatattagtt ttctataact tttattttct gtgaagtaga tttttttttt tttttgagat5581ggagtctggc tctgtcgccc aggctggagt gcagtggcac gatctccgct cactgctgca5641agcttcgcct cccgggttca cgccattttc ctgcctcagc ctccagagta gctgggacca5701caggcgcccg ccaccacgcc cagctaattt tttgtatttt tagtagagat ggggtttcac5761cgtgttagcc aggatggtct ggatctcctg aYctcgtgat ctgcctgcct cggcctccca5821aagtgttggt attacaggct tgagccaccg cgcctggcct tctgtgaagt agattcttaa5881ttagaagtct aaacatggtt gaaatttgtt gtttaaaatg tagatacagg ctgggtgtgg5941tggctcacgc ctgtaatccc agcactttgg gaggccgagt tgggtgaatc acctgaggtc6001aggagtttga gaccagcctg gccaacatgg tgaaacttcg tctctactaa aaatacaaaa6061attagccagg cgtggtggca ggtgcctgta atcccagcta ctcgggaggc tgaggcagga6121gaatcgcttg aacctgggac acggaggttg tggtgagcca agatcgcacg tcactgcact6181ccagcctggg tgacagtaag acttcatctc aaaaaaaaaa aaacaaatgt agatgcaaca6241tcatgttttg ggccgtggtc cttcttattt gttatacatg taattcaaaa catttcacag6301attctacagt atatttaaat tttttcctga cctacagttt ttaaaattgt cttatatcaa6361atggcaacaa gaaaatttct cttcgaattt ttaatYtgga tttcattatt ctttcctgat6421ttgcagtaac aagtggcata tattaaatat cgttaaaKat attttgattt gtaacttatg6481ggttataaaa ccctggggtt ttgaaaattt tataaataat actgcatttt taaatggtta6541tttcgtttta attttatagg tggctacagt caacgacttc caaatgcaag tgctctggga6601aaaattttca ctgctttacc tcttggtaac cccatttatc agatgctaga attaaaacta6661gccatgtaca ttgatttccc cttaaatatg aatcctggaa ttctggttac ctgtgcagat6721gatattgaac tttatagtat tggagaattt gagtttatta ggtttgacaa acctggcttt6781actgctttag ctcatccttc tagtttgacg ataggtacca cacatggagt atttgtctta6841gatccttttg atgatttaaa acatagagac cttgaataca ggtcttgcca tcgtttcctt6901cataagccca gcatagaaaa gatgtatcag tttaatgctg tgtgtagacc tggaaatttt6961tgtcaacagg actttgctgg gggtgacatt gccgatctta aattagactc tgactatgtc7021tacacagata gcctatttta tatggatcat aaatcagcaa aaatgttact tgctttttat7081gaaaaaatag gcacactgag ctgtgaaata gatgcctatg gtgactttct gcaggctttg7141ggacctggag caactgtgga gtacaccaga aacacatcaa atgtcattaa agaagagtca7201gagttggtag aaatgaggca gagaatattt catcttctta aaggaacatc actaaatgtt7261gttgttctta ataactccaa attttatcac attggaacaa ccgaagaata tttgttttac7321tttacctcag ataacagttt aaagtcagag ctcggcttac agtccataac ttttagtatc7381tttccagata taccagaatg ctctggcaaa acatcctgta tcattcaaag catactggat7441tcaagatgtt ctgtggcacc tggctcagtt gtggagtatt ccagattggg gcctgatgtt7501tcagttgggg aaaactgcat tattagtggt tcttacatcc taacaaaagc tgccctcccc7561gcacattctt ttgtatgttc cttaagctta aagatgaata gatgcttaaa gtatgcaact7621atggcatttg gagtgcaaga caacttgaaa aagagtgtga aaacattgtc agatataaag7681ttacttcaat tctttggagt ctgtttcctg tcatgcttag atgtttggaa tcttaaagtt7741acagaggaac tgttctctgg taacaagaca tgtctgagtt tgtggactgc acgcattttc7801ccagtttgtt cttctttgag tgactcagtt ataacatccc taaagatgtt aaatgctgtt7861aagaacaagt cagcattcag cctgaatagc tataagttgc tgtccattga agaaatgctt7921atctacaaag atgtagaaga tatgataact tacagggaac aaatttttct agaaatcagt7981ttaaaaagca gtttgatgta gagRtatttt aaatattgta cactttgcct ttttgagtaa8041cattccagag ataggtattt tttgtaggct gtttcactga actcagttaa tgaaaactgt8101attaacataa ttgttgtagc ataatattaa tagtgcaaaa gtacatataa gtcattttga8161tgaaaaatat tccaagacta agttgagaaa agagatacta ttttggatgt gtatcagtat8221ttttgttttt aataatgatt gatttgtgga gcattgtttt ttcacataat tagttttaaa8281ggtaattttc taagcatacc tttggaattt ttccatcttt tttgaggctt ttggtccagt8341gaagttctaa gtattcactg gcacttctct cctcaactgt aattctattt ttaataataa8401aaatggcata ctgtagggtc ttcagagtag tgtaggaata ctgtagaaat actttttcag8461aaacgaatcc atagctgaca aattcactca gtgcccaata tattgtgatt attttcgttg8521ataaagaact agatacaaag acctctgaaa ttgatgataa aatttgtatc tcattaattt8581tatcaaaatg aaactaaaag tacatatgta ttatacactt gaacatgtgt ttgtatatct8641ttaaaatttt cctttatgtt ccattccata ggaaaacaca catatgcaca caaaactcag8701ttatctgtgg ggaaagtggt agtaataatt gagattcatc aataatcata taatttcact8761atagacatta cttgcattat ctcctatcag tccttctaac aaaacttMaa ttaccagatg8821aactgacttt agttttcatc ttattttttg tccacatgct ggtgatgctg agaaacaata8881attggcccaa atgcagacat caagaagcag gcaggaaaat ggaaaaactg agataataca8941caggtgataa acagtttcRa gaaaataaaa attggctaga atctcaagtt atgtgtttct9001atattggtat aagcatataa gtgaaaagtc ttataagtgt aatagagaaa agatttgtca9061gtgtgttttt ttaaatgaaa taactagtct gtgctacttt atgtcaatat aaaaattggt9121aaacWagaag taacttgtcc acaaccctca gttatgatac ttatgtgcgt gttttttttt9181ccaaagtttt tcccaaggag aagatacaaa tatatggtag ctattgttta aaaatgattg9241atttacttgc agatttttca Raggatgtta tgtctttgtc attcattaaa tgttcaaaat9301tgtagtttta ccaaatctga agtgccatca gttataagaa tcaccattat tttatgttcc9361attaagaaaa acacaataaa acatgacact gctttttttg ttacttgaaa ttttcatatt9421tattggagag ctcttttaga tgtttttaaa tacagatttt tatttaaccc tcttgcatat9481acatataaaa taacaaaaaa ataaaatatt gagatatcat caagaaattt ctacagcttc9541acattcaaac tcttcagcac tttcagagct attgatatgc atatgtccac ctagtaccat9601cttctctgct ttcagaatta ttggtaaggc tgcatttcct gagtgctcta ccacacttat9661gtctggattt tttttaccca gctacagacc cacactcctg caagatttga tacccttcta9721tttggcaaaa ctcagtttca cttttctttc agtttaatca caggtcctta gaagatcgga9781cctgtgattc tattcaaagt cagaaagaga tttctcacaa attgaYgtca tattaacatg9841tcaatatgaa aattaattgt ttggtaaacc agaagtgacc tgtctgaagc cctcaattgt9901ggtatttgtg tgtttgtttt ttctttttca agattctttt taaagaaaat atatggtagc9961tgttgtttaa catttatcta cttacagagc gttcagagga tgttatatgt ttattaagtg10021ctcaatactg tattttaacc taatctgaat tttaatcata atcctgYaca atgcatggat10081tctggttgct ttgaaatggt tctgttcaaa ggacttatca atgtctcttg ctttgagatg10141tattgcttgg tgtgtagtag cacaaacgta ccaccaataa tgtacctaat tcaactgaag10201tgacagtcat atacgcagat atgaccaagt tcacatgtga gcaagtaaca actccatgac10261tRttgctgcc tggctaatag ctaYtatact atcatttgta aggcacattc tgttttctgg10321ggtgtcgaaa tgtggataaa aaaagatgaa ttaggaatga ttctaaccta agtatgtgtt10381ttaataaaag gatctaggca aaaaaaaaaa aagtaagtca ctcctaaatt taatccctga10441ataatgcagt gtacatgatg atagaggaga agtgtgtttt tgataagaag tgactaaatt10501ttgtgacaat cttaattttt taggcttttc atttagtctt ccaataaaga taattttttt10561ctgagtttta tcagcagtag aaaaataaga cctacatgga taagtttgaa attaaacatt10621ttgtctcatt ttataatttt tatttttatg aaatcatggc Rcttacatca tatactcagg10681tgtgtaaaag cctgctattt gtaactataa tccataaact tattgatgat ttaataacta10741tcaacaaggg catgggaaaa attcaacatt tgataataat cttttacctt aggcatttat10801tctctaagca aagcagaaat aaaggatatg tatcagtgta tactgattga aataaagctt10861taaaaatcca agtgatactc tgaaattgtc aaaagtttaa tggcatgaat ccaggattaa10921ttcccataac ctgtgatgat ctagtcatct tctgtagcta catagtactc ctaatgttaa10981gattaccaaa ttcagcaata aagccaaaat caaaactgcc ttctgtcaca tatagctgtt11041aagtaaacta cacaaaaKca gagttgctgt ttacatacat catgatgccg tctggacttg11101agcaccttgg cggctcagga ttaaggcacc tcagtctagc ctagcagagt tacaatggag11161agaagtgaaa actcattagt agctcaaaac aagctaaagg atttcagatg ctctttttct11221tgttcatcta aatcaagtct tcatctgtgt caagaactaa attgttaaag acctttttaa11281tttgcttagt gcaagaaaaa taatagcaac catatatcta gttaacttga gtcttctgtg11341cctctaaaat tttgtttaga gaaagcctat ttctattgag gttgaggaac acaattctaa11401tacttagtaa aattattaac attcattgca tttaactttg tcgtattttc cttcctccaa11461tactagtgtg ccaaattctg taacatagta gcatttttca tgctcccatt tatctgtata11521tacagaaact tctgcttagg ttttttttta tatataaaga aagtctagtc ataccagttt11581ggttgaagaa tactaattct gtccctggaa gagctgtcta cgtgagccac aacaaaccaa11641acttgccaca tcattcaaac ttgccaattt ggccgtagtt tagtagatga caactttggg11701tcaggttaat atttggggaa atggaataat aaaagctgct gattttccta ctgtattgct11761acttggagga ttggaggtgt agagggaaga tgcctgtaaa tgagtcactg ttatcaccta11821cctgcacatc agcttgattg ttgacactta tgtctaaaga tgttcatccc Ragagtatta11881ttgtgcatgc aagtaactga ttgatactaa cgagttaaca ttttattttg ggctgcttgg11941acaaaaagtc ttatcttttt gaagcctctg gctgcaggga tctcttagcc gttgcacata12001taaatttcaa ccattcacac agagcaattc atttctgatt gctgtagact agtgcaatcc12061atctgctagt gtcatttcct agctatctgt gccagaatta cactccttca attgtgcttc12121agtgcatttc tgaaacaatc taaactaaaa tgtctgtatt ctttggccac tggcttatga12181tgtgttactg atatgcttgt tgaatctcca gcctctttaa cactttttaa aatctttgtt12241acaaccaaca catcatagct aagtaatctg cacatgtaaa tgattttttt Waatttcttt12301tcttaccagt atccactgtt ttctctctgt agcacattca gatggttcaa gtgagtttta12361atttaccact tccttgtagg agagtgtcac atcgcccaga atagaattaa agcatttaca12421tcttgaggca aaaaaatctt ttaaaataca actatgtaga aatcaagtaa gtgttcattt12481catttgcagg tttagaaatg ttgcactgct taatgaagtt ttacgtgagt ggtttcttac12541ctattatttt tagccttggg gtttgagaaa gtaaattatg tccatcaatt ttctgtactt12601tagagaagtt tcaaatattt ctagaactag aaaacttggg actacacaaa gaaataatta12661tgattatatc aggtggccca agtgcagatt ttaaatgtac aggcacactg acaaagtcat12721aaattaaaaa tgcactaatt atatatcact gggatcaatc tttaatgaat tgaataagaa12781agatcactta tgactggtta aagttgatag agcttctaat taagcattat ccagtcaaat12841aaagagctgt gtatggaagc cagcctatga ggaaggaagg aaaaagtttt agagtaattc12901ttttttactt gttgcaacag ctgattcttc aatacaaaag aaatatttac tttgatttgc12961atttctctga tggccagtga tggtgagcat tttttcatgt ttttttggct gcataaatgt13021cttcttttga gaagtgtctg ttcatgtcct tcgcccactt tttgatgggg ttgtttgttt13081ttttcttgta aatttgtttg agttcattgt agattctgga tattagccct tcgtcagatg13141agtaggttgY gaaaattttc tcccattttg taggttgcct gttcactctg atggtagttt13201cttttgctgt gcagaagctc tttaattaga tcccatttgt caattttggc ttttgttgcc13261attgcttttg gtgttttaga catgaagtcc ttgcccatgc ctatgtcctg aatggtaatg13321cctaggtttt cttctagggt ttttatggtt ttaggtctag cgtttaagtc tttagtccat13381cttgaattaa tttttgtata aggtgtaagg aagggatcca gtttcagctt tctacatgtg13441gctagccagt tagaggatgt ggagaaatag gaacactttt acaccgttgg tgggactgta13501aactaattca accattgtgg aagtcagtgt ggcaattcct cagggatcta gaactagaaa13561taccatttga cccagccatc ccattactgg gtatataccc aaaggactat aaatcatgct13621gctataaaga cacatgcaca cgtacgttta ttggggctct attcacaata gcaaagactt13681ggaaccaacc caaatgtcca acaatgatag gctggattaa gaaaatgtgg cacatataca13741ccatggaata ctatgcagcc ataaaaaatg atgagttcac gtcctttgta gggacatgga13801tgaaattgga aatcatcatt ctcagtaaac tatcacaaga acaaaaaacc aaacactgca13861tattctcact cataggtggg aactgaacaa tgagaacaca tggacacagg aaggggaaca13921tcacactctg gggactgttg tggggtgggg ggagggggga gggttagcat taggagatat13981acctaatgct aaatgacgag ttaatgggta aagcacacca gcatggcaca tgtatacata14041tgtaactaac ctgcacgttg tgcacatgta ccctaaaact taaagtataa taataataaa14101ataaaaaaat taaatattta ccatgagttc agtgagaaga agcaatttac aaaattagcc14161atgtttctag tgtctttgag tatgtatctc taagactaca aacgtgctgg aattttccat14221aacataaaaa taaacaaaaa caaaaaacaa tacttctctt agccatatta tttcatcagg14281aatagtcagc ctagtttcat ctacaatgaa accgaagttt tactagccat catacccaac14341actgaattct agattcagat atgtacctcc ctgcttcgta tccctccctt actggaaaaa14401aatacatagg catctctatt tggagagctc taatcttcta tcaattaagg taatcttttt14461catctagata aatgacacag ccattcacca aattgctcaa ggaacaaacc aggaaatttt14521ccatgactct tcaggtactt cccctcatcc ttttcatcct gttttcaaca agtgaagtct14581atgtatgaaa aacacaacca aaatccatca acctctctca acttgttgag ctaccactgt14641tttctagact aatggaatag tttttttttt ttttttacat cccacaatgc cttattgtgt14701gcagtaataa caatgatMtg acttttataa ttgcacatga acgcaaagtt gcagaaagcc14761tgcaaaggaR agatttagaa gattggcatt ggggtgtatg aggcattgaa agaaaaagga14821atactgtggc catctcaacc ttacttcact aatttggtac tgtttttgta tcccattttg14881ctcccctgaa gactaccttt taggacaaag attcatacta ctttctaaca ttctacactt14941tattttctcc atgaagtttt gcctggtgat agtctgcaat ctctttacca cccacattgt15001tgttttttac ctttttcata tttgctatgt gtaatcagat gtcagctgga aatctgcttt15061ccctttagat gtaggcaaca ttagttgagc ctctattcta aaccccagag atgatgtcaa15121aaacttctct caaatatctg tgaaattcct gaagatcata tatctccata attgctttgg15181actgtaagct tccatttctg gtgagttagt tctgtttggg cttgcatgtg attttctcac15241aattctatga gctgagcgcc tatttttcac tatgtgagtc tcaaactttt cctctttcct15301gcaattaggg catcaccata tcctctccag aaaaaggcct tctcacacaa attttgtgtg15361tagcaaattt ctctgaatca ttggagctag ggaggaagRt gctggaagga aagagaagga15421tacatatcag agcacagcat taagtaataa ttgttcttag aaatcctttg gcatcttcat15481ctccaagact ttcttcacta gctatttttt ttataatcca aaattatgtc taactttttt15541tctcttcctc agaccaactt cttccatccc cttacgagat atgcacacgt tgtttacatt15601gtagacatat agcatttatc tcttctttct cttgtgtcat tagttttctt ccttcaatga15661attattctca tcagtacact aatattctgt gatttttcca gtttaaaaaa aaaaaaccct15721tcactacact tccttcaggt attactttat ttccttgttc cttttacact aaaattttta15781gaatacattg tctgtttttc cattcttcag ttcccgtctt cctatttact cttcaaccca15841attcaggctt ttagcacccc tctccattac cactactcca ttaaacttgt tctgaagatt15901tccagtgctc tcgactttgc taaatccagg ggtcattctt ggtctcatct tcttgccctt15961taaacagcat ttatctcaat tgattacttc tttgacttgc attctttacc tggatgcaag16021gaaccccaca ttactggttc tttttctaac tttctggtca cactttttaa gtctctttaa16081ctgatttctc ctcatcttcc tcacctcata tcactagaag gtcacagata tttgtRtttg16141aatttctcta tccacattca ctcctttgga ataccatcca ttttcatagc atctatgtgt16201tatcaattct caaatctgta tttctcatta tcaaaatgta actctcaaat tgttaaaata16261tgaatccata aattgtgaac atgggtcaga atttaatcag tgaggaagcc agcaggatta16321tgaagttgat ttttatattg ttccRgatgc tacaaggaag aaaatttata tacaacatta16381taaagaattt tttccaataa cagagtacat tttcttatga gataataggc ttaaaagaca16441atttagcaag agttttctca agaagttctt attttagata agaggtagga ttgaggcagg16501aaaatagggc ctggaggcag ggaacgtaag gccaattcac acttaagcta tgacaggaaa16561tatccttttc atagggtgta tgcagagtaa ataactttgt aactttactt cttcctctcc16621atttacatat ggcatacacc aagtaaccag tggaaatctc tagagggtat ttaaaccccc16681acaaattctg taacggtgtc cttgagcccc tatgctcagg cccaYtccca cactgtagag16741tgtactttca ttttgaataa agttcttcat tctttctttg ctttgtgtgt tttgtccagt16801tctttgttca aaatgccaag aacctggtca ctctccacca gtaacaggat tagataatga16861ctgtgggttg gatttcctga aaaccagaat ctgagatgta ctttagaatg caaaatattt16921ttaaggaggg cctgtgggat taatacttgt acaaggaagg ggagagagca aaacagagta16981gaagaaagtg aagctgagat gcaatcccaa agacagcctt aattggctaa tcccaagagg17041agctctggat attagaatag cacttcagtg tttcccaagt tgaacctgat tggccaggat17101ttataccaca catcaattat cttttctcag gtgcccaatt gataaggaag acaatggaca17161agatatctct ttaggaataa gccaatctgt gaaagggggc tgaccaatga aagctgtcta17221ctgacagcac ttgcagtagc tgaggtaaca agcccttcac tgaagaagca atatgggtgc17281catcacagtg tccaccacaa taacattata aagtccactc aattccaaga ttcaatttct17341atatgtaact attttaatga aacattgtta tgttttaatt caagttgaat tatgacttcc17401aaactaatca tgcaatattg tacatatttt aagtgtaccc aatccaagaa gaaactatac17461tttagtagat atgtcaatca ttgatttaca gaagcttttt ttttgtaata ctgtcacaaa17521tctacatgat aatccctagt gccagatttt acttttgact agctgaRcaa gctaaaatat17581tcataaattc acatgaagcc agatggcttc ccactgaata tgcatttgaa tgattcctgt17641acattattta ttccaagaag tgcacaaaat ataccatcgt catagtagag attcYtaata17701agtttgtttc aaaaaggagt caaggaataa caggtgtcca gtgctaacta tattttggac17761tgaaccatga ttctatgtat atttgagaat tactaacagc agattcaata tagttagttt17821ctattagtct ctgaaatatc aggaatctga aataaggaag gttcagcaag aatgactagc17881attttggaaa aaagaggatt taatgtggct agcataattt cttaataatt tgcttagttc17941tctaaaatat tcaaatgtac aacacaaagt attaaccatt tttgtcaagt ctatcagtat18001gtttttaggg gtactttttt caggttaaaa taacagatgt gaatttgaaa tgaaaatata18061acttaatcat taaaagcttt ttctccaaca tatttgggga gagaataaat gaaaatagta18121tttttttctt ttttaattca tagagttttc ctttcattta aatctgctgt gtcccttttt18181aaagtttcct gttctctgga catctgtcat cattgctgtc cccaccttcc atcttctgtt18241tttttccttc aaatgtggta atcaatgttg ttttgtaaaa caaatctaac aattctacta18301tctgaagtca ggatggatat gcttctgatc cttcttcagt gttttgttgc tatgctttca18361tgtgtatctg attgactttg tttgttggtc attgtcctta gcaaagtatt tgcagaggtt18421tcccatggcc taagatgaat ggactctcct gtRgaggctt tgcatttgct tctatcagga18481gaccagagaa gcattgccac ttaagagcaa cctcagatta agttcaagag tgagcttttc18541tgacttagaa ggctctgtat aaatccaaag tgaatatttt tgtagcaata attgctcagg18601aaccttattt tactttcttc tcttttttcc tttcttcttc tttttttaaa tttcttctgc18661tctgcttaac agcaaggcaa gctgcttgac aattctttgg tttgggataa aggaggacat18721atttacttct agctgatWct tatMcaaagg catatttttg ggaggcttta gattattagg18781gtgtcaaaga caccctctaa gaatttccga cctgaggaaa cacttggcta ccctctggtt18841ccttagaacc taagtttaaa tgtgtgcttt tggaaaattc ctcagggcag aaacttactc18901catacgacct acatttctta tacaaagaaa tatccactta tatctctgtg tttaagcatt18961ttcctgaaat ggtttgtgta tacaacttta tggtttttag ttcctcattg ttttttgcga19021ttatgacttt tatacttagc atttaaaatc agtccattat tctcacttat gtgaaacata19081actctagttc tgtattatct tattgactta ttatccaaca ttttcactag tgatatattt19141tagttagtag agggtatggt atgttagtgg gggggatagt actagtagat gatatatttt19201tataatgaat gttacagtca tggaatgtaa tgattatgaa aatgctaaac atatgaagaa19261aaaatgagga ataaagtaaa tatttatacc cttatccctt ttatccccca tcccataagc19321cacatgacgg agtagacagc tcagcaccta gcatgatgct gctatgagat gataaaccat19381gttaaatctt ttacagagca ggtcagaata aacataagca gacaaaagta tgttaaatgt19441atactttttg aaataggaaa taagcctaca aaatttgctt taaaaaagtt accatttata19501gtagatagaa cacatagacc agtgttagac cccctctgac ttcaaattat cacaatttga19561agttaattat gtgccaggat atttgatcag cactaagaat attttatccc ttgctctcag19621ggttcttcca gtttagttga ggataagata ggaaaaaaaa taattgcaat aattcctagg19681tagagctacc ataagtaagg catttgaata gtcagccaaa gagactaaca aaagctacat19741gatgaggaaa gatctcttag tttggtacgc gagagtgctc ggtgcataRc aaagtctgaa19801agaaacccag tcaaagtgta gtgtgtcact tcagaaatca catggaagtg taagagtggc19861ttgggaaaag gtctacactt gattaattta tagtatttta tataaatatg cagataattt19921catatttata taaatacaca ggtaccttag aaattaaacc tctgaccaaa ttatgattaa19981caatttattt caaagaagag gaaagaagag agaaagtcgt atgtgtttat gtgtgtgtat20041gtaatatggg agtgatattc ccagtatttg gctaccaaat gacaaaattg agtttagaca20101ttgatgaata gtaatgaata aggaaacaaa taataataat aattaatgtc ttcccacaga20161tcacttccta ataaaacact gtaaattaaa ggatatggta atttatgtga tagctttgct20221taacgataag cagcacttat tgaactgtgt agaactgaaa caaaagagaa ctggaaaaga20281gaaacttcac aaatgaatta acatagtaca aatattgtac agcagtttat ttactttgaa20341cctgaaaaaa tttaattaga tttcaaaacc ttataaaaac cttattttat Rttctcgtta20401aaacacatat ttcttggaaa tgattttgtt tagacatgag taggtacttt tgtctttaca20461aaagtttatc cttatctgtg ctgagctaaa aattcagtat acagctggtg ttttgattga20521tacagtatag aagtgacacc acatattatt atatcagtct gatttaaaga gctgaaagaa20581ggtgtttatg gagtaaacta cactgcataa actttttttt catggctctt agtagaaaaa20641taacaacagc aacgaccaaa atatgtacac aaatgcacat tttatatttc atagtcagac20701attaactaaa ccaataaaac aatcttcaaa gatttttatt gcataaaaac aaaatataac20761tttctagtta agcattaagc tcaacaaagg aaatgtataa aaatgaaaaa acattttagc20821tttgttccta gcactgctat taactcaccg tgatattcat taaaccaaaa gtcaaatata20881tgaacagcaa gggccactgg gcattgcaag tctgaaatgg tgtgctagaa agtgttttga20941aatttaaagg aaaagaaaaa aaaaaacctt cccgacaggt gaaaataagc ctcctattca21001gttatgatat ctaagaaaat ttaatggaat tattaaagga gcacttacag aatatgtgca21061attactttct ttagatggga caaatatatt tcataactga tttcttcatt tccaagaaca21121gttttatttc tttgatcttt ctttcatgat aatgaggtag atgtgtacag gattgaacat21181tacatttcaa atgtaatccc aacagacttg caacaagcaa attttttatg ttattcttta21241taaaatgata cctctgttta aaccactcac tttcttcaac atactatgaa ttcatatata21301atttgtgtat ttctcccact tacagaacac tggcttatac cactcttcag gtacttctct21361cctccaataa ttatattgtg tttttgtggg gttttttttt tttagctttt ctaagttctt21421ttggtttgat tacatgtttc tttttgtaac tctttttatc ttttctgttc atagtttcta21481ccattgctgc tactttatat aatttctctg tcatttgata tttatgactt tgcctaattt21541agcctcatca atttttagtc cccaaaaaaa atccaagatg aaatttaaat tactttggtc21601ccatcaattt tgtttaattt gtttactgtg taaaacaata atagagattt taatgaacat21661catcttaata agttttctag ttctctgatg tgatataatc taaattgtgt ccagactctt21721gattYatcat attttagata ttaaattgtt atttattttc ccactgtcta agatttttaa21781aaatctacaa gtagcaagac ttattgcaaa tcatgccaga ttcaaaaatc cttctgtact21841actagcccga ttaacccttt cttcctcttt catagcatgt ggaagttttt tttttttttt21901tttgatactc atcaaaagct actttgtaaa ataagttagt tagatttttg aatatcaaat21961taggttgtaa gctctgtaag caagggccct gccttatact tccttataga caagctgtgt22021caaaccactt tttggagttt tattttattt ctaattcaag gacctaacct cagcagattc22081tgatttagtt gatcacgagt aggttcttgt aattgtagtt tgtaaaatct ccctgaaagt22141ctggtttgta agttattgct agaacatcaa atcccttgta taaattacta atgctagcaa22201ctccaactaa tgcaaaaacc tcttaaggat tacatataca ggaagatgag gaaaacaaag22261agactaatat ttgtgtgaat tttccttgca tagaatttca aaagcctgta gtatttaggc22321tgcagaattg caatggccta ctctagtttc actttacctc ttttccgacc ttagcaatat22381tcctcttact gaaggaatgt attctaagca attcaattgg ttcagtccct ttcagttcag22441ttcaatttta ttcagttcaa ttcattccaa aacaattaaa tttatgtcta tacttataag22501gaactgtggt gggctatttc aaatgcaaag atgcctgcat tcaagttact tattgcctgg22561gaggagggat aaaagcatta attctatatt ctataataaa tcccttaata gaagcagatg22621ctgcatggat agcaacttag agtaggattt aaatgtgttt aatttcttgc ctattcttaa22681actacatata tgtattgtat atatgtatat atatttatgt aatgtataca ttgtattata22741tatagtatat atgcacacat ataaaattat agtataaact taccaaatat aaatatgttt22801agtcttcaaa atgaaatagt tactaacagc catcttcagt aacattaaag gatgcagccc22861tgatagatca tctcaaggtg actttgtaaa ttagtacatg tccagactct gtagaagaaa22921tgagttttat ataataagtc aaagagagaa tatatagtct ttaggggacc cgaacataac22981tattgtagtt cactaatagg ttaactgtta tatagctgca ccgagcatca tgggcaatga23041gagtttcaga aagataatga tggagttaat ttgtgaaaac tctcatgtga taatatgcat23101ttctctctcg ttctgatatg cttaaggcct acacattttc caaatgccgt gttacccttt23161accataggcc ctatcattta ttctctggct gtatatatga actagggaaa tatgggtgat23221aattttaggt aactatttgt gtactcagac ctagacctca acctgggaaa ctccggccag23281cagttaggaa atgataacac atggtgagcc aacttaaaca aatagactca agacttgtaa23341tcaaatccag tatattctat tatgatttaa taaaccYagg catcagaaac accacagtta23401gaattgtctt tgcatcattt gctttggacc tcaagagggg gtgaccttaa attataaagt23461tactggtacg ggaagaatga gacccccgga caacacRgag gtgcttctaa cacctttgca23521tgtacaattt ccttatgtaa ttcagaacat gcctcttagt attgatgaca atttcatttt23581taaagtttct agcttgagct taagtccatt aattcacaat ttttcttctc cattagtgtg23641aacttcatgt gaccaaatta tgggtttaag agacaaaaaa gggtatgaga tacaacggga23701gcagcaagga caacatggcc aaaatgttct gtgaagttgg taaagactcc tcagaacagc23761tgacattttg aagcaggaat atgactttcc tggagataaa ggcaagaata ctagagcaat23821cactctccat attaccagac aaggaagtac aatttaggct tactagtagc aagaactcct23881aaaaatggct ttctgtgaaa acaagtggtc ttgaaattca taataattat agtctactct23941tgataatcca tgttgaagaa ggggagtaca gttgtaagga aaacatgaag tttttcacaa24001attgtaatta aaatgtaaat ctttttgtat atacttagat actaaacttg aattctggtt24061gggaagggta tattatttgg atgccaataa aatgagatat aaatctatga ggacatatgt24121tgaattggaa gcactcaact tgggtttatt attaaaagtc tgcctggcat taaggatgca24181ctcaaatatt acttgctata acaagtccct aaataattgg cttcagagac aatagtctct24241gcaaaaagaa atcagaaatc tgtgtgacca tacaaactgc aactttctat taattcacca24301aatgaaatgt gttgatccac taatggggtt atgaatgaaa gctgagtttt tttttttttt24361tttaccccaa cagcattata ctaactccta gcactccttt aaattttatg caagatgtta24421aagattatat atgtggaatt tcaagtatga aaagagccag tctagctcca caactaaata24481tacaatgtat agaaacaaaa tacaattgaa tcagagttag ttcttattgg tggagtgctg24541agcttgccaa tacgatgaag gaagatcatg aacctttgcg tacacacaaa ttgctaatgc24601tatttctgcc actgttatag agaatggtga caagggggtt aaaaaacata gtacatgata24661actataatca ccattctggc attaagaaga tataggagaa gatgtaacaa aaataatggt24721aactacagac agagagctta ctgagacttt ctctgcttgg cgggaataag caaagaactg24781gtcctatgga atcactcatc atgctaactc aaagatctta cataaaccaa gaggcatggt24841gcttagatRa tgctcttctg tctcagggcc ttcaactttt tttttttgag acagggtttc24901gctcttgttg cccaggctgg agtgcaatgg cgcgatctcg gttcaggcat ggcatttccc24961ttaaagttga agaactacca ggaccgattt taattttaag tggccacagt gtagcagaca25021cagcagcttc ctgctttctc tatcactgga gtctgctgca gtaagcaaca gggaattagt25081cactgagtca acaaggggga accacccaaa cttgtttatt taattttgct acataatagc25141tacatatcta tggattctct aacaaccaca ttatatagtc ccacttttca aatctttaaa25201taatttgtcc tgcctatatc aaaatgtatt tcttttcact ttgaggaaca gccacaggta25261gataattgac aagtgaaaag aaaactctca aaaagaagag gaaagagtaa agaggacttg25321gtcatccatc ttcgatacca gctcagtcac ggtagaatat ggcaccagtc agagtcatga25381ggtgcccctt ctaggctcta gcttcaggac aacatttcta aacataccct gggccagaaa25441cgaacctgct gccctgaagg gaaggaccaa gttttggcag gacccatcac ctgctgacta25501aacagccttt gggccctgaa taaccagcag tgatacttag gttgtatgct gtgcactttg25561ggtgagactc tgagacttgc tggattcaga tgagactcag cacttttcca gctgtggtag25621ctacagggag agacaccttc tacttgagaa aagcagagag aaaataaagg ggactttgtc25681ttgcagctta ggtatcagct cagccacagt caggaagagc accaagctct taaggtccct25741aattccaggc cttggctctt ggacagcgtt ttgagacctg ccctgggcta gaggtgagac25801cattgccctg aaagttgagt cccgggccag gtggcattca ccacaagctg acttaacagc25861ccttgggtct taaaggaaca ttggcagtag tctggctgta ctcctcatgg gccagtgatg25921gtggtagtca ctgtgtgacg ttcctctgcc tatggaaagg ggagggaaga gtggaaaaga25981ctgtgtctca ttgtttgggt gccagctcag ccaaagtgca ataaaacacc aggtagactt26041ccaaggtttt gactttagta cctggctcca agatggcacc tctggacctg cacaggacct26101ggagaatctt gccatcctga aaagtaggac acaaacctga ttggctttcc cctcctcctg26161ctgattatag agcccagagc cttgagtgaa cataggcggt agccaggtag aggttatagt26221ggatcttgag tgagacccag tgtgaagccc cagtggtggt ggccacaggg atgcttgtgt26281catcccactt ccagtttcag tcagctcagg acagagagaa agattttgtt agtttgtgag26341aaagtaaggg aagagaacaa gagtctctgc ctaaaaattc agagaattat tccagatctt26401atccaagact atcaaggtaa tacctccaca agtctccaac agttatagca ttaataggct26461tggggtgccc ctgaatacag atgtggctta gacaacaaca cccaagtact ttggaatacc26521tggaaagtct tcccaagaag aacaaatacg aacaagccca gacagcaaag attacaataa26581acaccttact cttcaatgcc caaagatgaa tatccacaaa catcaaggct tttcgggaaa26641acatgatgtt gccaaatgaa ctaaataaga cacgagtgac caatcctgca gagacagaga26701tatttaattt ttcagagaat ttaaagtagc tgttttgagg aaactcaaag aaattcaaga26761taacacagag aactcagaat tcactcagag aaatttaaca aagagattga aataattttt26821taaaaatcaa gcagaaatta tggagtttag aaatgcaatt ggcatatatt gaagaatgca26881tgagagtctt aatagcagaa ctggtcaaac agaaaaaaga actaatgagc ttaaagacag26941gctatttgaa aatacacaga ggagacaaaa gaataaaaaa gaatgaagca tgcctacatg27001atctagaaaa tagcctcaaa aggaaaaatc tcggagttaa tggccttaaa gaggaggtag27061agaaggagct agatagtgat aaaatatcca aagggatcat aacagagaac ttcccaaatc27121tagagaaaca catcaatgtc caaatacaag aaatctatag aacaccaagc agatttaacc27181caaagaagac tacctcaagg catttaacaa tcaaacaccc aaagtcaagg ataaagaaaa27241aaatcctaaa agcagcaaga gaaaggaaac aaataacata taatggagct ccagtacatt27301ggctgaagac ttttcagtgg aaaccttaca ggtcaggaga gagtggcata atatatttaa27361agttctgaag aaaaaaaaac tgttaccgta gcatagtata tctggtgaaa atattcttca27421aacttgaagg agaaatactt tctcagacac actaaggctg aggaatttca tcaacaccag27481acctgtccta taataactgc taaaggaagt acttcaatcc gaaataaaag ggagttaacg27541agcagtaaga aatcatctga aagtacaaaa ctcactggta atagtaagta aaaacacaga27601atattataac actctaactg tggtatgtaa accactcaag tagaaagact aaaagacgag27661ccaatcaaaa ataactacaa caacaaattt ttaagacata gatagtacat taagatataa27721atggaaacaa caaaaaatta aaagtggggg cacaagttaa ggtgtagagt ttttatttgt27781tttctctatg cttatttgtt tatgcaaaga gtgttaagtt gttatcagct taaaataatg27841ggttataaga tagtatttgc aagcctcatg gtaacctcac atcaaaaaac atgtggtaga27901tacacaaaaa atagaaagca agaaaacaaa tcataccacc aaagaaaatc accttcactg27961aaagggaggc aggaaggaag atagggagag aaaatcacag aacaactaga aaataaatta28021aaaaatagca ggaactaagt ccctatttat caataataat attgaatgta aatgggataa28081actttcaaat taaaagacat agactggctg aatggatgga aaaaaagact caataatctg28141ttgcctacaa gaaacatact ttgtctataa agatacatat aggctgaaaa taaagggatg28201aaaaaagata ttctatgcca atggaaacca aaaaagagca tgagtagcta tacttatatc28261agaaaaaaat agatttcaat acaaaagcta aaaaaagagt caaagaagtc attacataat28321ggtatagggg tcaattcagc aagaggatat aacaattcta aatatatatg cacccaacac28381tggagcgctc aggtatataa aacaaacttt gttagaagta aagagagaga taggccctaa28441tacaataata gctggagact tcaacacccg acttttcagc actagataga tagtctagac28501aggaaatcaa caaagaaaca ttggactttg tctgcactat gcaccaaata gacgaaatag28561gtatttacag aacatttcgt caaatggctt tggaatacac attttttccc tcagcacttg28621ggtcattctc aaggacagat catatgttag gttacaaaac atgtcttgaa acattcacaa28681aaattgaaat aatatcaggg atcttctctg gccacaatgg aataaaacta gaaatcaaca28741tgagaaattc tggaaactgt acaaacacaa ggaaattaaa caatatgctc ctgaatattc28801agtgggtcaa tgtagaaatt aagaaggaaa ttgaaatatt tcttgaaaca aatgttagtg28861gaaacaaaat ataccaaaat ctttgggata cagcgaaagc agaacaagaa ggaagtttat28921agcgataagt gcctacatca gaaaagtaga aaaacttcaa atacacagcc taatgataca28981ccttaaagaa ctagagaagc aaaaccaagc caaacccaaa attaatagaa aagaaataat29041aaagactaga gcaagaaaaa tgaatttgaa ataaaatgtc aatgaaacaa aaagggtttt29101tcttgaaaag ataaacaaaa tgtacaaacc tttagccagg ctaagaaaaa aagagaaatg29161atctaaataa aaaatgaaaa aggaaacaat acaactgttg tcacagaaat tcagaggatc29221attattggct actatgagca actatatgcc aataaattgg aaaatataga agaagtgaat29281caatttcaag acacatacaa cctaccaaaa tttaaccatg aggaaatcca aaacctaaac29341agacgaataa caagtaaaga gaccaaagca ataataaaaa gcacctccca gcaaagaaaa29401gcctggactc aatggtttca ttgcttaatt ctaccaaaca ttttaaaaaa gaaataatac29461caatcctact ctaactgttt ggaataatgg aaaaggaagg aatacttcca aacttattcc29521atgaggctag tagtaccctg atactaaaac caaagacctg tcaaaaaatg aaaactatat29581gccagtatca ctgatgaata ttgatgcaaa aaccttcaat aaaatacgag caatccaaat29641tcaacaacac attgaaaagg tcaccatcat gaccaagtgg gatttatctt agggatgcaa29701gaatggttca acatatgcaa atcaattaat gtactacatt gtatcaacca aattaagtac29761gaaaatcaca tggacatttc aattggtgct gaaaaaccat ttgctaaaat tcaacatctt29821ttcatgataa aaatcataaa actggatatg gaagaaacat acttaaacac aataaaatcc29881atctatgaca aatccatagc tagtatcatt gtaaatgata ggctttcctc taagatgtgg29941aacacaagga tgcccatttt tcaccactgt tattcaacat agcactggaa ctcctagcta30001gagcaatcag agaagaggga aaaaaggact tcRaaattga aaagaagcaa gtcaaattat30061ccttatttac acatgatatg atcttatatt tggaaaaacc taaagactcc acaagaaaac30121gacttgaatt gataaattta gtaaagttgc aggatacaaa atgaacatac acaaaaatca30181gtagcatttc tctatgccaa cagcaaacaa tctgaaaaag aaatcaagaa attaatccct30241cttatagtag ctacaaataa aattaaatat ctaggaatta atgaaaaaat tgaaagacct30301ctacaataaa aactaaaaaa cattgatgaa ggaaattgaa gacacacaaa aatgaaaaaa30361ttgtccatgt ttgtgaattg aaaaaattaa tattgttaaa atgttcatac tacccaaagc30421aatctacaga ctcaatgcaa tccctatcaa aataccaatg acattattca gtgaaataga30481aaaaaaaatt taaaatttat atggaaccac aaaagaccca gagtagccaa agctatccta30541cgcaaaaaga acaaaactgg aggaatcaca ctacttgact ttaaattatt ctacagagct30601acagcatcca aaatggcatg atactggcat aaaaacagac acatagatga acggaacaga30661atagacaacc cagaggtaaa ttcatacatc tacaatgaac tcatttttga caaggatgcc30721aagcacatac attgggaaaa ggacagtgtc ttcaataaat ggtgctggga aaactcggta30781tctatatgca gaagaaagaa actagacctc tatctcttgc catatagaaa aattacatcc30841aaatggatta aagacttaac tacaagacct caaagtatga cactactaaa ataaaacatt30901ggggaaactc tccaggacat tgaactgggc aaagatttct caagtagtat cccatgagca30961caggcaacca aagcagaaat ggacagatgg gatcacatca agttaaagag ttgctgcata31021gaaaagaaac aatcaacaat gtgaagacaa cacacgaaat gggtgaaaat atttgcaaac31081tacccgtctg atagggattc attaatatcc agaattttat aaagagctca gacaagttta31141caggaaaaac atcaataatc ttatttaaaa atgtgcaaaa tatttgaaaa gatatttctc31201agaagaagac atacaaatga caaacaggca tatgaaaagg tgctcaacat catttataat31261caggaaaatg caaatcaata ctacaatgag atatcttctc acctcagtta aaatggctta31321tatcaaaaag gcaataacaa atgctggcaa gaatgtgcag aaaagggaat tcttgtacac31381tgttagtatg aatgaaaatg aatacaacca ctatggagaa cagcttgtag tttcctcaaa31441aaaataaaga ttgagctatc atataatcca gcaatcccac tgctgaatta catatacaaa31501agaaaagaaa taagtgtatc aaagagataa cttcactcct atgtttgtgg catcattgtt31561cacgatagct aagatttgaa agcaacctaa gtgtccatca atagatgaat ggataaagaa31621aatgtggtac ataaatgcaa tggagtacta ctcagccata aaaaagaatg agatactgta31681atttgcaaca acatagatgg tattggaaat cattatgtga agtgaaataa gccacacaca31741gaaagacaaa ttttgcatgt tctcatttat ttatgggagc taaaaataaa agcagttgaa31801ctcaaggaga tagagaatac aagagtggct accagaagct gggaagggta gcaggtggtg31861gggttgggga tggttaaggg gtacaaaaaa tagttcaaaa gaacgaataa gacctagtgt31921ttgttagcac aacagggtga ctacagtcaa aaataattta attatacatt tgaaaataac31981taaaagtttc taatttgatt gtttgcaaca aaacagataa atgcttgatg ggatggatac32041cctatttgcc ctgatgtgct tattacgcac tgtatgtctc tatcaatata tctcatgtgg32101cccttaaata tatatatatc tgctatatac ccacacaaat taaaattgaa aaattaaaaa32161aaagaaaata aaaattgaga aaagaaaaaa aaactctcaa ataagtagaa ctattatttt32221gttttccctc cccccaattt tttatgtttg tttattatac gatttagaaa cctctaatta32281gagaaacctc taatttttaa aacaaacaaa cataaagttg attacattag attctaaata32341acatttttaa aaatgttatt tgtccaaaac aataattagt tattaattga actataatta32401tcacacaata tcatttttaa gctggttaat taactcttag gatgggtgaa gaaagaatta32461agaaggaacg gaagtggcct gagtcatgaa gtttagatta ttctgattat tttaattcct32521caaagaaact ttttctgatt gaaatttagc ctttaataga tggactttct catgccaatc32581atgtatatct catcatagga caaatactgc attgcggtta cctgtttact catgcatctc32641tggccccatg agctctgtgg atttcttggt acaatacgta tttgttgaat ggataaatgg32701actgatcagt tcgtaaattg agctagactg acaattcaaa gaagtcatat aaatttggaa32761aagaaatttt agccagttac agaaatgtga ctcttacctc tatttaatct agattattta32821atcaacttta tttcatgttt acccaccaac aaaggaaatc acaggaaaaa ctatgatctt32881tgcatttgct gttatacttt attggacaca gaaatagcaa tattgtttat tcctttttaa32941aatatactct gacagttaaa gatactgaga acacatactt aaattactgt gacattatat33001tatcatatta taaatatata ttcattataa tgtattctca agtcttactc atttttgagg33061attaatctga atattacctc aacatgttca gactgatgat ggtgatttat acatcatcca33121acatactaaa aacttagata ttctagtgaa aaagaaagaa aattgtttga taaataagac33181tcttacaaat tatgcttgtt taaatgcaga aacacattat ttaaaagaaa attaatcaaa33241taattgtgtg cacttgaata gttgtatgca tcttaggcaa tggtcaattt atttttaaaa33301taattttatt tcaacacaaa ttatttaatc tttattaaaa ataaaggaaa aaaacctaaa33361tataaactaa ctagtaattc caccttctta tgtttaaagc aatgattaat ttgtcaacca33421gatgcaacgc attgaaactc tatgcttccc caaaacatgg aaatatatta ataaggacta33481atatacaatg aagaaattat caaagtataa attaattcaa cttctaaatg tgataataat33541tgttccaagt gcattagttt gatttcatgg gatacatttt cttttatcat ttctctgttg33601gaaccataaa ggtagaagtg tcaaggtcaa tttttacgtt tttcattttg aaagcatcct33661tttaaatctt tataaagaaa tcagtataaa ataaatataa atagcagatg cgtaagtgta33721gattgggata aataaatcgt ataaacccca aataaaataa tgcagaaaga acataagtgg33781gtttattgaa ggagtattag aatataacta tttgtatgca gaagtggtct gagaaatcag33841ttgaaaagac agaggcatta aaactagatt taggcaattc agtagaatat caaacttagt33901tgaaacaaag tagttgaaga aataatattt cataaatgtg ttctgacatt ttctaatgaa33961gaaacgagca agcataaaag aaagaaacct tactattaat catgtatttg agttggcttt34021tctttataaa ttattaaatt tccttgagta cttcaaatgc tctttttaag gaattattat34081tttttgcatc atttgtttta tatttttcca taatttttta aattaatatt ttgttagaat34141tgttttcagt ttacaaaaaa catggatcag aaagtacaga gttactatat actcccccta34201cacacacaca atttttctta ttaacatctt atatagtata tttgttaaca tgaatgaacc34261aatattgata aatgattttt aactaaagtc catagtttac gttagtgttg tgtagttcta34321tgatttttgc caaatggata atgtcatgta tccaccatta gagtatcata cagaatattt34381ctctactcta aagatttcct gtgcttcacc tactcatctc ttcctctcct ctacccctgt34441aaccactgat ctttttatgg tttgtatagt tttgcctttt ccaaaatgtc atatagttga34501aatcacacag gatgtagcct tatcagactg gcttatttca cttagtgata tacatttaaa34561gttcttccat gaattttcat ggctcctttc ttttcattgc tgaataatct tccatattct34621ggatatacca cagtttgtct ctccattcac ttatgaagga atatcttggt tggttcaatt34681tttgtgaatc acttttaaaa tacgagacta tacttgcata gccaaatcat gtattgcccc34741aggaattacc acaggataga actctttgat aatctgaagt aatctttctt ttgtttttat34801tctacttatc tcttttctct tgcatgaaat tcaaaggtaa aaaaatttct gttaagtagc34861atctcacttt aagcaacctt aaaatagtgc ttcttaaatg atgctccaag tagcatctac34921attagaatca tcagagttgc ttgttaaagt tcagattcct gggctctact cttgatgtac34981tgaatcagaa tttctgggga tggagcccag gtttctattt ttttaaacaa agctcctcat35041gttgcctacc ctcactaaac tttgaaagca gactactttt gtaatcaagt tgtgtttttg35101ataatgtatg atataacatg ctttctccac ttttggacgt ctcaggactg gggaaagtag35161gattcaactt agagtatgtg gccaactctt tattagaaag aagtactttt tacagaaatg35221tactgagtat tagggaaaat attttttgat tctatgttgt ttctttccaa cattctttgg35281aaaaggaatg caaggatatt atcctggacc atttacattt ccagcatttc ttcatcaggc35341tgaaggtacc cagaaaaaat tactctagta cctcaaacca ttacctagat ccacattctt35401aagaagggct tagagataaa tattgtttct ttcaaaccag gaaactgtta tagttcaata35461gtggttaata ttttgggtaa tattttgagt aattttctgc atttcttata ttgctatcgt35521ttcattttat ttccagcaaa atgtattgca ttagaatgtg ggagaattca actgagttgc35581tgaKtggatt ttcaaactca tcgcagaaat cacacctttg aaagccaaag ctttcatttg35641taaaacctag gctttttaag agaataaaag tattcttgaa aatcaaaagg aaaaattcta35701aaaagcagta catcagactt gtgtcccaag caggaaatta gaaacacagt ggaagatgat35761gtaagagcta atttttcctc catatgtgga aaacatacat gtacacacac acacacggat35821ggaagatccg agatattaga aataagaaaa gagatcaaga aaaatatatt tagttgtcct35881ctagaaggct cccatctttg tccttctgga ttcagagtat agaaaccatg attattttaa35941gaaactctga gaggagaaag agctggtgca ttatttccta aagtgtagcc ctgggagatt36001gcaagtcttt taaagaaagc tatccagtat ggtcttcaag ataaggaaga agagattaag36061caggaaggtc cctggacaaa acaaagactt tccaaactga gaaaatgggg gtcacagaag36121tgcttagaga aagctggtgg aactgaagat gtctggtttt caaacaagga gaatgtagag36181tgtatggacc aaggtaacca gatgccactg cagtatagca agtatcaaca gctgtggtag36241gcatgtactg ataagaaggg aaaaggagaa tatgtccaga tgaggacaaa aagtgtctat36301gaagccagac acctgatgcg ccaactatat ctcatcagtc tggaagcctt acctctgaaa36361cttctattac ttttgctgag caaaaatata tggtgaRtcc taactttgtc taatcattta36421ttcaaaaaga gagttgtttg aacatttaac tatagaaaat atgaaatctg tatctgttaa36481gttaaagcta tatcctctgg atgtgaatct gaagattaaa atttatactt tctaggtata36541agaagattat ctgaaataaa tatatagtga aaaatttaat gttttctttc tgggtttccc36601atttattctc catcatagac tgggttcttt atatcagtag aaataggcat acaatatgca36661tacaaagaga aatgtcctaa taaagtaaat ggaaggtgca tgaaagagaa attctggaag36721acaaatcaat aatatYtagc ttggatatta gagtcacatg gtcagaactg attccagact36781tccaacttac tagtaacttt gtgttttctc atccataatt tgtgaataat atctacctag36841cagcactgta tgtatgagat taaataatga gatatatgta aagcatgtaa actagggtct36901gacacatagc aaatgctcaa ttttcttatt cttaatctta ctgttcttgt catttattat36961tttacgaatt gctatttgta gaacttaatg ttttacattc tgatctatga ttcttatggt37021cggaataatc cacaatagta taaatcttgg tctatgttga gaatttcttc atacaaacat37081aattatgtag ctcttataca ggccaaacct gcattcattt ctattagtta tatgctagag37141gtaatgaatt tttattattt atttatgtat gtagacacat tctattacca ctctttacca37201actgccatta ttttggctta ggagaaaatg atcttgaaaa caggtctatg tagttttaac37261agctcttttg tgaacttcgg atgtaactga atcattttct agatttatct gttaaataaa37321tggtccatga aagtatgtat gggtagataa ttgatggaag caaataatcc tattgaaaca37381tttgctattt atttttgata cagttctttt aaacagactg attagttccc tttatgattt37441caccctcgaa agcacacacg attttgttct ttatcattgt tgcatgacat tccatttggc37501agttgggctt atttttagcc agactatcaa agtatttttc aactggactg tcactgcact37561tgaacttgga atcttataac ttgaagaact gYcctggaga aaggaagaaa cttataataa37621atgggaaatt ataaatctag accaacccaa acttgtactg gtaattattc taattattct37681tatttcctta agtgtaatat ggttcaatta tagttactga taactaacaa ctcattggaa37741tatgtgttaa tttgtatacg gaagataagg cagcatgttt tgtgaaagtc tttgaaaaac37801tatctgaact caaattattt ttttctgaag atctttatag gaaacaagaa atttcttaca37861aaaatattat tttagttcct tggtataatg ttacagaaaa aaatttggaa tattgctgaa37921tgtagggatt ttagaattat atcttaagta tctagtattt aaaaactaaa aaccgtcttg37981caactagaga ttgctataat taaaatgcaa taaaatcaaa tgaaataagt caaatatatt38041tttctatgta aatatattat ccaatatcga atgggttctt cattactaca attcaatatg38101attgcatttt aaatataaac atctgcattt taaacWcaca atcagtaatt taaattaagt38161attgtagaat ctagaataaa aaacagtttt gattacttgt atgttatgat ctgtgtctac38221attttgcaaa acacaactat gaatcaatct catttgttct tctttactag atgaatggaa38281gaaaaaagtc agtgaatcat atgttatcac aatagaaaga ttagaagatg acctgcagat38341caaggaaaaa gaactgacag aactaaggaa tatatttggg taaagttgta agagtcatta38401tttctttgta taagtgtctt cagtattcac cttaWttttt aaagtatctg tatatttttt38461aaacccgtag aacctcagac ataagcagtc atgttctaag ccttaggatg tcagattatc38521tctctttgac cactcatata gctctcaaaa tgcaaacata acatctcacc tccttaaaat38581ataaattgta atctgccatc attaaagttt aaaaataaaa acctaaaggc tagcctttga38641atgcaagcta ttgttgatct cagtaatctt ctcaggaatt agatcattct tgtaaaccat38701gccggcaatg aaaagcaaga tttttcttcc cacatagtgg tcgcatattg gccacattgt38761tctctttgaa ctatgacggt aagtccaatt ttaacaaacc ctctggtctc ttgacattga38821aaggctgtcc cttaatggaa ctgagtatca ttctgcccct gaacaatcat ataacaacat38881ataatattgc cttcaggatt tcagtagatt ttgttccatg ataacaagag aaaaataatc38941taagtgtcac aaccttggaa gacacaaaat gttcagaaaa gaggagcaag acccttatat39001acacaaaaag ccttctggtt aaattgaatc tgacattgag aaactagatg cccattagag39061ggtcaagaaa gttacctgcc tgactaagga aaagttccat ataggccaat tagccatgtt39121catttatagc tgtgttatgt aatgcactca tggcaaagaa ctccctctat tttacttaga39181aaacaagtgt aaagcaatga ccaaatgaca catctttaaa aYaaaatttt agatatagta39241aacatataaa tatcacataa ttggcaaatR aaatcaccca tgtataatat agtttatatg39301aaattcgtca atggatgtaa caaggctttc acaagtgttc aacttatggt tcaaaaagta39361gttggttctc tttatagttt ggcccaggat gatgaactca acactgacac ttgtgatgag39421ataccactga tctgactttt gcaagtagta tcacatgttg agatatggaa ctacatttga39481aattcctgta gaaatatgac tggcatgtta gtacagtaga attataaata tgtatttgtt39541ttgttggaaa agtatgaata tagaaattaa tcataaacat ctagctctta ttgacttaaa39601taaatatgct aaatgaaata tgattacatt gaagagattt tcctcaactt ttaaaggtca39661aagcctcagt tatttagtat tattttttaa tttagaaagt attgagcaca taggtgaagg39721ctagcttaaa ctagatgtat ccaaacctga atattaggat taacaatgga tttaaaatgt39781aggttctttc tgataccaca tcgacaacta caagctgttt tgacaactgc tataatggac39841ttttcctgtg tttctactaa atgcaacaaa tgtgaccttc ttttttctga ttagcattgg39901gggtgagatg caggaatctg tgtcagggag cttacagtcc agaatttgtt gagtgccagc39961tcgtattcaa gtgttgttct ggatacatca aatacaagac attatttaat ctatagtcat40021atgtagattg ttgtcatttc atgtgtgccg aataatatta actccggtgg atgataaact40081aaggattaga gacaaggcag tgggagttgt caagtcacat ggtcagtaaa ttctggaatg40141tgatttgtgt caaggtctgc ctgactccat agatagcgct cttcctacca taccctgctt40201acaaaaacaa gatttaggtc ctgatttatt cattgatttt tgSaatcatt gtagataaaa40261agcatagtta agtaaaggtg tgtaggcaag aatgaatgag taccatagaa agtttaggaa40321aagtgagaaa aaataatgtc ttcaccatat tatctgactt gagtaagaaa gctatgtgaa40381tcaatgtaag aattttggag tcatacaaac atggttttaa atacagcttc ttctatttat40441aagctgtata tccatggata tgttttttaa tctccttaat cctcattttc atctataaaa40501tgaggtgaat aataatcacc tcacaggata ttttcaaaat gaggtaaatt aaatactaag40561tacagggccc aacccatagt aagcattgag tagttggaaa tttctattat catgattgat40621tggataaata aggacagttt gaaggataga agacttatca taaagtgaga gtaacaaata40681gcttattaaa agttagaaag agtgaaagtt tttatgaaga ggatgatttc aaaattaaaa40741atgtaaagta ttagcacaat ttaggaagat gccgaaatct gaaatatagc ttttaatgat40801tttgattgaa agggatttga agaattttga gactagagta ttagaaggct aatcaaaatg40861gctatcaaat ttactgagga taatggtgag acatttttgg agagaaagac caatagcctg40921atgctgatga cattaaagaa gctacagaat gtgatgataa ggaagctgag gagaggatta40981tccagtcYag aatgatgttt ctgcaagaac taggggcctc ttatgttaat ggagatggag41041actgtagtac ttgctaaaat tttcgattcc atgactccac cctttatcat gtgaatcaga41101atctctggag atgtcttcca ataacctgaa tgtttaacaa gctccccaga taaaactatg41161cataataaaa atgatacttc taaagtttga aaactatact tctagaaaga tatccaaatt41221gcactgagaa ctctttgcct gaggtgatat tagtgaaggg atgaggaata tgggaacaga41281aagtaagcaa ctggtgtggt ctaaagagag tcaagagaat caaaaacaag caagcaaaaa41341cagaataatc tgcttttgtc tgatttcgtc aaggtagaga gcataatctg ggagaaaaaa41401gtgaattgct tccagggcag agttctaagg attttctttt ctttggaata aaatgatgga41461tgaaagaaag aaaagaatag tgtgctgaga atgtaaattg tcttactcac tcattcacaa41521aattttaaag gtagttggga agatcaaacc taacactgta tgaaatgaca tttaaatgag41581tatctagagc attataggtg ctaaataaat gttagaatct cattactcac ccttattcat41641ttgtcttata aagagcattc ctgtgataaa caattatgtt atgttgcctg acatcagcct41701ttctctaaac atttagatag caaggagact gtatacaatg cccaaaaagg aacatttaaa41761ctagcatgaa acgtaaagga tcactgatgt aaacagtttt tcatttgcta accaatttct41821ttatggcatt cccggtagat gctaatttca ttcattataa gagatttttc accacataat41881acaccattgc tctatcagct cgaatacaca atcttcctgg catagctttt acccacaaca41941taataaacag caatatagtg ctaaatgtaa gacagaaaga gctaaatagt tatcctgctt42001atgccaattt atagcagctt tggaatggga gcttgatgtg ggagagaggg gaaatggaaa42061tggaatgttc tttcatcttc tgctataaag ttgcattcac tcataacttt ggacatttgt42121ttaggtaatg gattggtgac aggaagaaat gaagaaaaag agaaggcagg aggagaaaga42181aagacagaaa gggcacctta gagtaccatt gtctgattat atcctcctct gtgctgcctc42241taatctttat caacataaca gcacttacta catggtgttg tttttatcca tttataggtc42301tatctcccca ctagtatatg tgttcattaa aggcagcttt ttttcttatt tagccttatt42361tatatttgta cttatttgta aaacaaacta ataacttagg gcttaccata tgccagaaat42421gcttctacgt atctttttgt gcatcagctt atataatcct aatggtgtat atgtaaagtg42481ctttagcaca gtgtctggca catggcaaac cctcaaaaat gttagttaac acttaatcat42541tattgtttat gttaacaacc ctctgaagta agtgttgcaa ttgccatgtt atacctggag42601aaaatgaggc taatcaaatt aaactggctt gaactagatt aaacaatttg tatgatttac42661ctaggattgc atccatcttt atttgatttc aaaatgtatc atattcataa tcttggttat42721gaatattata caaccttttt gaaaaaacta tggaagtgat ggctctccac atgtgacttc42781aaatgttttc aattaaatta ttttgtttca atagttttta tcataataaa ttttaagact42841atgttttgct aatttcattg tcaagccaaa tttgtatctg cttaatctat aatatgaaat42901ttaaaaccat gagaatactc cagaaaagtt ataatttttc tttttctttt tctttttttt42961tttattatac tttaagtttt agggtacatg tgcacaacat gcaggttagt tacatatgta43021tacatgtgcc atgttggtgt gctgcaccca ttaactcgtc atttaacatt aggtatatct43081cctaatgcta tccctccccc tccccacacc ccacaacagg cccccgtgtg tgatgttccc43141cttcctgtgt ccatgtactc aattcccacc tgtgagtgag gacatgcggt gtttggtttt43201ttgtccttgc aatagtttgc tgagaatgat ggtttccagc ttcaaccatg tccctacaaa43261ggacatgaag tcatcatttt ttatggctgc atagtattcc atggtgtata tgtgccacat43321tttcttaatc cagtctatca ttgttggaca tttgggttgg tgcttagttc caagtctttg43381ctattgtgaa tagtgctgca ataaacatac atgtgcatgt gactttatag cagcatgttt43441tataatcctt tgggtatata cccagtaatg ggatggctga gtcaaatggt atttctagtt43501ctagatccct gaggaatcat cacactgact tccacaatgg ttgaactagt ttacagtccc43561accaatggtg taaaagtgtt cctatttctt cacatcctct ccagcacctg ttgtttcctg43621actttttaat gactgccatt ctaactggtg tgagatggta tctcattgtg gttttgattt43681gcatttctct gatggccagt gatgatgagc attttttcat gtgtcttttg gctgcataaa43741tgtcttcttt tgagaagtat ctgttcatat cctttgccca cttgttgatg gggttgtttt43801tttcttgtaa atttgtttga gttcattgta gattctagat attagccctt tgtcagatga43861gtagattgca aaaattttct cccattctgt aggttgcctg ttcactctga tggtagtttg43921ttttgctgtg cagaagctct ttagtttaat tagatcccat ttgtcaattt tggcttttgt43981tgccattgct tttggtgttt tagacatgaa gttcttttcc tttttttttt ttttttcaga44041cggagtctcg ctctgtcgcc caggYtggag tgcagtggYg cgatctaggc tcactgcaag44101ctccgcctcc tggattcacg ccattctcct gcctcagcct cccaagtagc tgggactaca44161ggcacccacc accacacctg gctaattttt tgtattttta gtagagacgg ggtttcactg44221tgttagccag gatggtctcg atctcctgac ctcgtgatct gcccaccttg gcctcccaaa44281gtgctgggat tacaagcatg agccaccgcg cctgacccag aaaagttgta atttttaaac44341actatgacat caacataatc caaaacaact tttttggcat tgagtagata catcttggtc44401agtaattaca gtgcctcact ttaaaattga ctaggtttaa ttatgtacaa aaaagtggtc44461aaaagaatgc tctgttccat taaatagtca caatgtatag aggaaataaa aaaatgtttt44521ctgagataca cctttttgta ccttttctct ctgaaaagtt catctgaggc ctgttgcatt44581ccactatact catccttaaa acaaaatatc ttctaaaatt attgcactat agccaactat44641gaaatttggc tattataaga tgaatagcac tggaaacttt tcaagctgat cttatttaag44701aggagtagac cctttctgtc aaactactaa gtaggatatt gaaggggaaa tgtcatctta44761taattaacat agaatgaaaa tattatttgg catgtttggt agtgtgtttt gccaattagc44821cacaaaacaa aaaataagaa tcgtcaacaa caaaaaaaac cctcaaatcc aaaaggaaac44881aaagaattca taatgcccta agtcagtcta tttctctaat tcttttcttg tttgtccagg44941taaaactagt taagaaccat aattttacag gaatatcttg tgaaatcatt gagaagcaac45001ttgacctcaa tctctctctg cctctctctc tctatctctc cctattgttc tccctctcct45061tctccatctg tagctcgcta tctttctatc attttctctc ccccttaccc ttcatctctt45121cttcttttcc atcctgtacc tccatgattc cttctttcct atccctgttt tgaagagcaa45181aacaatacaa aaacagtaaa aataatatca atgccttaat agaaataact attatagcag45241ctaatattta taaaagcttt acaagctcta acttatttac acttcacaac aatcttataa45301gataacaata attatgccaa ttttatcata tctgtcactc taactataat tgtttattgt45361atgaatggct gaataaatga atatacttca cagtattagt ttgcagattt tatttgcaag45421atatcaaaca aatatatttc tggatatttc ttggagatat atattagaaa cagatcttat45481tgatatagtc tgcttctatt gtttggaggg catggggcag ggaagaatct gcaatgaata45541ggcaagaagt tgcaactagc catttgaaac ttaaatgatc aatttgggaa actagacaac45601ccataaaaaa tccttacctt ctttctacat tcttggcttt tccctaagtt ttaaccaatg45661aattagcttt aaaatatatc tacctgtctg aataccttgg agagcagtgt gttacttttt45721ggccaactgt tcaaatgtta cattttaata agttcccact tgtggtcatg ttagatatct45781tcttgaaagg atgaggtcat gaaaaagaag tttaattgga tcacaaaact gcacactttt45841ctaggtMaaa ggtaaaggct aggaaaagaa ttcattatgc tacaagaaag aaagtaacct45901ttgggtacag aactatcatt tttaatgatt aagaacagca tttactatta gaaatatatt45961cataatagtg gctttatttt aattaggaac ttcattcatg ccccagcatg cactatgaac46021tctttaggat atgttaagtt acagtgtggc atggcaggtg atgaaaaatg taaggggaaa46081ttaaaaaata aattgagctt agggtattag aatacataag tgatgctcaa ttggcagtga46141atttgacata cacagttact ttagaagagg caaaagaaga gaaaatggaa tttctStcct46201aggaatgatt gcttaaatat cttgaatatt ttaattacat gtttcaaaac agaaacgttt46261gactagaagt taatgggaag tgagaattag gattcaaata tagcccccaa agtaactctc46321attttaaaaa tgtacattcc agttattaaa gttgagctaa aaattgtact attaaaaatc46381acacttgact gccaaatatt ataggagtct attctctgtt gattcatttt gttgacatta46441agagaagctg catgagttca aggaggatat aattcaagag gagaacaaaa gcaacaagca46501gcggctcagt ttaaagcaag gaactcttta ggactctata ctgagtcact tatagatggt46561aaagtctagt tcaagattca gttagctgag gaacaattgt tgaataaaga gtcattaatt46621ttgctttcaa ttttatttat gtaaatgata aacattcatc tagtgccttt cttttttaaa46681aataaagcaa tggaccctgg catcttgtat tttctgttta ttccatgttc atttgttttg46741caaagctgat cagagtatta gatatcagga cgaacacagc agtcaatttc aggattttat46801ctagtatgtg tatttccagg actgatgcat tcaaaggatc cgggatttag tgagtagttt46861atatcaagga gactcttgaa aacaacctga agtgagcctt aagagcagaa gtgaggcgta46921tagtaggtgt ttagYaggtg ttgtgccaga cttattgaat ggatcatcaa atttagatca46981cagaataaga aagcaattga gtttaccctc tcaaataaag tgattttcat ctcaacttta47041actttttacc ctcctttgca tttttcattc tttgagtctc tccactgttg aaaatgtagt47101aatattaaaa gttYgagaga tcaagtttaa ataagagaat ctcatgtgca gttctgtgtt47161ctatgtataa tttaaacttt tcagagcacc atgatgaata aaaattatgt atacatagaa47221aacagacatc tgaaagtaaa ctcatttcac tcacacacaa aatctcacat aatatctacg47281gagaaattca ggaaaatcct tccaaaaaga aaggtggggg gaagtagcat ctgataagat47341attggaaaaa agaatataat cccaatgtat tacaacttct gcctgaagca aagttctctt47401gtactctgac tgggtcagat ttcacagata aaggtctaaa ctctggtagg gtactctgat47461atagtccctg gtgagaagtg gagtgattct attaatgaaa tccttgtctt tgtctgaaca47521aagaccattg ccccaggagg gcccttgaga gccacaggta gcctttcaaa aggaatgtaa47581agctaaaatt ctcgacctct tgaggtccac ggtgctccca ggacattttt cagtagaaat47641gagatcatat ccttggcttc acaggtcaat ttcaagctgc ctcctgccta tttaaatctc47701tagcagcttt agagagatag catagtattt ggtgagtgag ccaaagtttc atatcacttg47761taaagtactt acacttgaaa ggtactataa attcagaaca ttatcatcaa ggcatccagt47821taagaacatc aaaagctatt ggaattttaa aatagaaaaa gaaaaagtgc tattctccag47881ggtactgcta aaaagagaaa ggctctaaat ccagggtcat tgaagaacct cagtgaaatg47941ctttaattag aaatggcaag ccttgtggtt tattttcaca gccatgggaa tgcagtattc48001ccaaactttt tttaccctcc tacccttctt gctttctgtg ttcccctgca gcaaaacaca48061tagacatgct tactaatact gggaaacaca cctcaacttg ctttggaaag ccttttgtgg48121aagtacaata aactccagtg tttataaatg ttaataatat aagaaggggg ttcagaattt48181catgaagata gttaagaatt taactcactt ctatcagcaa tttccctcag ctcaaaaaat48241actgcaatca aaatcatttc tgatgaaaat gccactgttt taaatgtcta cttataataa48301aatgagccta cttataaatg atagagctat ttatggtatg gcagtaactt cacaaaataa48361catgattctt agaaatgtgt ggtctttggc ccagcctggc ttttaaaccc tcaaagccaa48421atatattatg gatgtcttgg aagaaagtaa actaggtact tggaacacac ttggaaatgc48481aatcacaggt cacagaggtc ttctttgtgg ttatgacttc aaatagtaga ggtattcagc48541ggagctgaac tctgcctttc aagctaggta gattggattt ttctcttaaa gtttgtaggg48601aatcagtgtc cataaataaa aagggaatga aaactgtgac tggaaactga gaatgttgat48661ggacaRtgat tcagctctga gctgcttcgg cttttcttcc ctggtctttc tattgtccag48721caaatgaaac acactataaa accacaagaa aaaaaaacaa agtattggac tgggggagga48781gagagaggcc ttgtgatcct aagaaaaact atgtaaccag cagcctaagc aaaacaggga48841aggatgcaca ttcttctgag ctaatcataa tgaacttaat ggcaaatact gaaagaagaa48901aaagctttgg agctcttctg tagcaataaa cacctgagaa aatgcagttt ttcaatatgt48961atgtccaata ttgtatagca atacctgtta tgttttgctg ttttcaaagt ggaaaaaatc49021atttcatcag tgtggtaatc cttgaaagcc tcattcttat agttatacca ggattcaaat49081cctatcatga ctctgccttt taagtagttt gttctttttt tccaacttgg gcaagttatt49141taacctattt aagcttcaat ttcttcaatt ataaaatgag taaaataata tatgtgacaa49201tatgcatgac tttgtcctgt tgtgtccgga attggtgggt tcttggtctc actgacttca49261agaatgaagc cgcggaccct cgcagtgagt gttcagttct taaagatggt gtgtccgggg49321tttgttcctt ctgatgtttg gacgtgttcc gagtttcttc cttctggtgg gtttgtggtc49381tgctggcttc aggagtgaag ccgccgacct tcgcagtgag tgttacagct cttaccRcgg49441cgcatctgga gttgtttgtt cctcccgtcc ggagttgttc attcctccca gtgggtttgt49501gatcttgctg gcttcaggag tgaagctgca ggccttcatg gtgagtgtta cagctYataa49561aggcagtgcg gacccaaaga gtgagaagca gcaagattta ttgcaaaaag cgaaagaaca49621aagctcccac ggtatgaaag gggacacaag caagttgccg atggtggctg gggcagcctg49681cttttattcc cttatttggc cccacccaca tcctgctgat tggtccgttt tacagagagc49741tgattggtcc gttttgacag ggtactgatt ggtgcattta caatccctga gctagacaca49801gagtgctgat tggtgcattt acaatcctct agctagacat aaaagttctc caagtcccca49861ccagattagc tagatacaga gtgctgattg gtgcatccac aaaccccaag acagacacag49921agtgctgatt ggtgcattta caatcctcca gctagacata aaagttctcc aagtccccac49981ccaactcagg agcccagctg actttgccta gtagatccca cgcaggggcc gcaggYggag50041ccgcctgcca gtcccacgct gcgtgcctgc actcctcagc ccttgggtgg tggatgggac50101tgggcaccgt ggagcagggg gtggtgcccg tcggggtggc tcaggttgcc tgggagccca50161cgggggtagg ggggtggagg ggccttgggc atggcgggct gcaggtcctg agccctgccc50221catggggagg cagctgaggc ctggcaagaa ttcaagtgca gcgcaggtgg gctggcagtg50281ctgggggacc cggtgcaccc tctgcagctt ctggcccagg tgctaagcct ctcactgccc50341agggctggtg gcaccagcct gcggcgcaga gtgtaggacc cactaagccc acgcccaccc50401agaactcacg ctggcctgcg agcactgtgc acagccccag ttcctgccca ctccactccc50461tccacacctc cccacaagca gagggagccg gctctggcct cggccagtcc agagaggggc50521tcccacagtg cagtgtcagg ctgaagggct cctcaagtgc agccagagtt ggcaccgtgg50581cctgaggagg tgccgagagc gagcgagggc tgctagcacg ttgScacctc tcactgtggt50641gaggaataag taagattaag cgcttactac agtatctggc atgtgatatg caatcaaaaa50701atgattattg cactgttgtt accattgcca acttcactgt gaatatacaa ctttcttttt50761gaagtataac tcttgggata attttagcat catttttgtt gaggcttaga aaattacacc50821tcaaaatgaa ggccttacaa gcagccttag aagccaaact ttctctctga ccttctcctg50881tcctcttgtc tctggcccct cattttcccc agagggtagc catagagact agaatccctc50941tcccctcaag gcaggtcata gaaactagaa cttacagagt tattcctgta tctttgagtc51001ttcattctga aggctcttat gtcttgtatc tcttgtgcct tttaaaacta tgatcaaata51061tattttatgc cttttcttct attaatgtcc cttttgtcag ttgatttcag caaaccttca51121gagaggcaaa ggggaaggtc tttcttggcc cctataattt catgccatat ttgacttttc51181ttggcctttt atatttgaca acaattttct gatttctgat ttcctgattt acataataat51241caattttata tatgtattat gtaagtattc cctactaaca gtgactctca ggttcctaga51301tgatgtctga ccttggtttt acttgtcttt aaaaagtgcc actaaattcg ctattgtgac51361atctagtagc tgtcagtgat tctggagcaa ctattggaat catagtgtta tttctggtga51421catcattaca atgtaaaact acagaatttc ctctcaattt taattcttca tatagagggt51481tcaagattct ttctgtaagt gcaaaataat cttagaaaaa atacatttct gaattgatag51541taacaggatt tgcatttata attttcaaat gaaagacaat atgctaaaag atgaattttg51601tgatcatctg taacatgttt ttttcagctc tgatgaagcc ttcagtaaag tcaatttaaa51661ttaccgcact gaaaatgggc tgtctctact tcatttatgt tgcatttgtg gaggtgagta51721cttgaaactt agtacttctt aaactggtta tatgtgtata tcgtcagtga cttgagctgc51781ttaatagtct gcaaaagaat tctattcata ctcaattttc ccttctgctt tgccttttcc51841aaccttgata atctcagttt aggcagtaac tatcaggcag aatattccaa ttgatttagc51901tttaaatata agcttcattg tgttgctacc ggaaaccaag tccctcagtt tgcttggaaa51961tcaaaaagca cttttcaata ttgggcatgc cagtcaggga ttaacttaaa ttcagaagca52021cttaaagatt ccccaaaata tcatcaagca gttgcttcac aRtattcaac aagagtatgt52081ggaaaagtga taaatgccaa agataagatc taaaagtgaa gattgatcga agtgaagatt52141aagagtttaa tttggcctac actttagttt gttttgttta tcaatgtatc tcaagtaatt52201tgaatggtgc ctggcctgta atagccactc agtaaagtta ttgaattaat gaatgaacca52261ttaatattta ttttgttaat ccaaactaaa ttcattgagc ttaacttctt aaaatggaac52321agaaatgtgt aagtgtcaac tctaaccata ttccttaacc atttctccca aacaaaccaa52381acagactctg cttaacagat tgttctctca acttcaaacg tgccttccca accttggtta52441tatccttcat atttctttga tctaaataag ctctactgtt aataatgttc agataaatcc52501tctacaatta gattttttaa tttaagcttt ttcactaata gtttattatg gtgaattctc52561ttatgtgttt atgtttcaaa accatctgat gatgagggta ctaaataaaa aatctaagac52621agagtcaaaa tacacagcat ttgaataaag ttttataaat ggaaaagaaa tatcccctag52681taagtaaggt tgtcaattaa tagaaaactt tacaaatggc ctgcagaacg cagcttgaca52741tttttataca gctgtatggc atttatcatt aggtcaaaaa gagtctcaaa ctcaccccat52801ccccacaatg atttagcctt ttttcatttt tctctttaag gcaagaaatc acatattcga52861actcttatgt tgaaagggct ccgcccatct cgactgacaa gaaatggatt tacagccttg52921catttagcag tttacaaggt aggacacttt aattcccata aacactgcat tggaactaag52981gatagtgtgt atctttaggc tttttttttt ttttttcaaa ttagtaatca tggaaaattt53041tcagtgacca gaggcgttac attactaaag gacttctttc tctttattga gcagatacct53101gctgcatact agacacatac tctttttaat gctttctgta tttaactcat ttaattctca53161caagaacccc atgagtgaat atgattatta tccccaaggt tacatggtta gccaggggtt53221agccaggatt taagacaagg cagtctagtt ctgttgccaa agctcctcat cagaacacca53281tgatgcctct gtgcagagag agatgaggtt gtatttaaag acaaaagaag cacactgaag53341ctgtatttct ttactgcaaa tttttcttaa aaaacctttt tgctttagct gtttattttc53401ttatattagc ccctggctcc aacctaatct cagcacctga tagaataata aatataatag53461tggattggac aaactgggat acttagtgtt ggaattgaac cttgagattt tcataaatat53521aaatttataa aacatacaaa ttatattatg cattgatcat tttataatta taataacaat53581ttctactttg tatttattat gagctaaata tcatactaag tttaatgtat tttagtcatt53641aaaataactg catattagga tttactttac agaaaatgca attaaagact aaagtagata53701cctggacaat attagaatag tgattaaagt caagtacagt tggctcatta ctcccagaat53761cttatttgta tattgcgtat tttgttcttt ctcattttca agcgagtctg tgacagaatt53821ttatttcagt tttgtgtgtg cacgtgtggg aggagggaca actttttgct tgtattagga53881accccctgca ctatctccct cccccaactg ctagagagtt tgtcattggg agggaagagg53941accttatttt cccatgatct agcatcacaa attcctgtaa ttcatagcgc aaatactcta54001aagtttcatc tcttttcatg cccagtcaat gcctagtcaa ttcttctaat tccttctggg54061agaaagcctc agttaagtgc taaattgtct ttagtacctg ttaattatcc tgctaaacta54121tgaaacattc tgagtttcca gtgtagaaaa cagcatctga tgttgacttt tcttctgaaa54181ggcaggaggt gaaagtagaa gagctccaag gaacaggaag gtagtgaatg atagtcatgg54241cactgtatga aaccatgaat tctgaacctc accgtctggg agattttttc tgtattagtt54301cattttgcaa agatttagca tgacattggc acacaaatgg gttctgaatc atttagtcct54361acaatattta gagaatatat ttcttcaacc tctgtttata ataattaaat ttccactttt54421agagttttct ttgctctatt tttatgtttt tttctatttt agagaagggt gcataaacta54481taactttttc atagcaagtt taaatgaaaa tttagattct tgggaaacaa agcaggcaaa54541aatattaaaa ggccataaga aaatgcatag ttattttgca aatttcttaa caaaagtgaa54601aaacaaatta aagttactat ttcattttaa acacaaaaat aatataaagt agttgtgtgt54661ccagaattgg tggattcttg gtctcactca cttcaagaat gaagccaggg accctcgcgg54721tgagtattac agttcttaaa ggcggcatgt ccagagtttg ttccttctga tgtttggatg54781tgtttggagt ttcttctttc tggtgggttc gtggtctcgc tggctatgga gtgaagctgc54841agaccctcgc agtgagtgtt acagctctta aggcggcacg tctggagttg ttcattcctc54901cccgtgggtt cgtggtctcg ctggcttcag gagtgaagct gcagaccttc gcggtgagtg54961ttacagctca taaaggcagt gtggacccaa agagtgagca gtagcaagat ttattgcaaa55021gagctgaaca acaaagcttc caaagtgtgg aggggcaccc gagtgggttg ccactgcttg55081ttcaggcagc ctgcttttat tctcttatct ggcccccacc cacatcctgc tgactggtcc55141attttacaga gagctgattg gtccgtttta cagagagctg attggtccat tttgacaRgg55201tgctgattgg tgcatttaca atccctgagc tagacacaaa agttctccac atccccacta55261gattagctag atacagagtg ccaattggtg catccacaaa ccctgagcta gacacagggt55321gctgatggtg tgtttacaaa cgttgagcta gatacagagt gctgattggc atatttacaa55381tccctcagct agacacaaag tttctccaag tccccactag actcaggagc ccagctggct55441tcacccagtg gatcccgcac aggggccgca ggtggagctg cctgccagtc ccacgccatg55501tgcctgcact cctcagccct tgggcagtca atgggatggg gcactgtgga gcagggggca55561gcacgctttg ggtaagctcg gccatgcaag agcccatggc agggggtcgg ggggaagctc55621aggcatggcg ggctgcaggt cctgagccct gccgcacggg gaggcagcta tggcctgctg55681agaaatggag cacagcgcca gtgggccggc actgctgggg gacccggagc accctccgca55741gctgctggcc cgggtgctaa tcccctcact gcccggagcc ggcaggactg gccggccgct55801cggagtgcgg gcccgccaag cccacgccca cgcccaccca gaactctagc tgggccgcaa55861gccccgccgt gcggccccag ttcctgcccg tgcctctccc tccacacctc cccgcaggcc55921gagggagcca gctccagcct cggccatccc aggaaggggc tcccacagtg cagcagcggg55981ctgaagggct cctcaagtgc agccagagtg ggcgccaagg ccgagaaggc accaagagcg56041agcgagggct gcaagagctg ccagcacgct gtcacctctt agttgttcac actaccaggc56101attaaaccta gtaaccatta aatgcattaa acctacttct aattatccct tgacagaact56161actccagtaa gtatataatt cattttgggc tccagttccc ttcccccata tagtattaat56221aactgtttaa ttttaatatt atttcaaact tacaaagaca attgcaagaa tagtacaaga56281aactgcttat ccaggttcat taactgtttg cattttgtcc catttccatt attttttttW56341agaaccattt gacagtaaat tgtataaaac atgccctgtt gccacttcag tcttcatttc56401ctaagaacaa gaaaattcac ttacataact ccaatgcaat tataaaatta aaaagaacat56461tgagatgata ttaacattga taagatatga taatctatag ttcaaattca agttttatca56521attgtcccaa ttgtgtcatt tatagattat attttcttgt ccaggatagg attcaatcca56581caatcatgca ttttcagttt tcttatttct gtaatctccc ttactctgaa agaatttctc56641acccgatcct tgtatttttt tatcttaata tttttcatgc ataccagtta tttatacata56701tttctttgaa tttttttggt attgcaatcg aattttattt tgttcataac actcacaaaa56761tgaataatgt aactctccca tccttttgtt cactacatca aatagctctt tcaattatac56821accctggcat gctgggcctg actcattgtt agaggaaagg ggtcccgatc caggcctcag56881gagagggttc ttggatctcg cgcaagaaag aattcagggc gagtccacag tgcaaagtaa56941aagcaagttt attatgaaag taaagtggtg aaaggacagc tactccatag acagggtagg57001atgttcctga aagtaagagg aggagcacat ccaccctagg tacaatgctc atatacatgg57061ggagatgtgt tctgctgcaa gggtttgtga taacatatta attttcttaa ttgctatatt57121ttgcaagaat cgatattatt atctttaaag caaaattagg aatgcctttg ttctacagat57181atcaggatat ctggacactc ccaagtctgg gtctgttcag taaacattat gaacttgttc57241tttaactgta aacatctaga ggttaaaaat gcctaacttt tgggaggccg aggcgggcgg57301atcacgaggt caggagatcg agaccatccc ggctaaaatg gtgaaacccc gtctctacta57361aaaatacaaa aaattagccg ggcgtagtgg cgggcgcctg tagtcccagc tacttgggag57421gctgaggcgg gagaatggcg tgaacccggg aggcggagct tgcagtgagc cgagatcccg57481ccactgcact ccagcctggg cgacagagcg agactccgtc tcaaaaaaaa aaaaaaaaaa57541aatgcctaac tttctgagaa tgtaaccccg taagtctcag cctcattttc ctattttcct57601agccctcact caaaatggag ttgttcactc tggttcgaac gcctctgaca tcatgactgt57661atattttcta atcaatagag taatctctgc tttcattaat aaaagtgaag cagatattta57721gctatttaat accacatgaa tgattaatcc tcatacatct ttttgtttta gtttgttgag57781tttttcacag tgtttagttt tattgaggtt taatatatac caaatacaat tatcaatttt57841aagtctacaa ttgttagctt ttgaaaatgt atgtaatcat gtaaccatca tcacagtcat57901gttataggac atttctacta taccaaaaag ttccctcatg ccattttgca gtcagtctcc57961ttctcacctg gtctctggaa acaagtgttt tctgtcacta tagttttctc ttttctagaa58021ttttaaataa atttattaca tccagtatgt tttcttttct atatggtttc ttgcacttat58081cataaggcta ttgggattca cccatgttgt atattctgta attaatgctt ttcttgtaga58141attgtattcc cttatatgga tataccacaa tttaccattt acctgctgat ggacattttc58201ccagattttg accattataa acaaagctgt tctgaacact cacatacagg tatttgtttg58261aacatatgtt tttatttctc ctgggaaata cctaaaagtg gaattgctga gtcacataat58321aagtatatgc ttaattttat gagaaactaa caaacttttt aacaaactag caattccttc58381ctacattccc atcagcaagg tatgagagct gtagtttttt cacatccttt ccggcacttg58441gtgtggtcag tctttttttt tttttttttt tttttttttt tttttggcta ttctattgga58501tgaattttag tatttcatta taattttaat ttgcatttcc ctgaagacta aagaggttga58561ataggttttt atgtgcttat taggcatcct ttatttttta atgtagtgtc tgttaaatct58621ttttgccatt tgtaattttg tttattattt tcttgaattg tgagggaact ttatctattc58681taatacaatt cttttcttga atatactttt gcaaaaattt ctctcagtgt ctgacttgcc58741ttttcatttt cataacagtg tcatttgaag agtaaaaggt ttttgcatta tttttaaact58801tatgactgta gtgattacca tacagttata gattctcaac atttcatagt ccactccctg58861caaatgttga tctacttcac ttaaaatata agaaccttcc agtagtatag tttcatttta58921tcttcattct ttgtgccatt gttgccatat acattgcatc tatacatgat aagaacctaa58981atatattatt agaatttttg tgttaaacat ttatatcttt taaagaaatt aataagagag59041aaaatatgct tttcggaagt ttgattataa taatcctaca tatggttttc tgtatttaaa59101actctttgag attcactgag tttttatatc tgtaaattta tggtttcatc aaattcagaa59161attttgacca ttagtttttc aaatacatat gttagattgt ttgataattt tccacatatc59221actgagtctg ctttttttct tccttcttct ttctttctgt tctccaaatt agattatttc59281tacttatttg aattcaagtt cacaaaccac ttctctgcca tctcccatct gataacaatc59341caccagtgaa ttattcattt tggttgttta cttttccgtt ctaggatttt tatttattaa59401gattaattaa gatataattt caatttctct gctatgatga cacatctgtt atttcatttt59461tccatatttt ccttcaaggt tttgaaccta ttatttgtga ctactttaaa tgtccaatct59521gctgttttca acatctagat tagctcagta tttgtttcta tttcatgttg actacagRgc59581acatttttct gtttcttcat atatcaatta attttttatt gaaagctaga aattatgtat59641tatgtgttat aggaaatctt tatcatgtca tcttccttta aagaatgttg agtttttttt59701atgtttgttt ttctttcatt tttggcgggt agttacactt ctggtggata catttcaatc59761aaatcagtct tgatttgatt ttttgttagg ctgaaggtat ttcagtttga acttagcctc59821ttggcttacc tctttttttt tttttttttt tttttgaggc ggagtctcgc tctgtcaccc59881aggctggagt gcagtggtgc gatcttggct cactgcaagc tccgcctccc ggttcaagcc59941attcttctgc ctcagcctcc ggagtagctg gggctacagg cgcccgccac cacgcccggc60001taattttttg tatttttagt agagacgggg tttcaccatg ttagccagga tggtctcgat60061ctcctgacct tgtgatccgc ccacctcgtc ctcctaaagt gctgggatta caggcgtgag60121ccaccgtgcc cggccgcctc tcggcatacc tcttattcct agcatgtggc tctcacacct60181gaacatggtg cttactccta aagtatgatt ttttggcatg tcacctgaat tcccaagtct60241ctcagtaaag tctcttcata ccagctgaca ggagttccaa aatccccccg tactacatga60301cctccagtat caccaccatt caccttttag caataaatca ggtgatcttt tctagatctt60361ccagaatctt tctctgtgct tgtgcagccc agctcaacaa tacataatgg atctccatgc60421agatttctgc agtgccctct ctgtgcagct tcttcttctc ggttaccctg ctccacaaat60481acagcccctt cataatatca gaactctcat ttacactctt aactcagtga gacttcctat60541tatgctttgt ggcagaaaag tatttcaggc agaaaaccat ggcaatcatg ggatattgtg60601tgacttgctt tatatctttc tcttaaagga cacagctgtt tgctcatgct cttcagtgct60661tgagaacagt tatgtcatat attttgttcc attttgtaga ttttatttgc cagtagagaa60721agtctatata agttgttctg tcatgcccat atctactttt gcattgcttc caaaatgatc60781Wgaatgaaac aaaaatctat gatatcattg ctcactttaa cacttactat tgtcatccat60841tgcctatata atgacattca aactcctaaa tatgagactc aagatccctt aatttataac60901ttttctggaa tcactttcca tactcttttc ctgtagtctc tgcctgttca accctaatgg60961ttttcaatgt aatgtgcatg aggatcatat agtaacatac gtttaaaatg aatatttccc61021tttctacttt actctcattt aaattggtac atctagggta ttctaggaat ctaaattttt61081aacaaacaac ctaggaattt tgatgtaaac aatctaggtc tacatattgt ggattactgt61141tcaagcgtgg caactcaaat tatggtcaca tgatcagtag catccacccc aactaaaaga61201tttcagaaat gctgctctca tttcctttcc agccctattc aaccagtact attcaggcgt61261ttctagagaa atttaaccaa taggattaat ggatctatct atctatctat ctatctaatc61321tagagacata tttattatag gaactggctc acactattat ggagcccaaa agtcccacga61381tcttccgtct ataagctgga gaatcaggaa agctgatagt atgatttggt ccatgtccaa61441agacttgaga accaggggag cctatggtgt aaatcctatt ctgagtctaa aggcccaaga61501acgaggatca gtgatgtctg agggcagaat atggatgtct cagctgaaat aaaaacaaat61561tcaccttccc tctacctttt agttctattc aggccttcca aagattagat gatgcccacc61621catattggtg agggtgatct tctttttcag tctactaatg aaaatgctgg cagggcatag61681tggctcatgc ctgtaatccc atcactttgg gaggctgagg caggtggatc acttgagctc61741cggagtttga aactaacagg gtgaaacctt gtctctacta aaaatacaaa aattagccat61801atgtggtggt gtgcgcctat agtcccagct acttgggagg ctaaggtggg agaatcactt61861gaagtcagga ggtggagatt gcattgagct aagattgtgc cactgtactc cagctgggca61921atagagcaag accctgtctc aaaaacaaac aaacaaacaa acaaaaaact aatgccttct61981ggaaataccc tcacagacac ccagaaataa tgttttacta gctaactagg catcccttaa62041cccagtcaag ttggcaccta aaattaagca tcacaaccag aatgtacatt tttaaaaaaa62101tctcagctga tgtatttgta gattaaagtt tgaagattgc cattttagat aactccattt62161tttaggccaa acagttccac atgtctttat ttttgttttt ataatttctt ctgcttaaat62221attttttaat ttcttttcct gatgcaaaat tattacttat ttaaaaccaa gataaactga62281gttttattga actcgcagac agaaattatt gcactctctt ttgggtttca gttattaagt62341ttacattatc ttttattgta gcacttataa ttatttattt atttgcatat ttcaaatgtt62401ggtctgtgaa ctttggacac tgggactaag tctttttctt tgattcctca gtatatgtga62461cctgtaaagg atccttgaga aatacttgtt tggaaattag tgagtgtcca aacattagaa62521caatacttga catttgttga tattatgtag cttaagaaag actgtcagct tagaagctag62581aatacttctg ttcatatctc agttatgaga cttgctaatt atgcatattt aggcaaattg62641agttttcctt atgctcctca attttctaac ctctaaaatg aagcagatag aattatctat62701tttattgtgt agtttagagt tttaaggaaa caaatcatat gcagtactta gcaaaatacc62761tagcatatag ttaatactta ataaaaatta accattattg ttgttgaact ctaactttga62821caaacattta ttggaaacta acatgttttt cattttgcat agaaaaatta aaagaaatgg62881attggatttt ctggattatt acattctaaa gagatgagaa aaggtaatcc taacaatata62941aaagaaatgg catctatact tagatcaatg gactacaaac atcttttatc acgtcctcta63001taaaattagc actcccaaac acatatatgt gttcataaat tatatacatt aacaaaaaaa63061cccatcagtt aatagcttaa gacaaaatct actctttgga aaacaaccag tgttaaagtt63121tgtagctata catttatatt tcaaatagcc tttttgataa tataaaatat tttgacaagt63181ttattacaaa tatgattaga ttaggaaatt gatttatatg aagctcagat tggtagaaaa63241aagtaccagc tttttctttg ctgcttctta ccttagtaat atatatgttg tgtattttct63301tcacaggact cttttaaaga tacctgccaa tttcgtctat tcagttgaag tgtctattca63361gttaaaattt gatattaaaa tttataatcc cacttacttt ggctcacata agctacagca63421catccccaat aactaaaagc taagaaaaaa aaaagagtga gaggcagctc tcagtccttt63481cccatgtaga attttaagta tatattatac tttgggtact atatgtgaca tatgactagc63541tgatatatga actgaagccg ccagtgtaaa tttacatgtt aaacatgaat gaaacttcat63601ttatctttat atataaatgt atataagcag attttctaat tctttttgta ccctaatgaa63661ttttattaca agattcctgt cacatttata tccctctata gggcatactt ctttaggtgg63721tatctaaatt gtattcctaa aatgaaacac aagactttgg ggaaaaaatt gagacctctg63781atatagttta gaagatctta attgagaaag ctaatgaaca aaatcctatt agaccatcta63841atttagcctt acatgtaact ctcattcaat acacagacaa gccacttagt gaaatttctt63901tagtgagaaa gaagtagcat tttgattcat taacttattt ttaacatttc gtatgataaa63961agcagatatt gaatggtaat ggtcatttat cccatcctat atatttcttg caagatttaa64021ttcctatcat tcagaaacaa ctcattttat atatgtatta caaaagggaa ggacatgaat64081gggggaaaaa tcgaattaaa cctatgttgc ctggcttacc agacctaggt attttttgct64141acgtaactgt gcccatcaga aaactctgac ttttctatca gtggcgattc gtagcctgaa64201tcattgtatt aatatctttt atcctgactt tttttttcct ccttagcaca tatatttgat64261tgcaaagttg aaattttcaa ctatcctgaa actaaagacc caaagtaagt gtttcatagt64321ttctaactgc tgtcgattta gaccaaggtt caacagcctt ggcactactg acaatttgga64381ttggataatt ctttgctgtg ggaggctgtc ctttgtgtta taaaaatgtt tagcagcatc64441cctgtcctct gttctctaga tgagagcagc cacttccctc atgcccagtt gtgagtacca64501ataatatctc cagatatcac ccctgagagg caaaaatcac tcccagttga gaacaactga64561tttagaggac agatttcaat attcattaac ttaaatggat tttttaaatc ttaattacct64621ttatatttga acaaataaag tttcctgatt aaagactgca ttttccctaa atgggaattt64681gacaagatag taataagaaa taatttgaag aaaaaaaaaa ctcaaaaaaa aaagaatccc64741actttttgca tatttaacta aattctccag ttagtgatga cttttaaaca ctttaatagt64801agattttcac ttaagaaact agagaaaatt tttactcata aattacagca atcattaaga64861tagcttgttt atatgcagat ttccctttag taatgaaata tttcataatc tctttctaaa64921tgaacacatt ctcaataaaa agttaatagg atcctagctg aagatggtaa 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attgaggagc67501tccagaggaa aaattgccta tgRaaggagt gccacacggg ctagaaatgt cgaggccctt67561cttttaccac caccttactc agtcattgac tgggagtaag gtccaccctg agaagagact67621gaacacagct ctaatgctgc agtgggtcct gaaataactg agaaagctgc cagtgaacta67681ccatcctgcc acactcttca taaagggagg tctcagtggc atatctattt ctgccacaca67741tgaggaaatg ctcatgattc tataatatat gaaaaaagga aattacctat atgtagaatt67801agatcaatag tgtacataat aacaattatt atgtgtcaag cattgtacta aatgaattac67861agcattatct gatgtaatcc tcacaacaat gctatgagaa tgatacaatt gtatcttcat67921tttacagaca aaaactggac agctcaaaag gttaagaaac atgtcgcaaa tgtaggaagt67981gcccatcaat attgaaacac gagtttgctt tgctgggata gcttgtgctc catctggttg68041cacaaacata tgtacatagg acaaagaatg gaagggaaaa tacacacata aataatagtg68101attatatggt agtactatgg gttattgaat tatttattca ttttctttgt tttttacatt68161ttcttctctg aggtattctg gttgtctgtt gaggcataac aaactatccc aaacttggtc68221ttctaatgca actattttat tattctcatg gattcttagt tcaaaaattc agacaggatg68281ccgtgtggat ggcttgattc tgtcccacaa tatttagagc ctcagctgag aaaatgtgaa68341tgaatgggtt tattcatcag ctggtagctt ggaattctct ggaggcttaa cacactgact68401ggaataagtg taaggctgga cttagctaga acagttgatt ggagagcctt catgtgcgat68461gtaccttctc catgtggcct gcattttctt ataccatggc tgacacaggt tagtcagatt68521tgttacgtta tagatcaggg ctcccagagt gtgtttttca gcaaacagaa aggaagctgt68581gtggtcttct atgtcctagc cttagaaaag aaacactctg ctccacgcta ttgatcacaa68641tccacccagg ttcaagggga aggggcatag atagcaaata tctgtgggtt gagtatagat68701gaatgctcac catgttttaa aaccaccaga aggtatattt tgtccataat cagaagaaag68761tacattttta tacatcagtt tcttttctat actgaaatag agttgaaatt tttttgttta68821ataaataaat tttaaaaatg gatgtaatat ggtagtagag gatttgagct ctgtgatata68881gtggttgctt atcttggctt tatccatagc aataatcaca gagctaacaa taaggcagga68941tattaagtaa gcattgtttt agagttttat ggctagtaca gagttgcagt tactaccatt69001acaaaatatt tttatccatt tacatacatt ttgggagatc tattctacct caaattctat69061ttctcaagga tatagctgga gYgctttttt gtctaagcct cattcttcca cacatggatt69121ttcttctgct taaaaataat aaatatgaca cacacaaata cattacaaag 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ttaaaaaaat70021tccacaaata ttgactgaat aaggagaaaa aacaaagata cttgaagaaa ggataaatgt70081gtgtttacta aaactgccta atagataaat gtttcacatg ctctgtgatt tacataaaat70141ttattttaat ttagcctagt acagtgcttg tcataaaata gacattcatt aataaatatt70201tgttgaataa aagagtatcc atctttctac tgatttatac aagttattta tgtattaaat70261atgtaaacaa tttgtcagtt ttatatgttg tgaatatttc tccaactcgt cacttgcctt70321attgtgtttt taacacttta atttgctcaa atctatcata ttttctattt aaaatataac70381tttatgctta aaaagacctt cectactact ctgcattgaa atttctgacc atctgcMggg70441aaaatgtttt aaggcgatgt ttttcctatt tatctcaaga ctaatcatgc ctttagccct70501caccaagatt attggtaaag tgagagttaa ctcctgtgaa ctgttttctc tggaggagga70561attctcagtt tctggcagcc tatccatggc taaactagta gacgttttct cgcttccagt70621actggtacag cctttttcca tgttttattc ctttgtttgt tgcagtgcct aactcgtgaa70681gcagagggag tggagttagg aggaagaggt aaagggaaag gagaaagaga agctaagtga70741ttaaatcaaa tcaatatttt atttggaagg tactataggt attatcagaa caattttaag70801gtggttacaa ttactctttt tttatataat caaggaaatt agcaaaaagg 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cagtgggaaa72541aaaaaaacaa aacttttttt ttaaaacgga gaaaacataa gtttatagtt caagcatata72601gagccaatga aagaagaaag gttgaagatg caatatttat ttcactttct tgggagagca72661ggtagaaaaa cgggaagatt tggagactat aacattaatt ttactatttg tagctcttta72721ccaaagtttt ttcatttttt agaatttctt gtaggccttt atgcaggtgt gacagcatta72781agaaatcaaa ctttgccttt taatggtaaa attttcttaa caaacagtat ttgatgttat72841atgtatgtaa gtcataaatg cactggccta tcttatgact tgactttatt gctacttggt72901gtgttgatag accaaatatt atagaagttg tggtttaaat tcctactttg aggcttaaac72961atggaaatct ttgtataaca aagattaaat acaaaacaaa attacatatt gaatttcaaa73021ataaaaattc aatgactgtt aattgctatt aatattataa atattatttc atgtatctcc73081aaaggcagaa atacaataca aataatatta taaaaaagta agaattactt taattgttgc73141acaaattttt caaatttgga ggctcaaata atagttctaa catattcact ggtaaaacga73201gtcaagttaa gaaagatatt atgaattaat tgtgcctaga tttaatgata ttatttatta73261gtaagaagaa attactgaac tatcatgaaa cagcctactg ggctgcatgt actttgttca73321gtacttggtt catactctct tcttaaatgg catatttcat cctcatctct aaaagaaaaa73381aaataaacct ttaaactgtt actgtaaatt cctgacctca ctgggagtta attctagttg73441ctggaaaagc atattcattg gagtaaccta cctgtaaaac tgagtttgtg gttgcaagag73501tctcaccata ggtaatgtcc tttctgtctt tcttgttcaa gcggtagagg attgattaga73561actgaRcacc tctgtcattt ggtcctactt tccaccatct gagaactatt ctctggtatc73621taatgtgctc ttatgctaga aatcacactt aagtttcctt tgagcacctg aactggctga73681taggcagaaa atcaatacat cctgttgcac agcttttgaa cctgttatta gcagaaaagt73741aacactaaag atatgtttcc ttacaggata atgcagaatt gatcacttct ctgcttcaca73801gtggagctga tatacagcag gttggatacg gtggcctcac tgccctccat attgctacaa73861tagctggcca cctagaggta agtcatgcct ttggcacctg tgaaaacaaa ggttatttac73921cttttcggtt gttgttctta actgttttga aatactgtct ttgagacttt acaagttggt73981ttatttttta gctactgaac acattggggt aaaatttagc tgtataaatg agctttggat74041agtcaattca gtcaattttc taacagtgta tttaatcatc aRttaacata aaggtagcta74101gatatacaga ggctttatag cagctcagtt tccacaactt gctattttaa ccttggggaa74161gttactgaat ctcatgacct ccctcttttc ttcacttata aaataggaat 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aatcctgatc75061attttactca aaatcccatg attgtcacac tctatcaatt gtggaaacac caatagctca75121acaatgtaaa agatgagaag tctgacatcc ttattattaa atagttatgt tccccctttt75181agtcatgttg caattcaatt ataccttctt cactaataca cacacatata cgcacacaaa75241ctgtgaacaa atagagctgc atataaaatt atacttgaag cctgtaaaac agaggtgatc75301catctccctt aacagtgtca cttcaaaggg ctcttacatt caatacaaat taaaatatct75361cgttgagagc aaaatagttt ccttcatgtt gacatttcca ctcctttcca tatcacctta75421tttcctggaa aataaatcct tttagtgaga ttcattgtaa atataggtga gtttgtgtta75481cttatgagga aaggggaaat tccattagat tcacctaaat ataaatgttt tccaaaaaag75541aaagaaagaa actgaggcca gtgataaagt aaatgagtgg taaacattaa gggatctggg75601tgaaataatt tatgagttag acactaactt ctagtaaaac atgatcaacc tcttaatgcc75661aggaaaatta cagactatgt cttcaaaaca aaatcctttt tttcctaatg ttgttttgat75721gatagcatta caatggcttc taaagcaaag tggttaagag cttagtttct tcatctccta75781ttcaaatagc tctgttttct ttctttctaa atcataatat ggaagaaggt attatattga75841ttttaccaga cttagggcaa acttcaccaa agtattccaa atagatctct actcttgcct75901tgcaattctt catgggtttt ctaattttcc tcattagctg atatttgatg acttttatag75961tcaatagcta acactgcaaa aagcaagtat agatacatta gcctttattc tcttcttata76021tgggggacta attaaattat tgatcaaata ataaagaaaa gggtatccaa aactatctaa76081aataattata ttcaaaatga atatttttgt tggagtattt ctttgaaaac tattaccatt76141gacttttgac caatgtggga gttaggagca ctgatcccct gtacagttga aaatctatat76201atacattttg actaccctaa aacttaatta tttatagcct actgctgatt ggaagcctta76261ttgataatat aaacagtcca ttatcatgta tttttaatgt tacatgtatt atatactata76321ttcttaaagt aagctagtga aaagaaaatg ttataaagaa aatcataagg attagaaaat76381atatttacta tttattaagt ggaagtggat aatcataaag gtcttcatca ccattgcctt76441cacattgaat aggcaaagga ggaggaggaa gaggaggggt aggttttgct gtctcagggg76501aggcagagat gaaagaaaat ccatgtatat gtgaacttac atagttcaaa cctgtgttgt76561tcaagggtca actataattg tagtaaatag tcactgtcaa tatatcttta cccatacgtg76621tgaattcatt taacattttt tcagccccta ctttgtctat aaatggttag aatatagaaa76681tataggaatg atacaacaaa ataagacata gatttttccc ctaaatattc tgtaatgcaa76741ctgggcacac aaaataaata tgcagaaaaa atatataagc aattcaaatg taaaacagca76801atatagttat aagtaaggag taggatggat gagagtaggt ttgataattg ggaatacagg76861tgttcctcaa cttatgatgg ggttatatct gagtaaatct atcatgagtg gcaaatatca76921taatggaaag tgcatttaac acacctagcc tactgaacat catagcttag cctagctgac76981attaaatgtg ctcagaactc ttacactatt ctacagctgg gcaaaatcat ctaacagaat77041acctcttttg taagaaaatg ctgaatatct tatgtaattt attgaatact gtactaaaaa77101tgaaaaagca gaatggtcgt atgggtactc aaagtatagt ttccactcat atgtatcact77161ttcacaccat tgtaaagtca aaaaattgta agctgaacca tcgtaagtta gggattatct77221attattagaa acataataat gtagagaaaa tgtcagagaa cacaaaatag aagagttaag77281agttcagtga ggatagagat tgtattagtc cattttcaca ctgcttataa agacataccc77341gagactgggc aatttacaaa agaacgagat ttaactggac ttacagttcc acatggctgg77401agaaggctca cagtcatggc agaaggcaag gagaagcaag tcacgtctta catggatggc77461agcaggcaaa gaaagaataa atgagacgaa aaagcagaaa cccctgatca aaccaccaga77521tttcctgaga cttattcact gccacgagaa cagtatgggg gaaaccaccc ctatgattca77581attatctccc actttgtccc tcccacaaca catggggatt atgggagtac aattcaagat77641gatatttggg tgggaacaca gagccaaagc atatcagaga ttaatgtgga ctcaaagagt77701tagggaagga gttatgagag gggatttaaa attgaactag tctaaagaat gatgagaact77761agacagcaaa aataagaggg atctggccca aaaataggaa ggaaagaatg agataagcac77821agtaagggac agtgagtaaa atgataacat gaaacaaatt gtgaagagtc cctctaggta77881aaattaggtg ttaagaatag acgagactaa agaaacttgg agagtagaag tctaaggcgt77941ttaatttaat gcaagaagca attactgtgg tttacaatgt ataacacaat ttctgtggtt78001tacaatatat aacacacaag agtagatggc atggcagatt aagagattag tagaacacag78061aatttctcac tctttaaaga aaattagaag aaccaatgtg atttttcccc cataggaaac78121agggactaat tgagaaaata tttcagcaag agaataattg gagatttgat aagtaacatg78181tttaaaagga ttactttact ttgactgcat gatttagcat gaattctagt gcgattacat78241gatgattaaa caaagaaaga taaaagcttg ggtaaaggat tcaatttgga ggaaaaatca78301attagctcca cacatgttga ttttctgatg acatccaaat tttcaagtgg aaatgttcat78361taatcattta gaaacataca aacggcattt aatggacaca tttgggttta acgtaaattt78421ttagaaatta tatatagaga ggtaatattt aaaaccataa taacagctta aagttgtata78481ttgggcataa tggatggtca taggaccgag cctaagagaa tctccccgga gacttaagga78541attattaaat aaggaattat tgaaggaaac aaagaattgc tcagtgatat ggaaaggact78601aagattattt taatattgca attaggagag tttcaagagt taaaggatcc caggtatcaa78661aaaagttagt gcagaaaagt agagcacatt gggataaaac acacatcgag tatttcctta78721ctatgctaaa aatttaggtg cttattccac tcctattttt tattatcatg gttagcagaa78781atagcaacgg atcagtgggt gttctgtttc attactagtc actatctttt attgcccacc78841tttctctgtg aaacaatagc tgatgctaag tctgacagga ctctttgtct ccacatatca78901aattcagacc ttccattagt gtctcaagtg tgtttgtcct cccataattg tgaattctat78961taaacaatct tacaataaaa tttacattga ctaggcagct gtcttcttca ataatagaag79021agaacatgtg ctgcagattc ctagacatgc tgactacaat ttgggactca gatttaaatt79081tactgtgctt tgaagagtag gcaaatatta gttgtcccag gtctcaatac ttctctttga79141ctgatacatc catctatcaa tacatttgca tatgcttctt tgtacaaggc ctaagatcct79201gtcagcaatc aataaacatg cattaaatcc tctttttccc tttaggtttg gggaatgtga79261gtcatacgct ttcccaggca tggaaaagga aaaatattag ctaacatttg gtggtacttt79321atcattcaca tacatttgtc atttggttgt tttgtgagat aactactgtc agactcagtt79381tattagtgag gaaactgagg actggagaag ctgaatgaca tctacatggg cattcagatg79441ataaataaat aagctggatg ggactccaac ctggggatct accttcacat ctgtagtctt79501atacataaaa gagcgattRc tccccagaga ctcttctgtt tattgctata acacagctta79561tacccacaag acacaaaaaa agtatttgga aagtgaaaaa agaaacggaa tatgttacta79621tataaagccc aagcattact acatatacga ttaaagcaac attaaatgac tcaaaactaa79681tttcataatg cattcactat atgggcataa cctatggaag tataatgtag gtatatgagc79741acacaacaca cacacacctc aagtgtaaaa cactttatac ttaaataact ccttaaaaag79801ttgttactct ggtgttacgt gctgtagttt ctgcagttgc cataagtcaa atcagttttc79861tgcagccgtt gacaactgtt tgagcagtga acagaaaggc atgcagcaaa catgtaaacc79921aatgtgtaat gtatagaggt tgtggagtgg ttttcttatt acagaaaagt gtactttcat79981atacaccttg tgaaggtgag cgtattattc atagttgaag ttattctatc ctaccatatc80041ccattttcaa attgtaaaca cttattcaat cactgcttca gaaatgcttc ccagttattt80101tggaacttat ttctgatttc aaggtaatgc actcagtatt gtttcagttc attctttact80161aaatcatcca aatgtttcag tagagaattt tgtttaccaa gaagcctcRg accctgtgta80221agttattttt taataatcgc tttagtttcc tctccaaagg cacaaatgac agaatgctta80281gaattataaa atatggtttc tctctctctc ttatttttaa ttttttagag agagggtagt80341catgagtttt ttacctaaac agtagcaagt ctgccccaat cttgtgaata ggcaatagtc80401ccttgtgaac tccatttgga atcagaacaa ataatgaaac aaaataatag ttaatgcttt80461gaatagataa ctttgtactg aacatattaa ttgacttgat cctcagttga gagctctctc80521actcactgtg ctgtactgct tctctcctga tccaaacagt aacctctcag ctcacctgac80581cagaatttaa gctttggaca tgatacacag aagcttagaa tatgtcagga aaacatgcca80641tatctgcaga tctgaacccc ctcaaagact tagcacatta agatattctg gtgccttttt80701gtatggtccc cattcttttg ggctgtataa tttttctcta aaatttctgc attacatttt80761aatagctttg ttgagataaa attcatatac catataattc atccatttaa aatgtacact80821tcaatggttt ttagtatagt tgcagagttg tgtaatcatc accaaaatct aattttagaa80881tatttttgtc acccaaaaag gaaacttaat acccattaag tagtcagtcc cacccagccc80941tcactgccaa gacaaccatg aatctacttt ccatccctac agattggcta ttccagacat81001tttatataat tggaattatg taatgtggtc tttttgtgac tggcttcttt tattttgtgt81061aaagttttca aggttcattc attttcagta aggttgattt ctatcaaaat tatgttataa81121ttacatatga ctttatttag caaaacagaa aaagatacag attcaactga tagtgtaaat81181gcaatttctt tctactacct gtctattttc attataaatc aatgctggaa ggaaatcttc81241aactaaaact tcaagtcccc aaactacact tctagtcaca aaaatgtgcc taaattatca81301gactaggaac acccccatga gagaaagtgg ctaaatgggg tatcttcctg aatgggcagg81361ccacctctta cctgggtatt ggacaaggcc cctactccag acgattaatc ctccttccca81421ccacaggctc actctttgca ttcttaggca gttatgactc tgtaaaaatg caaaagtttc81481taatgcttca gttgcggtta gctttttgta gttaaaattt tagtaatcag ttcataggct81541aaagctttgc taatcaaagt ctgatttatt gattagcaat atagtataac cagtgaactt81601ctttgaaaat agaatctcaa gactacctca gacctgccta taattagcat tttaagattt81661ccagatgatt catatgcata ttcaaattta agaaccctcc cctaaagagt attagaagaa81721attagattta aaagagatat ctgaggaaga taaatagaaa caatgaatga atgaatgaat81781gaatagaaag atgtagcttt catttgtttt gctttacaat ttgcttttgg ttggtactga81841tttaagtgct ttagtaacta ttaaagtggc attcagggct cattctagag cccaggatca81901taatcactca ttgtgcacaa tatatgccta gtacttcaca cattgtgggt gctctaaatg81961tatgttatct ctcttttcct ttctaagata taaaatttca tagtagtagt attaaaataa82021gaagaaagtt ataatgcaat ggaactacaa cacaataatt ataacgtaat aaataaccca82081attgaaaaat gggctgagaa agaggcagag caaaatagct gaaatgaact ccccagcaat82141cgtcccctta acgccccaca ggataaccga gttgaacaac tgcccaggca agaaagcacc82201ttcataataa ccaaaaatca ggtgatgatc acagtgcctg gttttaacat caagaaaaga82261ggcactgaag gctgggtgcg gtggctcacg cctgtaatcc caccactttg ggaggccgag82321atgggctgtt cacgaggtca ggagatcaag accatcctgg ctaacacggt gaaaccccgt82381ctctactaaa aatacaaaaa attagctggg cgtggtggca ggtgcctgta gtcccagcta82441ctcaggaggc tgaggcagga gaatggtgcg aacctgggag gcggagcttg cagtgagcag82581agatcgtgcc actgcactcc agtctaggtg acagagcgaa actccatctc aaaaaaaaaa82561taaaataaaa taaaaaagga aagaagcact gaagagggtg ggaaagacag tcctgcatta82621cctaaactct tccccacctc cccactagtg gcacagcatg geacagagag agaatccatg82681tgtctgggag agagagagag caaagtgatt gtcagacttt gcattggaac tcaatgatgg82741cctatcatag tgaaaaacaa tacagggcag aattctgcca gcccccatgg agggagcatt82801tagaccagcc tcagccagaa ggtaaccctc cactgcagca gtaggaacct gacttccagc82861tagctctacc attggttgac tacagtgctg gggtcctgaa aaaatttgaa agacaggcct82921cagggactac agtacttggg caagtactga tgctcttctg ggcttgcaac cagtggactt82981ggggtgcacg tgacccagtg agacaccagc ggtggtgacc aagggagtac ttgcatcacc83041catctcccaa ctctgtgcag tgcaacttgg ggagagactt cttttgcttg ggaacgggag83101aaggaagagt atagaggact ttgtcttgca acttgggtaa ccagctcagc tacagtaaac83161taaagcaact agcagactcc ggaagccctc tcctgattcc aggccctggc tactgaacag83221catttctaga cccaccctgg gacagaaggg aacccagtgc gctgaaggga aggacccagg83281cctggaagga ttcaccactt gctgattaaa gagccctcag gccttgaata aacatgagtg83341gtagccaaga aatagtcacc acaggctttg ggcaagaccc agtactatgc tgacttcagg83401tgtgacccag cgcagtctca gctataggaa agtaatttat cctcccttgg cctcaatttt83461ctagttttag aaagtggatt atactatcac ccacaataat agatgtgtgt gtgtgtgtgt83521gtgtgtgtgt aatgtcaata tatgtcaaat gcttagacac tacctagcac ataaacttta83581ggaaagttag ctctcatttt tatttgttct gttttgtttc tgattttact gttatatagt83641tgtaaagaaa aaatgtggta tcctaggtag ggtaaaacat taatgtacaa tttaacttag83701ggaaatttaa tgtccccatt atattgagtc ttctgtttca gagcaaattc tttccattta83761ttcaagtttc tttttcgatc tttcagcaga atattcttct cataaatttg ttaaaattta83821ttcccaggta ttttatcttt tcattgcaat gtgaatgaca ttttctagtt tgttatagtg83881tgtgtatcct ataaacacac tatataaaac atattaaaaa tcaagaaaca ttggttcttg83941atatttaagt ttaaatcatt aaacttactg atttgtttta tcatatggtt tagatgagtc84001ttttggaact tcctggttta caattaaatt ccaatatatg gtgataattt tgcttccctt84061ctgataatta aacatcttgt ttctcttttt cctaattgca gtggttgata ttttcagaac84121aattatcatt tttcaattga attatcattg tactcctgga atgaaacaca attggtcatg84181gtacattatt cccttagctt gctgctaaac ttgttttgct aatattcata agtgaggttg84241gcctatggtt ttcttttttg tattatggca ttagtttcca aggctgccaa taacaaatat84301cacagactga gtgaaatgta tctcactgta cattacagta gagttagatt agaacacaga84361gaccaagatg ccagacatgt gcacacacag agatatgacc atgtcatatc tgctactgct84421accatggccc atctgctact tatcacttgt tcatgtacaa ctatacattt ctgtacaaca84481gaaatatatt ttctcacagt tcagggcaat agaagttcat tatcaaggtg ttggcgaatt84541tgatttcctc tgaggcctct ctccttggtt tgcagatgca ttcgctgtat gttaacatgc84601tcatatctct gtgtgtgcac atgcctggca gcttggtctc tgtgttctaa tctactcttt84661ttatgaggtc acttgtcata ttaattaatg cctaccctaa tgaccttatt ttaacttaat84721tacttcttca aaggctctat ttgtaaatag tcacatcctg aggtattatg gggttagggc84781ttcaacacgt gaatttcagg ggacacaatt ccccatagca attatccttg tcagattttg84841aaataaatgt catgttgaat ctgtaaaaat aatttggaag cttttctctg tctctttctg84901tctctctctt tctctgaatc tgttatgtag cctgatgttg tttttatttc acacaatgca84961tcaatattac cttgtctctg ataagtctcc caaaatttca caggcagaga aacaagtcct85021gtttctgtag gagaccacca atatgcacat ctcagactac cataatttgg atatggaaaa85081catcgcaagg gccaaatatg ccagccgatg cacatcatta taaattattg tttcaatatc85141aatctaggga atgtcactaa tgattgatag gtattataat caccattcat tgtcattaac85201tttataggtt tccatacttg gtttttgtag tcatgtgcag ttccacaatg ccaaaacttg85261atataccttc attttaaaca gttaagtttc taaaagtaaa cctttctttt acacatagtc85321ttaaatatct aagactcaat ttaagaacac tgcatattct gtggcattcg aaaatggttt85381cctcgggagt tgaaaccata aactctatac cccattactg agtgtgccct atgttacaga85441atactcctta ttaaaggtga aaaagcaatc tactactctg caggcttttg gtctggagat85501aaacacaaga tcatagatca tgtattatta tcagttattt taaggtctca gagcaattct85561ttaaacatgt atcaattgta aggagtaata taaaaggacg ctagcttgtc aatttctttt85621tgtcactcag gtctttggca tagactctgt tagctttatt aaacattaag tattgctttc85681aggKtgaggt agaagagcat aagcaagaaa agctttatag gctaataaaa atgtaaatga85741gatgttcatc agcttgctgc cctggaaaat gatggaagaa acattacctg aaaaattaga85801ttcaacRatt attgctaacc aaatcaacct gatatgagaa atccctttgt aaccatttgt85861gttaatttgc atccttatgt caatctttct atgcactgaa tttgcttcca acatcctagg85921tcctctRtac atatatatgt agataatgta tatacacatt tttatatata catagaaata85981tatacacata tttgtataat tcatgttatt aattaacatg tattagctaa acagaaatat86041ttctgttaca tatttgcagc agttggtaag aaacgcctat atattttaat tcgtctgcac86101ttagccttct ttttgcacac catactcctt acaaggaatt tgaagactga gttgtaaatc86161agaatttcta tagcttgtta aagtcacatc ctcagagttt ggaaacctct gagctcacta86221tgtctcagga agacgggaaa ttcttcttct Wcttcttctt ccccttaatt aggaatacat86281tgtaataata atattttcta tgtgaccttt gaatacagta gtaaaataac aagtgggggc86341tacttacttt tctcagggat gattcatcct gggatggtgt ttagacagtg gcctttgagt86401attggatatt ctataaacat cgtgtgagtt gttcttcttg atgactccct ttagtggatc86461tccgaagacc ctttagaagc ttgctgaggg tgggatggag gaagaggcta aagcccccat86521aactgactct cagcaactgt cctctctcac agaatctctc ttttgcggtg acccttaaag86581gcacttggcc catctgctac ttatcacttg ttcatgtaca actatacatt aggccaatat86641attaggttgg tacaaaaata aatgtggttt ttgccattat ttttaatcta aaaaacctgc86701aattactttt gcacaaacct aatagttatt gtaaaggatg gaacttttaa gcaaggcaca86761caaaaccttc taaagtcact ctgagaagca attcagagta gccccttgct gcatctttca86821ttttgcagcc attttactct tttctctgcc ctaaagcaga tctcaggtta ctaactaacc86881catttatgcc agaggttgca catttttttg tgtgaaaaat cagaccttgg cgatgacctt86941gagaagtagg atataaataa ctcccacaag cttagcgttc caataatgga acactaggca87001taaatgggct aactccttca gctctgggct gagtttctca caattactgc agatctccac87061atctgggcag gtcctgacaa tagttaaaga ttttttttct cataatttgt cctgaaagga87121catatattga gctattcagg agacttgaga aaagttccaa gttaaatagg ataaagggca87181atggatacac attttttgtt cagaaatata aagattattc ttagttagag gtgaatatgt87241tccttcactg ttctctgaat tcattctgtc attctgctcc tggacttcct acaatagtac87301cttgacttat aaggcccgtg tccagatctt ttacagtcca cttcattcaa aatgctcctg87361aaaaattcaa gggactcact aacacatggc tttggtaaag gcattctcct gaacaaaaac87421ctttctgacc tcaaaatMca cctagcaagg gatttagtgg tgataatcta gctctcctac87481cttttcagcc ctggatcttt atgctcttta tcttgcaaaa catgctctaa attaaattaa87541ctgctaccca tttgtatgcc ccgtgtttct acaacactat agtctcttta tggttcacct87601tttctaccca tccccaaagc ccagatgaaa tgccactttc atgaaactgt tggatccatc87661tatcctgaag tttttctatg ctctgaacaa ccacaatgtt tttttgtacc tttctaataa87721catgtaacac cacctgtttc atgagcatag ttttatttga ttattttatt atactctttt87781ttgagcacag gaactgcaca ctagaaatca tcatatttct cacaggttcc aatattgtga87841tttttgcata cagtagattt ttaataagtt tctgtcaaat gactaaacta ataattattc87901aagtgtttta ggaattaaac agaatgagga aaattatttc tactctccat aaaagcagaa87961gagattataa tgatacgttt catgaagaaa gtagcttaca aaattacaag caattttcat88021gaataacttc aaacaaatta gtcttaatac ctataaaaag tatcacagcc ttaaaaaaat88081gatgacaatt aagtgatgtc agcagtatgg cagaataggt gtttccattg cttgtcatct88141cacagaaata tcaacttgaa gaactatcca cacataaaaa taccttcaca agagctaaga88201aattcagatg agagattatg gcatctgaat ggagcagaga aatcagcaaa gacacattga88261agaggacaag aatagtttta cattatgcat gtcacccttt cctaaagcct atgcagttca88321gtacaaatag agatagcctc catgtaaagg aaaagaataa agtgagtccc taacttcatt88381gcaggcccca gcaccatgcc taccccagtg gattccagaa ccaggccagt cctcatgtcc88441ccagacctct ctctggcccc catggaccca gcatccaggc ccatgtcgct gccaaactag88501cccctgtagc ctgtggcctc aggcctcagg ctcattctaa tatcagacca gccctcacag88561ctccaggcct gctctgggcc accccagtgc caggaaggcc cccataagtc aggctgcagg88621ccagccctag tagtcccagg ctactaggga ctagagttca tgtaagccct cctacatgaa88681ctcaggtacg aggctcattc atatggaccc aatgtccagg ccctccccag tagacccagg88741ctccaggctt gcccctacaa aaaaagacat cagattgacc cctgtggact taggcactag88801gcccatgaac ccactgacct aggcagcagg ccagcctgcc tgaggactcc cgtggcaagc88861ccactgcaaa ccccaatgga taacctggtc agaatctctg gacaagctgt ctggtgaagg88921gctttccttg ccagtctgta aagactggaa gaagtgtcta cttcaaatgc acagatacca88981atacaaggcc acaaggatca caaataatta gggaaacatg atgctaccca ggaaacaaag89041caccaggaac tgaccctaag gaaatgaaga tttctgaact gcctggcaaa taattaaaaa89101taatcacctt aaagaggctc aatgtgctcc aagagaacac agatatcaga agttcaacaa89161agaggtagaa accataaaaa gaaatcaatc agaaactcag aagctaaaga gcacaatgat89221tgaactaaaa aatttcacag agaacttcaa cacaaacttg atcaagcaga agagagaatc89281agcaagaaaa aaaagaaact aaggaaaaaa acaggtcatt tgaaattact cagataaaca89341aaaaggagaa aaagaatgac aaacgtggaa aagcctacag cacttttggt acatcaacaa89401gtaaaccagt gtatgcatta ttgaggtcca agaagaaaca gagaaagaga aaggggaaga89461aagcttattg aaagatatca tgacagaaaa cttttcaagt ttggaaaggg aaatgaatat89521ccttatccat gaagcccaaa gaaccccaaa gatacagaga tattcactga gacacatcat89581aattaaattc tcagaaatca aagacagaga aagaattttg aaagcaccat gagaaaagtg89641actcatcaca tatataaggg tataagaata tttattcaac aaaggtggca ggaacacaca89701ataggaacag gacattctct tcaataaatg atactgagac aaatcaactc aaaattgact89761aaagacttaa atgttaaggg ctgaaacttt aaaactacta gaagaaaaca tagagaaaag89821cctttttgaa attggtttgg gcaatgattt cttggctatg acctcaaaaa cacaagaaac89881aaagcaaaaa gagattaaca ggattgcatc aaactaaaaa gcattggcat agcaaaagaa89941acaatcaaca gaataaagag acaacttatg aaatgggagg aaatatttgc aaaccataca90001tctgataagg ggttaatact cgaaatatat aaggaattca actcagcaac tcagtaaaag90061gacaacaaat attccaatct aaaaatgtgc aaaaaacctg aaaagacatt tctcaaaaat90121aggaaataca aatcgccaag aagtatatga aaaaaatgct taatatcgtt aattatcagg90181gaaatgcaaa ttaaaactac aatgagatat taccccatac cattataaaa tggctactat90241caaaaaggca aaaaatagca agtattgaca agaatatgca gaaaagtgaa acggtacagc90301agttatggaa aagagtataa agattcctca aaaaattcaa aatatgacta ccatgtgatc90361caacaacctc atttctgggt atatattcaa aagaattaaa atcagcatat tgaagatata90421tctgcattcc cattttttaa aaatggtgtc tcattctatt gctctctcat tcgtattgca90481acattattca caatagccaa gatgtgaaat caaccaatca acctaagtgt ccatctattt90541atgaatggat aaagaaaatg tgggtataga atagtataca tatcatatat gtgaatatta90601tttatcttaa aatgaagaaa attctctcat ttgcagcaac atgaatgaac tgggaggaca90661ttatgccaag tgaaataagc taggcacaga aagacaaatg cttcatgatc tcacttacat90721gtaaaatctt aaagccttaa aattacagaa gcagagagta gaatggtggt tgccgggggc90781tggatgtgaa gaaacaggga tgcgttggtc aaaggatacg aagttttagt tatgaagaat90841gaataaattc tggagatcta atgtacaatg tagtgtttat aattattaat attatgttgt90901atactttaaa tttgctaaga gagtaaatct taaatgttct taccacaaaa atttaactat90961gtgaggtgat ggatatgatc attaggttgg ttgtggtaat cacttcacaa tgtatatata91021aatcagtcaa agcatcatgt tgcacaaaat aaatacatac tttttatttg tcagttatac91081ctcaataaag gtggaaaaat aaatttttaa acactgaata taggtaaagc taaaagttat91141atgtggggat atggggaaag agtgtatgtt gtggcttgga gcaacaagta aaaggattac91201atcttcatct tccacatagt gtggttaaac agaaaaaaaa taaatattaa aaaaatcaat91261tatttttaca ttgtgtatat aaatatccaa agaagcagct aaattagtca aagtgcttat91321ctttgggaaa taataatgga aggagaagta aaccagagaa ctttgttttt caaagttgac91381cttgtagaac catttttgac ttaaaaaatg gatatatgct gttttgattt ttaaaaatta91441attaaaaaat gatgatcaca ttttgtcttt aaaaattttt gtttaaaacg tatccatttc91501ttctccccat ttaatatatc ttttgaccat ttaaagtata aaaaaagaat ttggacttaa91561tctaatgaag aagtttgacc ctaacaaaag agagtaatgg ggtttaaatt tatgccactc91621tacaaaaaag aagtcgaact cacccatagg tagtgtcatt agtcttttag gtctgcatgc91681ctctctctat gacttttccc aatctttgtt ttaatcaata atcttgtctc cagtctaaca91741tgggaattgt gctctctggc ataaaatggg ctactgaatt acagtcctgt gctatatcac91801acagccttgc cattgatagc atggaaatac ttcgaaatac tgctgtcaaa atttgcaaac91861tgtcaagata aataagttca ttcagattat ttatatctgc tgtttctccc cctaacctca91921tacttagaaa attatatgtg tctagtcctt ctccaacttg attttatgtc ttttagccta91981atacattggc aaaccaaatg catgactctg acaccgataa tactaagaag ggctgaaatg92041aattgttaga atgagagaca gtcactttgt ttagaaggga gtgctccatg agcagtgtga92101taacatagct ccttgaaaat ctagtcaatc accattttat agtaaaagcc tatgaatttt92161tgtagcttgg tgtcttattg tctctattca atagttgctt ctaacttttc tgggcatccc92221cagaaaagtg gaaacctcct gtgaatgagg caaatttagt aacctccaat taatcaagca92281cactaagcac ataccacatc tgcaggaata aactaggtat gggattaaag agatgtatag92341aattaattct ttgcttaatg tattttgcca ttctatattt cttttttttt tttcaagaga92401gcaagtctaa gtcctccctg gagaattgtt gaaataatgc ctaatttagg taactaagca92461ttttaagcct ttctacactt ttataatatt tgtaattgaa aaaaacccag caccaatata92521taaataatta atttaccttt gaaatactgc tgtcagaatt atgatgcaaa ctctgcatca92581catttttgca taccttttaa cccaggaaat tcacttctag aaatttattt tacgaaaata92641ataagagatt gaattgcatg ggtgttcatg caaaataaaa aagtaaaatg ctctaaatgc92701ccagcattaa atattgatta aatcaaaaca gtgcatggat atggtgggac actatgctat92761cattaaacat gacattgtac aagattatac attgacatga aaagacacat ataaagactt92821tgtgaaacat gtataatgtt atttaagctt tgttgtgaca ctcaagtttt ttctttcttt92881ttagtttttc ttattactag aatgaatgca ttttcttaaa tattcacctt tacagaagtg92941actacacaat taatggctac acacctctcc ccacttattt ctgcccaaag acaactactg93001ttgacatttt ggtggatatt ctcctaggtt gtttcatgta tgtctttttc aaatgtagga93061tcacactaaa tgcttctgaa gcatactttt cttatttaaa aaatacagca gacagcttct93121aagtgtgcac catgttcttt tgtaaacata tgtattttaa atatatgcac acacataaag93181agagagaaag aggctgttta aggagtaatt gttaagtgga tatgcatgca acgaccacca93241gtttaagaag taatttctag ttccccggag aatcttcatg agaacttctg atttcatatc93301tgtctccttc ttctagagta atccctctct ctctatggta actattcctt tgctttccta93361gatattaagt atatgtaaat gtatgtgtaa ctctgtgaat atgtatctct gtataaaatc93421attaattaaa tatttttaat tttttctgtt cttgaacttt atgtaaatga aatcaaaggc93481atatttttgt gactttatta tttattgtga cttctttgtg agtgatcact tatgacattt93541taacaatttt tttatgcatg tggctgttac atagccacat aatatatgtt tacataggtc93601ttatgtacaa gggaaggttc taagcacatt atacagatta tcttgtttta tgtcgggaat93661aggtatcatt attcgtttca ttttacagat tgggaaactg agttccagaa aggttaaata93721acttaatgta ggtcacacag ctagtaagtg gtgtagatta tattctatcc taagttgact93781gacttcatag tctgtgatct taaccatagt tccaacctgc tttctgttac agatgagtat93841gctacagatg ccttctgctt atttagggtt aatgctttca tatattaaga atcccccaag93901actcaatttc ataaagatat tttaaattat tatcttaagg tttgatggtt tgccttcagt93961atttaagtgc tttacctacc taatattgat tccagtctat ggtacaaagc atgggtttaa94021cttaattttt tgacatgttc aattgtctca acacctttta ttaaaaagct tatattttcc94081ttattaataa tatacaatac taattctgtc atatatcaag tgtccattta attgcaaatc94141tgtttctaga tctctgttct gttcatttat ctacttatct agtcatgcaa catcctatat94201aaagttgttc agaatagtct cttaataatt ttaattttct accaatagtg gcatattctt94261ttttattcct gatactgatt attttggtct ctctttttgt cggtcttatc agaaatcaga94321agtttagcaa ttttgtgtct ttgcaaagaa ccaacttctg gctttgttga atttctctag94381tattatatgt ttgtcttttt tttaatcaat ttctgctttt atctttctta acttcatata94441atttcactac tgtttttcta atttcttaag atggatgcat agcttagctt attaatattt94501agcctttttt ttctaacagc attttaggct acacatttgt aagttctgtt tttggcacat94561ttaagttgta atatattttg ttactgttca gtttaaaaca tttttccttc tactagaaat94621tctttttgga cccataaatt atttagaagt aggtttccaa ctttacaaac ttagtgtttt94681taaattacct tgatattgat ttttaagtta gttgtactgt aagcaaagaa cttcgtcatt94741agttttccag tatttgaaat gcattgagtg ctgctgtaag ggccaatatt tagtcaattt94801tgtgaatgtt ccctttattc ttgaagtgaa tatgcattct gcagttgttt gctacacagc94861ttataggtac aaatttgatc aaggtcttaa ttgtgtcctt catgtctttc atatctttat94921tgtttcttgt tggctttatc aattttgtgt taaaaatttt tcaccatgat tgtagatctt94981gctgtttatt gttgtagttc tataaggttt tgcctaatat attttgatgY tatgtgttat95041tgatataaaa tttaacatgg tttatagttt cccagggtga attaatgaat acattacaat95101agattgaact tctctatctc tagcaatggt ttactttacc catgaagtcc aatttttccc95161ttatcaattt aataatatgt gactttattg cttagaattt attggacata tctttttcta95221tactgttatt ttaagacttc ataactgttt tactttaaag gcattttgta aatggcatat95281aattgaattt tattctaccc tagacagata ttcttttgcc tgaaagatta attcactttt95341attagatatt attacttaca tatttggagt tgaatctacc aacgtatttt gtatttccat95401ttgctccatc ttgtcttacg tcaaatatta agtgcctgta tatgtgatga tctgtttcta95461gactctatat taacctactg ttctatttgt ttgttgtatg tgagtaccac tgtgttttcc95521ttgctgaagc tttacaatga gttttgacat ccaagcatat gaatgtaata atcttcttct95581ttaagatttt ctttgttact ttttcgcttt ttacatttta ttatatatta aaatcagatt95641ggtaattctg tgatttttga tagaactatt ttttaaattt aaattaaaat tttcatttgt95701ttcttgttgc taatgaataa aaatataatt tttgtacact ggccttgttt ctagtgattt95761tgtttaattc acattttaat gatttatatt taccatggga atgattcgat tgcaaataga95821ttcttctgtc cttagttttc tatttctttt tctttcctta tttcattggc actctagaac95881aatgtcaaac agaaatggtg acaaagaata gacctgtttt attcctgatt ttagggataa95941aattatattt cagcattagc tatagagttt ctataatttt gtgattattt tctaagtatg96001tatagacatt tttatcacat taagaatttt ttccttctat tgtttgctaa gaatttagtc96061cacaattact attaacatta aattgttatc aaatattttc tcatctattg agatcattat96121gtaattttct tctattattt cattaatatg ctaatttaca ttggttttga atgccaaatt96181aattttggat tcctaacata agttgcactt tttctgacat gtcattttta tactcgttga96241attcagattg ttgatgtttt tattttttaa tttacaaata aaaattatat atatttatgg96301tgtacaacat gatgttttga tagattcttg atattttctt tagaattttt gcaactgtat96361ttgaaagaca ttgtcctata atttattttc tcataatgtt agaaaaccct ggcctcatag96421aacaaattag aaagttctat tctttggaaa agtttgtgaa gatttatgtt atttcctcca96481taaatattta ataacattat ctagtaaatt tatetaggat atgggttttc tttctggaaa96541tgtgttcaaa ttatgaattt gatattttca gaagatatgg atagtggttt gtgggaatct96601gttgatactg acactggctc agggtgcatt ttgcacacct ctttctaact ctctcctcag96661tgatgtcatg caagtagctt aaaattagcc attgtggaaa tatttacaac acaaaaatca96721aaaaaagtgt ggcttatttt ttgagagctg aaggaaaact taaatgtttt cacttcttcc96781tatgttcatt ttttaaagtt gtgatttcca aggcatttga ttatttcacc caaatctcca96841aatttattgg cataaatttg atcataagtg cttcattata tttaatgatc tgcaataatg96901atctttttca ttactaatat tatgtatttg ttccttctca atatttttcc tcaggtatct96961ggtcagaaat ttttcaattt gtaattattg tttgtttaaa aaccttttgt gcttttgtta97021tcttctctat tttttttact caatttcatt gttttcttct ttttaactgt ttcttttagt97081tttctgggga aaatataact tggaattttt tctaacttct taagatgaat tattagatta97141tttattttca tccttttttt attgtttaat atttRcattt aagactatag tttttcattt97201aaatacaggg ttagtggtat acctgaattt ttatatattt acacactcat tcacttcaaa97261tattttcaga tttccattat tatttgtctc atgggtcatt tattttttca ttgctaaatt97321ttcaaacatt ttgacagtta ctaattattt ttttactttg atttgcagta taattgcatt97381atgctcagaa attatagttt


Following is a PCLO genomic nucleotide sequence (SEQ ID NO: 6).

>7:82022801-821125001agttgcctcc ctttgccccc ccacccccca acaggccctg atgtgtgttg ttcccctctc61tgtgtccatg tgttctcaat gagtaactgc catttctgct aatgcatgta attacatttg121tgttcttatg atatatattg gttttcatcc agggttcctg gtttataact cccacagccc181ttattacagt cttttgttac aatgttgggt gtgtgaggcy tcaggggcag gcctcaggaa241atagaatctc tctcctgccc tcctttcagc tgccccaagg cagaactcta atcttctccc301acctttctga tgtgagtctt aagaccctcc ccagggaagg tcccaccctg tactctgtgg361gaagaaatgt tgacatcatg aagcttccat aaaaacccaa gaggattggg tttgaggagc421ttctggagag ctgaacacat ggaggttcct ggagggtggc atgcccaggg tgggcatgga481agcttctcgc cccttccccc ataccttgcc ctgcacatct cttcatctgt gtccttgata541gtaaacctgt aaacataagg tttccctgag ttctgtgaat tgtttcagca aattaatcat601acccacagag gagatcacag gaaccccaac ttgaagccag tcggtcagaa ctttctaagg661cccagacttg tgacaatact tgtgtgtatt gggaggcagt cttggggact gagcccttga721cctgtgggtt ctgacactag ctccaagtag atagtgctgg aattaaattg gaggacaccc781agctggtatc tgtaatttgg tgtgtgggga aaacagcccc ctaacctctg acgtttggtc841acagaaatct ctgttgcttg ttgtggtgtg agaattgtgg aaaaacatga tttgaggaga901ggtttcttga aataacattc ccttcctttt ccagaaaaaa atcttctata cataaatgct961cagttacagc atgcaaacta ctaataattt atttcattat tttcatttat tttgcttaaa1021tcacagggac tggctatgtc tgttttagag gactgatttt acagtgcatc caatattgta1081aatctactgt ggcaacatat gcttagccag aagaagtcga aagagaagag agtagaatgt1141tctattctgg aaatagtttt catgtcacat ataaaaatga tatactaaat gaattatgta1201taaattctaa atgagctttg aaactttttt acgttaatat aaaatagtat agaatatcaa1261aatccatttc agaaatttac tacggttcac tgttatgttt ttcatttcaa gcaaaactct1321tctaaagttt ccataatata aatatatttg aaagtgtctt atatRttagt atatacagag1381atgccttaac acaatctctt attttatctg tctagtatct tttctcctct ttcaccacta1441tctcaagaac atctactccc aaatgcatat ggttctctta agctggaaac caccatactc1501aattttcttt gtcacgatgt ttgttccagg taagtgcatc tgacccacat aggaaaaacc1561agagtgcttc actgaggatg ttagacatat tttctccttt gggtcaagag tgtaaaacta1621ggcctcaaag gagccagcaa cttcatccct tgccttatag aacaacctga gaaaaggcca1681actttcagag aaaagctgag aaaagatgtg agaactaaac aggcttgagg ctggtctttc1741ctgagactat gatatctaca tgtgaatttc ttagtatggc caccagtaaa cattctttgg1801ttttgagttt gtttgtgtag ttttccatct cttaccagtg aaaatactgc cttatcttag1861agttaaagaa atatcacatt taaaaattgc accctagagg gccgggtgtg gtggcttacg1921cctgtaatcc cagccctttg ggaggccgag gcgagcggat cacgacgtca ggagatcgag1981accatcccgg ctaacacggt gaagccccgt ctccactaaa aatacattaa aaaagaaaat2041tagccgggca tggtggcggg ctcctgtagt gccagctact caggaggctg aggcaggaga2101atggcgtgaa cccgggaggc ggagcttgca gtgagctgag atcgcaccac tgaactccag2161cctgggcgat agagcgagac tcggtctcaa aaccaaccaa ccaaacaaaa attgcaccct2221agatgtaatg aaaaatttaa tgaatgttaa aattattcaa tattgttttt aaattaatta2281aaaataattt aggcataaga gaacataaag aatatatcat aagaactcat gcaatggctg2341gtcagcttaa gaaatcaaac aaattcagat gaagccaata gggtaatcct ttcccaattg2401cttctctctc tcttacccct gaagtaatca ttattaactt gattattata attaccttgt2461tattttaatt atatataatt atatatgtat agttatatgt attacatgta tgtatatata2521gacacataca tacatgaatc cggaagaaat aggcaaattc ttagcccttc tcactgtatt2581aaatgataaa cctgatacag acatcatcat ttctttcctg aataattgca acaaatattc2641ggaggtcttt cttctttgac cctttttctc ttcagaatat tctcagaaga gtaattagga2701tcatcctttt aaaacttgag ccagactgtt tcattcctca cctcaaaacc ctccaatact2761gtcccacacc atgttgtgag agttcttgca gtgttcaaca aggcttctat tctctctctg2821acctcatcac ctaccattct ctccactgct cacttcacct gggtcatact cttcctgaag2881gtgctgggca cacttctgct ggtcagccgg gaacgttatt tgtaggattc acacagctct2941ctccctcatt ttcttcagtg attacccaaa tgtcattttc ttgtgaggcc tttccggata3001atattatttc caagattttt tttctttttt aattgcagat gcccagaatt gaattgcccg3061tattcatcaa ttcacagcac acaaccacac aattaacatc tgtctctgcc agaacctgta3121tttgtttggt actttgttta gtttatattt ttccctttta tcttcctaac tttatcatcc3181tcattaccct cccaaccaaa ggcaaccagt cttgagtatt tactgtattt cttttcaatt3241caacttctat gtatttgttt acatgaatgt ttacaaaaag gtatatctgt gagaatgtct3301agtttggctt tctatctatt tattctgaac taaaataata tcatgctgtg aatctcatta3361tttgtactca tttatttctc aaatctatat ttttgaaatc tattgatttt attatatgta3421gatctagata atttttttct gacagttgca tggtattcca ttacctgcct ataatatatt3481ttatgtatct gttcctctag ttatggagaa ataaatatat tgcctccagc tttttacaac3541cacaaaaaac tggaaaacat ttctttacta atgcctttgt aaatgtttac ttaccaaata3601atttaagttt ctttggtatt attttatatt tcattttttc ctttagcatc acaaaacaat3661tattctttat tattccagga cagtggtcct caaccttttt ggcagtagag actggttttg3721tggaagacaa ttattccatg gattgggggc tgtgggggat ggtttcagga tgaaactgtt3781tcatctcaga tcatcaggca ttagattctc ataaggagca cacaacctag atccatcaca3841tgcgcagttc acaataggat ttgcattcct aagagaatct aatgccactg ctgatctgac3901aggacgcaga actcaggagg tgatgctcgc ttggctgcca ctcacctcct actgtgtggc3961ctggtttcta acaggccaca gaccaataca ggtctgtggt cctagggttg gggacccctg4021tactagaaga aataaagctt tcaaagttat aagctaagaa attattctat atatttctat4081agcccactat cttcccatct cccaatcatt tcagctcata gttttgcaaa agagatagaa4141cttaaaaaga tcattacaat tacagggtga ttggcgcaat gatggaggca ctgacaggag4201caagagaaaa cttccttaac agagccagat ggagagcgcc cacagatagg ttcttgaaaa4261agtaagctga ggggtgggtc tcaaaggatg aaaaggtgtt ttccaggatc aagcattttg4321ggaatgggtg atcatattaa gcaaagagaa cagcaggagc aaacttaaca agagaaactg4381cagagcctgt gtggaagact caacacaatt ttgatataac cagatgtaaa ttgggagaaa4441ggacatgtca ggagataaac tggagaagta gaaagagggg ctggttgtgg taaaacctag4501ctagagttat atggttcatg aggtccacag aaatatttta tgctagagag tgacaaagtc4561agattagata gtggctgtaa aagagatgtg aagatgatga taattcatct tcaaaaactt4621ctaaggtaga ttaggagatc agtgtggaca ctaggcattt cttttggtga aatgtgatgg4681aaacctaaag ttaggcagta gttagaatga aaaaaaggaa agagatttaa gaaatcctga4741agaggtaaaa tctgcaagga tttggcaatt gcctaaccgt ggaaatgaga cagtgattag4801aagaataatg taactaaaac caaacctgag atataaaata caggaggaga agaaatttac4861aggttaagag ccatagtatt ttcttctatc ctctagcaaa acaaacaaac aaaactacaa4921tctgatatat ttttctatac caataaattt gcatatttag agttttaaat cttaaaagtt4981tgtttaaatc gttaacaaat tattcttaat tagttaaaca attatgttta aggaacataa5041aattgtatat ttccatatct ccatattatc atgtttgcca ttacaacaat tcaacctaga5101ttgattctag atcgtatttt gcaaaatgaa taattagtga ctcattaatt aataaagaca5161caactttttg ttttgatact ttaagttctg aggtacatgt gcagagcatg caggtttgtt5221ataggtatac acgtgccatg gtggtttgct gcactcatca acctgtcatc tacattaggt5281atttctccta atgctatccc tcccctagct ccccaccccc caacaggccc tggtgtgtga5341tgttctccac cctgtgtaca tgtgttctca ttgttcatct cccacttatg agtgagaaca5401tacggtgttt ggttttctgt ccttgtgtta gtttgctgag aatcatggtt tccagcttca5461tccatgtccc tgcaaaggac atgaactcat ccttttttat ggctgtatag tattccatgg5521tgtatatgtg ccacgttttc tttatccagt ctatcactga taggcgtttg ggttggttcc5581aagtcttttc tattgtgaac acacaataaa catacgtgtg catgtgtctt tataggagaa5641tgatttataa ttctttgggt atatacccag taatgggatt gctgggtcaa atggtatttc5701tagttctaga tccttgagga ataaccacac tgtcttccac aatggttgaa ctaatttaca5761ctcccaccaa gactgtaaaa gcattcctac ttctccaYat cctctccagc atctgttgtt5821tcctgacttt ttaatgatag ccattctaac tggcatgaga tggtatctca ttgtggtttt5881gatttacatt tctctaatga ccagtgatga tgagtttttc ttcatgtgtt tattggctgc5941ataaatatct tcttttgaga agaagacaca acttttaaaa gcaggttttt aaataactca6001ataatttcat ctcattctat tgttttaacc catgccacaa ttatttattt ccaggagcgt6061tttagtagaa tattttattt aaaatatatt taaatacttt attaagaaaa ctatgaacaa6121gaaaatgatt aactttagtt ctacaaaagt ttaaaatatc acatgacgtg ctagacaaat6181ctcaatctga gactgctaat ccatgttcct aaagtcattg gaaaatgaca ttctggaaat6241agatcatatt cagagctcta aaacacccat aaaccaatga gcaacagctc atattaattt6301tactcagata aactcacggc tgactattta aaaaaaccct gtaaattggc tgaggaataa6361cacagcagtc tcaacatcaa actaaattgc aaaatttgac ctaaattttg acctagccaa6421gtcatcatta attaaaagga aatgaactca aactatcaga ctatcaaaag taataaacac6481ttgaagaaaa agctgtcttg aaaataaatt gttgagaaat agttcctgtt catttggcct6541cttgcagtaa ctgaagtcat tataaactac cttttccacc aactaaaata gaagtcaaat6601agagcatgcc tctggttagc taaacaczaga accatagaat gtgaaatgga ctcaggcact6661tcagcactaa aagatggctt caaccaataa gtgtatttat ctgatttaat ttaatcaatc6721tgatttaaat ttataattta tatatttatg atttaaattt aatatatatt taatatataa6781ttaataatta tatataatat ataatttata tataattaat aatttatatt agtatataaa6841tttaatatat ataattgata tagttatgat ttaaacttaa taatctgatt taaaaagcat6901taatgaatga aataaatatt ttttaaaaac caaaataata actcagttaa ctgccttact6961ctagaagatg tagtattttg ataaattaca tttcttctgg ttcaagaaat tccttaatgt7021tctcagcgaa ctttgtgaaa tgcatatagc aacatttaag tcttcccaaa atagaattgt7081aatttctttt gattaagcct gcttttcata ccttatggta gcattctgaa tttctgatct7141ctttgtataa taccatagat gcataaaata aaaagtatta aactaattct cctactattt7201tcaggatatg cccaaaatgt tttaattaaa aacgtcaaat ccttcttcta gacttgtctg7261ccagagattt tctgccttta ctgagagtgc tccacacaca cactcacata ctaatcagaa7321tgtggaagag tcgaattgaa tcaattcttt aagtaacata aaattgtgta tttccataac7381acgtctccat gttatcatgt ttgccgttgt aacaattcaa cctagactga tactagataa7441tatttctgca aaataaatag tgactcatta attaataaag acaaatttta aaagcaggtt7501ttaaaataac tcaatcattc tatctcattc tgttctttta atctgtatca caattatttt7561caggaggatt ttagtggaat attttattta aaatatatct aaatacttta ttaagaaaac7621tatgaaaaaa aatgattagt tatataaaag tttaaaatat ataatgaaat gctagacaaa7681tctcaatctg agactgttaa tccatgttcc taaagtcaca atcctaaagt cacaagaaaa7741ccacattctg aaaatacata atattcagag ctctaaaaca cccataaatc aatgagcaac7801agctcatatt aattttactc agataagctt gcttgtaact gtaaaaaata gttgaatcaa7861ttagattgat tcaattagat tcttaaatcg aagaagcatc aattagaata acctaattga7921attagagtct tccatatttg aaacaacagg tcaatgtcat tatctatata aagctaagaa7981agcaaaatgt ggccaggcac agtggctcat gcctctaatc ccagcatttt gggagactga8041ggcgggtgga tcacctgagg tcaggagttt gagaccagcc tggccaacat ggtgaaaccc8101catctctact aaaaatacaa aagttaactg ggcgtggtgg caggcacctg taatccagct8161actcgggagg ctgaggcagg agaatcactt gaagctggaa ggtggaggtt gcagtgagtc8221gagatcgcac cactgcgctc cagcctgggc aataaagtga gattctgtct caaaaaaaga8281gagcaaaatg caaacattct acctaattca aactccaaat tactctttta actcatcttt8341aaagtactat attgtcttat aaagcatagt caatttattt cttcaaaatc ccgtgaaggt8401ttgcttttca tcatgttcga acagtttccc taacttttaa aatcagtttt tacattgccc8461tctgctgtta acatttcttt tctgtttaat ctttctaaat catacttgtg tttagtgatc8521attttacatg atgaccatta tcacatcatt tcatctttat gatgctggtg cccgaagctg8581ttttatttgc attttgattc tgaatcattt ttagaacaaa gaacctcatt tgtcttccta8641gatcactctc agagctgtct gtcctgcttt tcgcacctct gcccctgctt gcattccagt8701ccttatctat tgaattgcct ccaaacgagt cccccaactt cagataagat cctctctaac8761ccatccttca gaaccccaaa tctgtgtaat tttccttctt aaaatcaatc tatatcttcc8821aattgcctat ttacagtgtg gagtctaagt tcttcctgac agcttcagat atttcaYaat8881ttatctgaac tgtgactgct ctcaaatcct atcttgcact actccctcca gcgcagaact8941atgctacagt caggcaactt gtccacaagt tcaccaatat cacgtatttt ctcatgttgc9001aaaggctttg acaatgatgt ttccccatat tggaatcctt gccagcattc cttctactat9061gaatacatat tacttttatt tcaagaacta tttaaaaaaa tattcctgcc aggaaggctt9121ttcttatcct tcctacagca gagtacctta atattcaata tacattcaca tctgtcagaa9181ctctatgcca attgttcatg aacatgtttg tctgtcctac tacactggtt ctttttggca9241gagacactgc attaataatc ttctgtagat ccagctgcgc ttgtgaaaag agaagcacag9301tcaatataaa ttcagagtta ctatactttt aagtaaatta tgtagttttt aattgaagta9361ataatctctc tgagttcagt tttagggtgg taaactcctt aaaacctgta taacaacata9421caggattaac aagacttttc aaggtgtctt aggtatggca ggcatatgca aaaatctaat9481gttttaacac actctctcaa tgtttttgtg taaacattta tttcagtttc gcaattatgt9541tattaaggtt ggtttagcct ttggtttctc tgagttttta ttacttgatt caaacatatt9601tatcaagtgg gcactttgtg caagtccctg tgccagacac tgggaatagc ataaatacgg9661taatacctta ttataaagga ttcctgtgag aattgggttc accagatttt ttaaaggacc9721tagccaaatg cttggcaaat aaatgaaagt tgtgttgttg ttattgttat taaggggttt9781gtgtagattt ttttaaaaaa gatgaatcat caattagaat gaatataaat aatcattcaa9841tatgacaata tcataataat atgtgtaaga taaagccctg ctactgctac ttagcattcc9901ataacaccaa caatagttgt ctgggctcca gagtgcaatg taaattagag aaatcacaat9961ggtgtgctat ttcttggata agtgagcatc cactttggat gataacagcc taaaattgca10021tctgcttagt gcaggacaaa acatgacttg agtagatagc aagtcctttt tcaaagtaaa10081agatttaaac agaataatcc acctccacag tttttagtgg tacaaagcaa ataatagaat10141atattgatga ttttagagat tagcagtcaa catttaataa acaagctgtg ttgtgctttc10201ctcagagtct attatctcgt gtccatagta ctttcttatc tgtaaaactg gttccttaag10261ggaaagtaat acttggttga ccaagggctt cttttctgaa atgtgctggt cttttctttt10321tccagtttta cttttttact tacctataaa atattttctt atacagaaaa ttatgtaaac10381ttccaggatt ttctagatat aataaatttt aaaaaatttt cattaaagta ataaactaaa10441aggtgggaaa gtgaattatc ttcaataaat tttgtcattt ttaaaaattc tgaatttttt10501cttcctgaac atttataaat ttatttttaa actttcagtg aactggaaaa agaagttaaa10561agttaaaaac tttgaatttc atagactttt tttcctccag agattttgta ttttacatca10621tcaaagtcag gttttttaaa agtgaaaaaa actttctcta ttcaacaatg aagctatcag10681gaggtcttta atgagtgtgt cagataaaaa gagtacttaa ttgtaatacc tactactgaa10741gagttttaag agaatggata ttcataataa tttaggggaa tggtagaaaa ttatttactt10801caaataatac catttttttc ttcttattat tatgcagttt gattcctatt gcttgttttt10861catttcttca tataacaaaa ggttgaataa ttttatggct cttttttttt tttgacaagg10921tctcactctg tcatccaggc tggggtgcag tggcatgatc tcagctcact gtaacctctg10981cctcctgagc ccaagcgata ctccctcctc agcatcccaa gtagctggga ccacaggcat11041ggcgccacca cacccagcta attttcgtat tttttttaga gatggggttt tgtcatgttg11101cctaggcttc tcttgaactc ttggactcaa gcaatctgcc cactccagcc tcccaaagtg11161ctgggattac agggtgagcc acctcacccc accaccattt tatatttttg acttcttttt11221taaaaaaata aaatctcaca tctcacatat ctcaacccgt tattttaccg aactcatagt11281caaattgcta agagaaaccc actgccaatt atttttttta aatctgtttc ctgggttctt11341tacattcata tttcttaaaa taattttgta ttgcctttat aatatatggg ctttttaaat11401tttggtatat gaagatttag cttttctcat ccttgtcata tattacgatc ttggttaaac11461taacatttgg aatttatatt aacttgcctc tttaatattc atttttgaga aaatttggtt11521cttatatggc aagctttact taattttctt tttttcctta agttattaat tgcttttttt11581ttttacaatt cgctcagctt tctgcgtacc tatctctaac tgttctcaac acttccaaga11641gcacctcttt caatactatt tttcacttgg ccagacacat catgaatttt gtcaatgctc11701ttctctcctg catgagcacc ccattcactg cccacgtgca gcctagaact ctccatatat11761ttttgttttt gaaattcgtg ttatggtgga gatgtcaaag acaaggaaga gtcacattct11821atgccagtgc tatccctggc tttgaccttc ccacctgggg tttttaaggg tagtacctat11881gtgtgtggcc gacgctcagg tgaagatgca aagcagtatc agctctggtt ttgagtccca11941gccaagttga tggacactct gccgcagttg cacccatatg ttgagaaaac aaaactacag12001gctggctctt ctgctagtgc ttggtcacag tgtataccag ttcacctccc tggagacaaa12061cgccattcga aattgggttt tcagggccct ggaagtattg aattaatctc taaacttctc12121tagtttctgt ggagggccaa ttttttttat tgggctacta gaaatgtctg taccccaaaa12181tacatgggaa tcaaattcag atttttaata actgctaatg aactcttcat atttgatatt12241tgatctccac atttgatatt tgatttccat atctctatat ttgataagat tttctcaggt12301gttctttatg cctgctgcat ggctattgtc tttagattct ccttatctca tcatggaagt12361cctctattct ttgtgtctct ggattcacaa agtattgtgt gtggggtcag ggtatgcttc12421tatgtggtta attcgctatt ttggtggagc atatcttcaa ggagcttcct gagataggta12481gcatggaaag taaaatgttt gtacttagcc ttctcaaaaa tcatttcctt ttacccttac12541acttgatgga tagtttgtgt tagattttta gtttacaatc ccttccattt aaaatttcaa12601aggcattaat tattatcctt ttctttttaa gcctctatta ttgctgttga gatatcgttt12661gaattcttat cttttcttct ttgtagttaa cgtacattaa ttaattatct ggagaaacct12721ttagcatctt ctctacgttt tctgttttct gaattattat gatgtctcac gctgcaagtt12781tatttatttc ttgtctttct aatgtggaga tttgttttct ttagtttaaa gaaatgtgtt12841gtattttgaa taacgtttct tttcatctct cccttttctg tggaattact gctagttaga12901atttgagcct gataggttaa tcacttaatt gttttttaat aattataagt ttttttgttg12961ttattgctaa tttctcttca gtgaaaagcc attgttactg acatgctagt tcataaattt13021tttttctttt tttttttttt tttttttttt ttttttgaga cacagtcttg ctttgtcatc13081caggctagag tgcagtggca cgacctcggc tcactgcacc ctctgcctcc tgggctcaag13141cgatgccccc tatcttagcc tccctagcag ctggaaccac aggcatgcgc taccatgccc13201agctaatttt tgtatttttt gtagagacgg ggtttatgcc atgttgccca ggttggcctt13261gaactcctga gttgaagcaa tctgttctcc tcggtctccc aaagtgctag gactagagac13321atgagccact gcgcctggct ctccagttta taatttttga ccttgtttgt tcttgtttgt13381ccttttttga gcaaaacatg ggagatcctt atttagtata gtgatcattt agtgatcata13441caagttacaa tattttaatc ttatttatca tttgctcatt gcttttattt ttggtttatt13501ttactatatg tgttattatt atttatagac aaaaatgttt attatttctt tgaagcctta13561tggcttctta tcttgcttag gaaattgccc ctcatattcc actaagcctg tacaaatctt13621acttggattt ccttcaaaca tttttattat tccattttta aatttaatct ccttaattaa13681cctgaaaatt atttggtata tggtaatgac tagaagtcta attttctttt cttactgaag13741aaaattatgg cagaactatt tgataaacca actttttcta tggaatgaaa tatctgttcc13801attggaattt ataataatca tttactttaa aaccatatat ggactggtta agttatccat13861ttgcccattt atctatttgt ctattttgtg taatttaatg ttgttttgaa tacagaaatt13921tttaatatct gataaaacaa gatttttaat tttggcaagc tacatagcac ctgtaatgta13981aatggtgtta ttagttgatt gcctatcatc attaagtatt ggatgttttc ctgcccaaac14041caattaagat agtcttatga atttaaaatg ttttttcatg aaatagctct gtaaccttca14101atatcaaaat aaaaatgttg gtaagtacaa tggtaagagt caggtgatgt tagattagct14161atctgtagat tctaactctt tttaataagt tgactataaa actctgcttt gaatggtttt14221actgctctag ttctctgttt tcttgactat ataaggtgga aattattatt tcaattattc14281catatctata tctcctactg tcttgtgaca aatgaaccag acaatgtgaa ggcatttaga14341acaaaataaa cattatatat ttgtgatatt tacacatctt ttgttaattt tctacatcta14401tactaagaaa atattcttgg cttatattcg ttgtgttatt tgctaagttt tagtattctt14461atcttgattc aaaagcctct ctctcataag taaatcagag caacaactcc cagtgcaaca14521tttcagagac ttctaggtaa agaagacatt tcaaaaaata ttgaaaacta ctatttcaga14581tattaatctc tgaaagtcta aaatattctg ggctttcatc aagcaattga caatgctttg14641atgtgtgcta agacgaagaa atacccaaga tcgcttccag aaatttaatt catttaactt14701tttagtggtc aatcgatacc ctccagagat gaatctggct tagatttaag acacagattt14761ttaaaaaata tttcaataat agggtatttt gcagtaaatg ccattaattt taaattccca14821gtgaataatc caggactaga aataagtgac actggatttt agattcacgt aatttaggaa14881aatgtgataa ggtgataagg tccagggaaa tcacacccca aataggagga ggctttcccc14941ttgcaccctt tgctctagct tcactgaata acttctcttc tatatacccc caatcttttc15001ttccatctgt gatgttcctc atggtgttct tcagtctgaa cttggcctca ttcattgcca15061catgtctaga ccttgttcaa ctttcaaagc agtcactact tgtaacctct ttcacaaaac15121catgctcccc accatcactt ctacagatgc tgccatgctt ttcaatgctg aaattccaaa15181gcatataatt ttccctctgg cttatatatt tttattgtaa ttagtcattg catcatggtt15241attaaagccc cttctcagga tgattgttaa aatatacaat aggtgtggta cagtcaccaa15301ccaatctgaa tggatcctgg ctgtacacac ctggttcagc aagcatgctt actggctgct15361gatgaaatag aaccagccat actagatgga cagtatgctc cctcagatca gtatatgtac15421tgaattgatc tctgtaggat ccaaagtttt aatatagtgt tttgcatata atagatacac15481tcatatatag cactaatcaa ctttggaaat ataagaaagg tagcctaagt attgggaaaa15541ataagacagc aaggtcacat atttgtggag tgaattattg gatgctataa taatagctgt15601tattatactc taattgagga tcttccataa tggtttcata ttttctcttt gagagtagag15661cctagcaaat ttctagaatg tttgagattt atttgttctg gaaactgaat ttccattctt15721actgaaacta aattggattt tataaagata cttaagaaaa taaatgattt attaggggta15781tttgttcttc aaaggtatac agccaagaaa attaagcttc atggaaagtt gtgattgtaa15841tactttttaa agatgataaa cacctggttt aggagactta atggatcaag gcagctacta15901catggatgct cactggcaaa aaataactat taaatacatc catcttaact aatggaaggg15961tcagctagct taaaggatgt aattagatgt ttttagtgta aattgggaaa acactcatct16021gtacaatttg aaagtctaat tggtttgatt taaatgttaa agaaaagtgg tcgtgtatag16081ctctcattgt caacaagtgc acaatatttt atcaatcctg aataatgatc atttctttgg16141atgggatcaa aatgactctc acatatactt ctaatttaat aatagctaca acttaaagga16201tgttaactgt atgccaaata caacacttat ttttgcttta catatgcact taattaaatt16261gtggaaattc aatgattgat tagtggagat agtctgaaaa agtattactg ttttatacta16321caacttaggt tgaatcccag aacccttcat agggctcaca ttctgaccca ttgaactaat16381ctcatgctgg gattaaaatt cataaacaac ttctgtgaat aaggtaaaac attattagag16441aaaatgacta agtggaaaat tattgttttt aaattagaag cagaataaat ggacttgctt16501aataagaata acaagaacat ggctatttaa cattaaaaat actcaatgac attctcaaca16561gaaattgtaa ctccctaaaa cattactttt tacattctga accccataaa gcaaaaacaa16621gaataaaaat tgtatgtgaa caaaccgtct aaaagtgatt acttaaaatc tatgatatac16681tctggattaa agtatgctgt cctaaagcaa atctttgtag atgagtattt cttagagtgg16741actaaggtat tcataaaaca aagtgcttaa atgaataata taagtaatcg agatgaagtg16801gaacatttca aggatctata atttcaagaa aactgtgctt atctacagag tcacagtttt16861tctgatgtcc tggtgaagga ttgaatggag gaaaaataca ataaaagaac tatatatttt16921gtctaaaaaa aattgaattg actcgtttgt ttttgttccc ccgctaaatt ggctgtttat16981gttcactgac aacaaaggca aataaagata attatttgga tcactcaaaa caaacaaaac17041actgattaaa catatatctc aagtacaaca caaagttgat tccatatttc tacgtggaca17101aaacaaaatc ttgcactttg cattgatgtg tcctcctatc tgcatagagc aaatacattc17161ttaatcattt tctcctcact ctgtttatat attctttcta atttaaaatt gcctctgaaa17221gtctaatatt attgcagttt tcttctccat attattaact ttttaaaata ttttgcttat17281agtgattcat gcccaattca tatagataYg tagattataa atttgattat tgtcaaatgt17341taatagcaac tcagtgaaaa acaacttgtc caaaatgttg ctaccaaaag ggcacttgac17401tataaataac tcacattata ttacatattc tgtcttgttc ccatcYtcaa cccccgcccc17461tctgctcaat aactaatcta agatcaactg ttgatcaggg tcagttcgct aaaggggaaa17521catattgact cataagttta aataaaaata aagtcgcctc tgattggctc cttcttggaa17581agcaagcaag ttttgtgtgg gactgatatg aaagaagaaa gcagtattag gatgaataat17641ttctgttcat tgacagctgt taacatttat actaaaacaa ttttatttat aaactttagg17701attaaagaaa ggtaaaatta agtagctaaa actatgcatg aattgcttca ctatatatct17761aaattctaga ctaatacgaa aatcaatagc tgctaattgt atagatgatc atatttgtta17821ttcatttgct caacaaacat tctctaagca actctgtggg caaggctgtc tgctaagtgc17881tgggatacca agaggaataa gcgctatctc tggtttgaga gttgttcagc cttatggagg17941agacaaaaat agaaataggt aatttaaatc aaagaacaat gataaaatat gcaaattctg18001tcaactgtgt tttaagagca tagtaggaaa aggaggcaaa attggggaga gggtgaatta18061agtgcagggc atacgtagct tgatgcatcc atgtaaaata ttgtgctggc cctatactta18121tatgctctca gatgcattct tcggtctctt ccttcctttc ttctcagagt agctgccctg18181tacagttttc cagacctctc cgtcaactgg cttccagttg gttttgatca ggcactggca18241gagattggag aatgggagga agagagaacc aggatattte tccccatcct tgtctgtctc18301aggatatatt tctggcattg gctgaatttc tttcatgttt cttgctccca gaaagcaggc18361ttgtcctggt ttcggctctc gctgtcctct agcttatggg gacaatagct atttttctgc18421tgcttcacta tcccccactt agtttctcag cttctttcac gagtatatca tcaattgcta18481gtcatttgtt tcctctattt caaatattca gagccatttc tagtttcctg gttggacaca18541aatgcctgac atactgagtc taaattccag gagtttgggc agcaggtttg ggactaatca18601acatataaga gagagttcat gagtgtacac agagatacgg taggtaaaaa gattggtaga18661ttgagggtgg aaccctgaag tatatagcaa gagataaagg aaacatagct tttgaaggcg18721tccaaagatt aatttacaat ttcgaataga acaagaaaaa tgtggcattc acttaagccc18781ctgagaaaac ctaggtatca cttagaccca gcatttgtca gatgaggttt ctgaagRtag18841agaaattagg tcacatatct aataaagacc agaattgaaa cctaaacctg tttccctatg18901tagtgctctt gtgataacac acccattatc atggtaatag tgtgacattt ggtgtcattt18961tcacttgttc tgtccttgtt ccatgtgtgc acttcagttt ccttttatat aaagccaata19021tatgttatta gttacctcat aggtaattgt aagaattaaa tgagacagtg catgaaaagt19081acttagaata aactacttag aatagttaaa tgtatagaga gaggttggaa tgacacatga19141aggactatag ggctactaKa ttttatttat tttatttttt gagatggagt ctccctctgt19201cgcccaggct ggagtgcagt ggcgagatct gggcttactg caaactctgc ctcctggatt19261caagcaattc tcctgcctca gcctcctgag tagctgggat tacaggtgcg tgccaccacg19321actgtctaat ttttgtattt ttagtagaga tggggtttcg ccatgttggt ctcgaactcc19381tgacctcgtg atctgcccac ctcggcctcc caaagtgctg ggatttcagg cgtgagccac19441cgtgcccagc cagggatact agattttatc acagtattat gaaaggtaag cttgtcatcg19501taaagcttat agatactcct tttatatgtt tgctgacgtc aaggattttt cacatttggt19561aataatttat tatttgttta ataaaacgaa atttgaactt aaatttggtt 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gaccatcatg20461acatttagaa attttgtttt agaatttttt atttatttct aactcaacaa tgagaagggg20521ctttatattt ttatatgttg tactgatgag ttattccagt ttttttggta aaaagagcaa20581tattgaatta aataatatat aaaaatgata ttttaataaa atatttctta tttcatgtat20641ttttttttca actcctttct gtgttttcca ctgtgttgag caaactggct tttgtagtca20701ggctgcttgg tctctgtttc tggctttgct atttcttgct atgggtaatg taaggcaaag20761tacctggtca agactgtttc tccttttgtg atataaagat aataatatta gctacatttt20821gattatgaaa aaaaacacat gaaaaatgcg tgaaaagggt taacagagtg cctgagaaat20881aataagcatt caataaacat gRcatattaa caggtaggca tgStcgtagt agccttagaa20941ttcaaccaaa atctgctaat cacttttagt tttcagcttc aaactttttg gaaagtcaac21001tagatgaaga attcaggata ctttgaggac tgtaagctta gaatgttaat aatttaacag21061ctaaaSgaca gtagaccact tcttatttct tcggctagag aaaaagctac atggcacatg21121aagaagtttg gaatttgcct ttatatRtgc ttagataagg cgttctaaaa agatggaata21181aaaaattgtt acagaaagat gcttgaacaa aagagcatct tgatactgca ctttgtagac21241aacacttcta gcatagtggg atttactaac aattaattat aactttcata ggaagtattt21301ggtttgctag acctgctgaa aatgtaacca tatttatttc ttaataatcc aattatatct21361tacttgaaaa ctgctctaaa ggtcctctta ttctacttca gagaaggtgc tgaaatgttt21421ccgcacataa taaaagaaca catgtaaatt cagaaattat tttatctcaa aagccattac21481attgttccat tataataatt ctaggttaaa ggtttaagta caaattttta ttaagtgtgt21541atgtgaggga ccaagttcat gcaaattaag caatttccaa catttccatt ttaccatata21601tatgttaaac catggtctgc atggtcaaca tctgcataca tatttaagta acacacaaaa21661ttttagctta tgtaattgct tcatcaagat tcaagacaag tagttaagaa aaaaaaaaga21721actctgctct atctgttagc attagagggg aaaaaaagct taatagcagt aatcttactt21781ttatcagtac gctacactgg tagagtgaac caaaacatac cttgttgctg agccaaactc21841tattgaaagt gcaggtcttt ttagtattgg taatctagta gatcagaata ctaaaaaata21901aatattctta tactatccca gacttttaaa aagtagttac tgtaagtctt agtaattgta21961gtacaagtac attttgataa tttttggggt tgttcttgag tcttagactt tatatgttaa22021tggtgatacc cagccagtct ttgctttcaa ttgaaactga ttttaacata ggtacttagc22081caatatgtgt agtaagtaca ttttgaagat tatacaaaaa actttcatag gattgtgaag22141ctaaattaag tgcaaaggaa atgtaactgt gagtcgtgtg aaggatgtgg tttctctgaa22201ttcccaatga ctacacttta gctcagaaca ttctggctat acattttata atttatattt22261ttaaataatc gctagatatt tattaaacac ttttactagc ccaagctata attttattag22321atacaatggg ggatataaaa gaactgccaa atactacttc ttacacatat cctataatgg22381ttttcaaaag taaatattcg ttgtgctcat atggcaaagc aagagtctcc tagacagttg22441tgagaagctt tgatttacaa gagaaacagg aaaatgggaa aaatcaaaat tagaatgtgg22501tatatatata aaatacataa tctacttata ctgcagacat tcaaatattg acaaaatgac22561atatttactt tttggaatta taactttacc atcagtgtta aattatatat tttgaaaggt22621gcagaaaaat tggaaggata acaaattatg tgtggtttaa aacctttcta aattcacatg22681acatttacag cttagaaaga aatttaaaga atttgtagaa attcttttct gtcaaattgg22741atcatcagaa gtcaaagacc aatgggatta gatttctttt aggaagaatt tccaataaaa22801tataactgat taagtacaaa taactctatt gaatccatag actaaatctc ataaaagaaa22861agaaagctaa gatggacagg tgatacaata agaatttgct taatttaaat tgaaaaaaat22921gtagttcaaa acaacaaaat gaactcgttt ttcactgtaa aaatctattt 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aacggctatt26341taggaaaatg tgttcctaac ttatagacat ctgatataca attgactaat tgaactgaat26401gctgttaaac aaccagtcat caaggaattg ctagtaaccc acaggactgg atttgtggaa26461acagaatagt tatgtttctg aatattttct ttattttatg tagaagaaac ccagatggtg26521tagttagtgt taaaaaaaac ttcttaaaag aataatttta aaaatgtaaa ggtaatgtga26581gtaggatgta aaatgatgga catgtataaa gtagattaaa atgcataacc tacataatga26641gtcagattag tcaattgatt ataaacaaaa gaaagctgtc atactttttg atgggattct26701ataatccaat tcataatttt tagtatgaca ttgaatggaa gacattaatt tcatcaattg26761gaaaatcaca atttactatt ttcaaagata aaccttaatt attagacact aatattcaat26821caatttatat ttattaaatt aaaataattt taattcaaga gcaaactcac aatgaacaaa26881aatagctaca ttataaaaat gatttattat ctatgtttca aaatcgtcca tcctttgttt26941tgtcaacttt ggttttacct gatgcgattt ttagttctat tcctcttctg gaaaagttga27001gaaatgtgta cagaatataa aatagcataa taagaagaaa gaaacaaagc tagaaataaa27061tttaagaaaa gacaggtttt ttttccatct tgccaatctc tttgaataga attctcattt27121tgaaagcctg tgcaagtgtt gtcaatattt taaatgcaaa caaataacat 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tttttcctca28021ttctcccatt cagaagtgac ctctcctatg tctggactct tgtagcacat actctgtcgt28081tattggaagc aaagcctatg ttttctcctt ggtaacttcc ttaaaagctt gtcccggaac28141catgtttgaa caatatttaa tgaataagta ataataataa taatacatta atacttcttt28201cctttaacca atcaaaacat ctgaatgatt tagtagataa tttcactttt taaatcaagc28261tttgtgtttt atacaaatca taaatttctc aacaagaagt ccacttgata tggaagcttt28321ttttccatgt tctttgtcaa agaaacttta gggagtcatt ttttttagct tttaaaatca28381gagataactt tactctgtca tactgtatag aaaaaggtga tcattcaact atatttttta28441accccaatct cccaaaacac tggtttgatt gaaggaaata tttaatgaga atacactaac28501catcagagat tataagctta tgaatttcaa aaagttcata atcacaaata ttacatatat28561atttaaaatg aaagtgacaa aagaaaataa tatgtagagt aacattctgt gggagccagc28621actattacag actctagaac ttgtcctgat cccatgtttg tatgtaggta gttggagagc28681tctgggttat atttaagtgt aatatatctc tattttttat gttattgagg gaaataatat28741tttaaacaca ctaagtataa tcttccataa ttttataatt tcaaaatcaa aaatttagaa28801atattcaact tctgctttat tcttccaact cttttaatta gagtttcaga aaccaaggac28861ctactatgtg tgaatgtatt acacacctgt gtattacata tgaaaaaagt ttatataaac28921attgagaggc ttttcttagt aataagatac atctttgggg ccatttttta tgaattacat28981ttttgtgtct actttctttt tgaaatcYta agtgtattac taagtaccaa gtaaaggaaa29041gagtggatgc tataaatata tgatagaatc agctcactca tagaaagggt cctgtgccct29101aaaagaataa atcttagaac gttgaatcat ttttgtaact caattaattg agggggcatt29161tcaggtaaac aaatgttacc ttctgtgatg caattcaatc tgttttaaat tattctttga29221ccatttcact ttctttggct gttagatggt tcaacatcat tgtgttttat tactccattg29281tggctaacct taattaaatc ggtagtgtct ctttgactgc acatctgtct tcaattagtt29341tgctctgttg gcttttcttc cctctaccac acagtactta aatatgcaaa gctatcactg29401tacttaccac atgatggcag cactttgctc tgagagcacg gattaaattg tagacttcac29461ttttactact tttatatcac atccagttat ctgaaattta aaaggcaagg tctctgccta29521tcctactaaa ttccaaaaaa ttagtgtttg gaatgtaaat aggttgctgt ctatgatgtt29581ccattattgg ttttatatct ttggtccagc acagagctat ttccaaatat tctgaggagt29641aatttattat gtaggtagct tgtactttcc caaattactt ggttaataca gtagtaaata29701tcttcctaga tacagtctct aaaattttaa gattaaccga aataaaaagc aatcaataat29761tttgtttatt ccaatactta atgatggaac ggaaataact acctagcagt tttagaaatc29821aagtatttgg gaatttttac cccctgtagc tagctttaat ttagctagta tttacttcag29881gtacctttct cctctctttc tttgtttctt tctctgtcca tccctctttc ctttccttct29941tccctcctcc tgtctctgtc atctacctgt tgttattgct gttactgttt ttgttttttg30001atatttttac taaagggatt ttgcagtttt atctcatgtt ttaattttct tttgaacgtg30061gacctgaaaa tttcaggttt taagtgtcat tgtgccagca ttatttggaa tgttttccct30121tgaattatgt acaatcacac ttcatgtatg ttttctatta cagttcaggg gttctagttt30181aagaagaggt actgcatctt aaaagtaatt tagaagttta atttttttcc aaatctactt30241cagtataatt attatctatt tgggaaaatc taaagcaaac tggacactag aaaaatgggc30301tagcttcaat agagctattt gtagttttca cacaccacca aatcacaagt tttcacaaat30361acaggtgttg aaactctgat atctctatca gtcatttgta agtcgttagg catggaagtt30421gaaggaagga tgtcattgac aatttctttt ttaaatgaca aattattaga tgactattag30481tcattacaga tatattttat atatatatgt aaacaatata tctattataa aggtatgaca30541agtcatgaga aatggaatgt tgcaggcttt tttcttataa aaataaaaat atttccctta30601agatatgtca tattcacata gtagctcctt ttagagaaga agatgaagac gggttgcctc30661catttgctgt gttccagaaa gatggtaaat ctcgtatatt cacagttttc tctctccatg30721cacattggcc ttatgactga attcttaatg acatcttaat gaacaattta gtttgtttat30781aatgattcat tatctggctt tatagataat aagatcaatg atcacttcag gaaacagaag30841taatcgtata aaacaatgta acaaatgaaa atgaagttag cttaactaat agcacggtat30901agaggaattt tttcttacat gaaaaaatac atgttggctc cgtatactat aatccttgct30961tttaaaagga aatcataatg ttgcctttta agtaggtctt cctttttttg aggggttttt31021agttcagaag tgattttgga agaaaccact taaaatatag tttaagtgag aattaaaatt31081gtatagtaaa taatgaaaga aagtttgact aagacattat catagatgaa aattaagaaa31141gtaaattcat tttgtcttta ataaattcag gccgtaattc cagctatgtt cgaataaata31201gaatgaaaaa agaatgtatc cagtgctgag ttccggtttg gtgctataaa atgcaaatac31261aataatgatc tttatcttta aaagattaat agcatggggg atcatatgtg aaagaaaaaa31321cattcaattt tgagctacat caaattttca tgggcgattt ttaatttttc aatcttcctt31381ttattctctt ttgaagcaga agcattttct ttcttgctgt ttgcttgcac tacacttgca31441ttaggctgtc agataggtat ttccaactga caattggaga aaaaaaaaac tcaagcagga31501ataaagacta ctgtaaaaca tgcaggtttg tttgagtaaa aagtcattct aaaaatagtc31561attttgaaaa aagttcttac ataatcctga aactgtgaga acttacatca cgactgctta31621gataacaaga atgctgtcct taaatggaat ccgaacattt tgaaaattca ttaaattttg31681cccaaatgaa aaaggttgag cacatttcca gtttgttttc atattattcc aatttggtca31741atggccacga agtaaagagg aagtttggac agttatttga taaatgcacg tctggtctca31801tgagacctaa cctctctagc taatgtcatt aatctttctc caaacagatt atattaatga31861tctattaatc tataacaatc atttcatcaa tctttctcca aatagatcct taaaaaatac31921tttctgtcta tatatagatg gttatattgc agtggattgt acaggttatg atagaaccag31981aggcagaaca aataaattat aattaattat attttccaga tatttttaaa catgatgatg32041tttggttgca cctaaatgta aatcagtgac caattgcatt tttctttttc ctctcagttg32101aagatagaca aaactcactt ttctgactct tcataatttt ttccagatac ctttaaattg32161attattcaat aaatattact agcatgagag attcatgaag acagtaatat ttgtctattt32221tgtttgctgt agtatcacca acaccttgaa gaggttctgt gacattattt gcactaaaaa32281taatatcagc aaatgtatgt aaaaatgatg acctaataaa agtaattcag atcatatttt32341tcataaatga taaacacgtt tgcaaatatc cttttggcat taaatatttt gaaactcatc32401aaattatata cttgaaattg aggatcaagt atttcgtaag aagctttcca tttttcctca32461aatttgaatg aaagtgataa tcttcagttt ttctattgtt atgccaatac aattaatttc32521cactatatgg gatcagtaat gtagataatg gaagatgaaa ttaattgata atttataaac32581aactgttttt ctataaatgt tactttcaag ataagaacaa aaaattttat tctaccttta32641taacacttct gtcactctac ttcagcttgg aagcagcttg attaaagctg ataggcaact32701tatgaaattt caaaaggaaa aagtaacaat tacggatgtg taaatataat ttaatctata32761tcattcccaa taaaataaaa aattgctata taaattgaaa aaatgtcatt tgtcaatctt32821aataattata aattaacatc aaatgagaaa ggaaagaagg ggttctgaaa tgcaatagaa32881gagatatata agatacagaa aatttggaga caaacatgat aaacatgttt tggaagaagt32941ccttggttat tcagtctttg actattgtaa aaattatgat tgtgcataag tctaagttgg33001tgacttttat atatttttaa gcttaaatat atttaaacac ttcaacataa tttaatttat33061aataatttat aaattataat ttaatatact taaatgttta agctggacaa caagcttaaa33121tattttagtc ttatcttaga acttatttaa taatccaaag aactttatgt tatgtaggtt33181atatctactg atatttatca tattcaaaat aaataaaaca ttaaaacaaa aaatcaacaa33241gcacatattc cattagtcat caaagcaatg ataacatgac accttatgta gctgctggaa33301aactccactg tacactcgag aaaaaaatga gagcaaaaaa gcaaataaca tgttaatctt33361ctYaagaata ttttgacctt gcagatccct gaaagggttt ctgggacatc ccccgcccct33421accaggaatc agtgagaatt actggtcaag acccacagtc atggaaatga agtactgtga33481aaaatgagaa gaaactgatt agcatcagtt tctttgacag tcaccttaac aattacatat33541gtcaagatca ttttatcatg ctatttattt ttaattttgt gttttgtact gaaaaaaata33601tcaagttttc agctttcttg aaRagattca ttattgatat tagcattcat tttaattgtc33661tctaagctNc tNgtttcact cagaacttca caactctaga aatatatggc atgaattaaa33721aatatatttt atcataaaat cttgtctatt tttggttaaa aacgaacaaa acaagaaaat33781aactatgtta ttgtgcacag aaacccggag taaataatga taattggcag gcataaaata33841atgcagattc ttccataaag aggttttgtg ttcataaagt atcttttgta cattaataat33901agttggatgg aagcaaaatg gtgattcaat agttattgac tttattaaat gtcattggca33961atgaggatgc tttaaaacaa aaactacaYa atagacctat atagaaagac atttgaatta34021ggtagttgta aaaatatgtt ttctatattt ctctatattc caagttcttt ttataaaaag34081aatcagataa aaatacagtg tgatctatag aacttgaaat acatggtttt caatttgaaa34141tgttttaatg aggaaaagtg aattttgcta tgttcttttt aagaacaata tggtagaaat34201tatatttctc tacctttctt atacatttgg aaatagaaac ctgtgatcct gacttttacc34261aaacatttct tctgaaaaat ttttctgcct tgtttgagtt gtggtattag gttggtgcaa34321aagtgattgc cggttttgcc attactttca atggtaaaaa ccaccattac tttttgcacc34381aacctaatat tattttgatt aatctgttca aattataatt aaattaacga attatacttt34441tcagtatcta aaacacatta aaatataaaa tgatggttag atctacgtca ttcctattca34501cacatctacc aaactcaaca ccccgatcaa aattttatat ttcggatata atttggtttc34561atttgaaaat tcatgtgttt ttggcttttt agtgaYtcat tatatattta catgtacagc34621acagtaataa aaatggaaaa gaatattttc atggtcttgt tcagtataca tgaaaattcc34681cttcatctta ccccataccc acgtttcttc ccccagatca tgttgcacaa tgRagaatgg34741gtgaaatatt atggtttaca tgatgtatgt aatacatgtg gatgaaaagt ggaaaaataa34801ctaagcattt catggatagt tacatatctg gatttgggga ctattattaa atcaacaatt34861agtgtctttg gcattcctct accactcctt gggcagcata tccctggttc catgatcctc34921ctatccaagt gagaaagttg cctgccccat gctcttattg taccctgact gccccccact34981gtgtctccat tggcattctc attgccctcc cccatcagca tttacttgcc tgtcttactt35041ccacaggtcc tgctccctgg tggtaatttg atgaactaac tttttgaaaa taattgcatt35101acaaaaacta ggctttatgg gtattatttt taaagacatg tcataaataa tatttaagaa35161accttggtat gttaaaattt aggcctttct cttcttctta gatcatatat taatagtctc35221atcctcatta tttaggccca tagctagacc atatgactaa ggcataacaa ttttgaagcc35281aagtttggtg tttagatggg ccagttggtc ttacttttca cgtcagatga aatgaacata35341taaatcccca ttgtggacca gaccaaagta tgcagatggc tcggtgcaga tccatggcta35401ctactggaaa agaacagaag tacccattga tggtaggtca gtgaattcat cttcaggaat35461gaagatccat aaacaagaga tagtgtgata tggtcagagt tctacattta ctgtctgatt35521ctgtatgtat agcagaggta tttccagcaa taaatttaaa tgactataga aatatttgaa35581aatgcatagt caatcattcc aagaaaatgt agaatctctc aaaaccaaaa cgttagaaaa35641gcattttgat tttcacagac aacaaaataa atccttatca ccagagaaat aggtatagaa35701ctctgaaagc tgaaattgtc caaattgatt ctctcagatt ttgttggatg tttgctatgc35761gcatgtcatt acagtttgaa ttttcttttt attttaaact gctcaaggtt taaaaaattt35821ttttgcacaa acacatggaa atgaggaatg taaaatttgt ttttattaat cttgtacata35881caatgttaaa gaagacttca cctccaaatg aaattgattt ctcttatgct ttttccttta35941tttttcttat gggcctgacc tttacagaaa cagggaaggc taaattcctt ctgaccagaa36001gtgatcatct gcttctggtc aaaagaaaat gggatgctga ttagctacta tcatctgtac36061ccagcaattc cctccactcc cagtcccacc cactcccaaa cccatccacc cactaccact36121tgcccaaggc agcagctccc acagagtgat ggcttagtaa gagttcacca aggcaacaca36181tggatatgaa actcttcttt gacaggtgcc gcaaatgagc tttattgaaa gcggagtttc36241cagtagtggt ctgtggaagt agcatcaggc ttttctccta caattcagcc acatcaggga36301actcgaaatc atatttcctt gcatgaccta tttaataatt tattgtctca cctaatctct36361ccaacctgtc ccagtggctc ttgttttatt gaccagggta gagggatatt tctcctcatc36421ctgtattgcc tatttctaaa tttaaaactc atacctaaaa tcttagtatc aaacaaaaat36481agctcaggtt tttcaMactt aaatttagta tattggattt caactaaata aatcttcaag36541gttttaggaa tgcttcaact tgttttggtc ttgatatagt tttcaaacaa accaacaaat36601aaaaaatctt ttgaaatggc aaggtttgtg ttcaaccaaa ccaactagca tggggaaaac36661aacaacaaNa accgaaaaca tttccagagg tttttaacta cggagagttg tttaggcctc36721aggatgtttt tgcacaatct gtctaatctt tcccgtcagg tgttttagca tacaactaaa36781ccccagaggg aaagaaaaaa ttccttttga ctcactccat aattcttgtt tgacccatat36841tttatgctag ttcaagcaga aatctgctga gaagcttaca aaaaatcttt gaaaagctct36901ttagggacaa ttttgtcact cttgatagaa atgcctacag atggacccgc agccttttct36961aatggagcta caaacattgc ctaatgcctc actgggcttg cacagtggga aggcttcctt37021ttgttgggtt aaatcaacaa cctttagaaa ggaggaccat gtaggcagag agtataaatg37081gaattccgcc atcatttcca gctacctcta atcgtttgta agaacttaag agataagtca37141taaataagta catgactgtt cttattaaaa ttaggagagt ttgattttaa aggagacaat37201gtattcaaaa attataactc aaaaagtcac aagacaatgt tatatttgtc ggttctgaaa37261atttctagct gtgtgttctt atatcagcca gcctacccct ctgactctga ttgctcacgc37321ataaaacagg gataagaata ttgaaataac Yacctcatcc acttaatgtg ataaaatgag37381aaatggatga caaaacactc tgtacaacat gaagaactat tcaaatgcag agtattattt37441tcagaatgcg gatgggatta tacgaactcc attgtgaggt ttgaaatggg acacttttat37501ctcaacactt atgattatta ctaatacttc tgcagttagt gaacaggaac ttgcaggttc37561tttttatatg tttgtatctc tgcctagttt ttctctctgc actttaaaca aattatattg37621cattttatat aaatttaaca ctatagtatg tttgcYtgaa gtgcctggaa atatacatgg37681atcaattata ttgttacact tataatagca gaaaaattat actttagaaa acaaggcttt37741ttaatttatc gtctattaga acaaaattaa acatgaaagc atgctgtgtt atctagtttt37801cctttcaatt atgattttaa tgaagagtga cttctaagtc aatcacaaga ctaaatgttt37861gagaatatct attagaacac tattcaaaaa aagggattaa aaacacttta atataaattt37921taaggactat gtctaggaag aaaatcatat aaaatgtcat atagaaacac ttaagtcaca37981ataattaaaa ataaggtgtg aaataaggga agtgtttgat tactgtgaaa tttataattg38041aaagaatatt taaaNaacgt gaataacatg aagaaatcaa ctgaacttat ttgatggaag38101aaaaggtgag aggtggaatt ttaaaaatga aagttttagt aaggctaaca agaaaaaaga38161aattgtgctt tgatgataat ctataaactg ggctgagtaa cacatattca gaattatcct38221aatgtttaag ctgattgttt ccttaattac ctaatttagg aaacacctca gtggtttacc38281tctgtaatgc gacatatatc ttacattgcc accacatata taattaatag ggagaaggac38341atatgaaatg tttttaattt tatgagaata aaaagaacgc aataggaaag tataaggctt38401gattttaatt ttgtgccaaa tatagactca taaaatcaaa tattttaggg ctgagaggta38461cttaaggggg caaatagtac tatcctattt taaaaaaaca ttattttaat atgtgctttt38521gaattaaatt tgaagataat tgtataatca ccctagcttc attccttgtc actggcagtg38581gcactctttc ttgggggaaa attctagcaa tgttggggtc aggttacaga tggatgtaaa38641tttgctttcc tttctgaaca gagctatgag ttgaaggttg gccacttctg caaagcaaga38701gctcatggaa tttaccttcc tctatccagg aacctgagaa gattgcaaag tagatcagac38761gtgggagaga gaaatagcag aagtgttgcc tagggcagcc caatcccatt atttatttac38821attttaatgc aatctctgat gctttcattt ggtgacaatt ttccatctat cttatagtgt38881ttccaagatg gaactttgtc ctgctctcaa atgtccaatc attacagctg tcactagcaa38941tgaggcagta attaaaatct attgcaacga aatcaattga gaaaacaaaa ctgacacatt39001cattttaagc tgcatcatat aacaatggct cagaacccag tagtttatga atgaaataaa39061gatactttat ttattacaca ggcacttgga gattcacatg cccaacacat ctgatacact39121taacaactat ggaaacacga gtggaaaatg tctgtttgta ggatcaagag agaactttca39181ccatgtaaag gtaaatgagt gcattaacaa tttttccttt actttcctct aagaattttc39241aaaatccact aaatttttaa aagttacact tgaaataata ggcaaMaaac actgatccac39301attccaactg gtaggctaac ttgaactgtg agcaatttaa acttttatca caatgaaaca39361tatttatagt catgcttgaa cagattaggt tagtagcatg cattccattc tgcaatatga39421atattgttaa tctatttccc aacaaaagtc aaaagatgcc cagtgcatag aaaaatacaa39481atcactgtag tcaatatatt acgggactga tcttgggact tacatggtga tgatgaagtc39541ctgaaatgcc atgcccacaa aagcacatta aaaagtggat aaacatattt tatcagacaa39601tctctaaaaa ttaaRgatga gagctttaaa tttggggatg ttcttgctgt gtgctttgaa39661cagggccttt ctgctcacct ccttctggct tgactggtcc atcagacatc accttcctga39721gggctagctg gctttggtct tggttgccat tggctggtgg aggcagatta tcagactgag39781ttctttgaat ggggaggact cctgctatgc tgtgtctgtg ctgcttcctg gcatgattag39841actgaccgct tttatgtgct gctgtggcag tgtgggtgaa atggaaaccc agagtatgta39901tcctcagatg gaaccagctt aaatgttatt catgcaaaca gaagtgctct tttgctctat39961ttaaataggt tgtgttttca aatgccaaaa tgacaccaca tgcacagcac taaaggggtt40021taaaactggc tgtttctttg aaaatgatga ctagggagat ggagagaaca atgatctatc40081tccattgcct ggctaatgcc ttttagaagt aacagggtac ctacatttgt gaaatatttg40141cacatattaa gtttttatat aattatatca atcgactaaa ttttcacaaa tgaaatgaaa40201atacagcagt taagataggt caattgttat actttgattg acttatagct accagacata40261gtatatcttg atttccaatg gttagaaaaa atatctgtaa aatctgtcat cacatacaaa40321gtgtgatcca acagaatgga cccaattgtg ctcctcagtg aggttgcatt gccttatcac40381atatgcttat acatatatta atttaatgta gttttttgag aatgtaagaa ataagatttt40441cttcacagat ttttcagaag actacaattt tttgcacatg gaaagctacc atatttcaga40501agcaaaattt gttagtgttt taaagcactg tcttgaaatg gcttttcctc caaggttcta40561gaagttgact gttggttgca ttatcttctg cctagacttt ttgagtgttt tcagatacaa40621gaatctaatt gcaaatctgc tctaaatact tatatgtgtg cacttttttt tttttggtat40681aagtcctgga aaactggcag ccacttgtgc ttataatttc tgtattctaa aagatattta40741agcgcaaaca aaaaggacac acaacattgc tttgcttcca aacatcacag acactgtatc40801aaatgctaKg agccaggttg gatggatggt ggaaacaaag gagaaacagc cagttttatt40861tcagtctatg taaataaaaa aaatctaaat aagaaagagc atattaatgt atttatatcg40921ttcttacaca gacaaaggga cagagaacag ggtgagcaga ggatgttgaa aactgaaagc40981aaacaggaaa ggacagcagg aaagaaaggc aacagatgaa cattaagctg ccatgctgag41041gaatttattt gcgtctttta cttggttgaa tgcggagttg ttgcacggca gcttcggcag41101cagctatggc agcccctgca tcttccaggt gggtctgagt gacgctggtt ttgctttgac41161tgcgagatgg tccatgagaa cggagatggc cttctgatga tgattttgag ctaccaggac41221tactatggga tttctcaatg gtgggaacct gcatgtctgg tcacaaaagg gtaaaacatg41281agttaaagtt gttccagcat tcctcctatc aagcttggtt tacacataat ttgaaactgc41341ctaaaattta tgttaattct gagtcttata tgattcattt atgcaactgg tcatcataca41401gaatgatgat taaacatctg ggaaacaatt tgggggagga aaggcaaaag gatgatagca41461atgagaagag agaactttga aaaaagaaat acattctgac tagcttctag taataaatat41521gttaactttt agtttcacca agcacctctg ctaattccag cagaggtcag gtaggagaca41581atacacagga ggagttttaa gcaaaaacag gatgaaggat gtgaatcact aggacatatt41641aagtgtggag attgtagaat catgcctcaa accattgtag agaatgtatg gatcacgtat41701ttcctaaggc agtcatagaa agagatcttt atactaatga tcatgcagta tgttaagcaa41761gataagagtt gagatggcta agtggaaccc ataaatgatc tataaaactc attcataaat41821aatctacaaa actcgtagtc agcagtttat gatatcaact cagtggccta ctggacatgg41881actcactttc tcatctccta gaacacttgc atttatgatt cagagaatct catgtacaaa41941gaaccacttt acaatttgct ggactttcct agaaaaggtg acaaggaagt aatactcaag42001tccaatataa aaattatatt taatacttac aaatagttca gtatatctag gtaaggtacc42061acWtgtttta tcccacagag gcagtagctc agtttatgat gaggattaat gttgtgttgt42121aggacagttg gttatgaacc tctcaaagga aaagctcatt tgaatgagag tgctgtacat42181atttgcatat tgccaaaccc aatgtatggc actgttattt aaaaaataat aatagttatt42241aaaaaattag cttgagtcca acatttagga aagacttaag taaatttcag cataccaatt42301gttttgatga caggggaaaa tattctttgt atgataaata aaaagtagca tgcaaaatta42361tatttattgg gtaagacaat atttatggac aaccagagat tcaaacaatt agaaggatta42421taacttaatg ataatgatga ttatgtctag tcaatgaact cgtagggatt tttgtattat42481ctttatgaaa tacccaaatg cattttctga aatcatgtta tcatggattt tgaggtggaa42541tagaccaaat agctcatttt acatgttgaa attggacagt tagagaagtg aagattacaa42601agtactacac ctaagtatgg gtgatgatct gagaataaag cccatttctc ctgattccac42661tctttgcatg acattatatt aagtattttc cagtcgtgtc taatctatta atataggtta42721aatgtacaaa cattcacact aagaagatac agactgaaca aatagacaca aaactttttc42781tacttaaaaa aatatttcct accctttgat gggtcaggga agataccatg gcttctgctt42841ttgataacag atggctttgg ggactgctgg ctgctctgac tggaatgaga cttgccatga42901tcaatgcttt cagtctgttc tttgagagga taccaccttg gagtgttatc gaggtgagMt42961gtgctagata aatcaatcaa tacctgaaaa aaagtgtaac aaataaatga aatttaggga43021ctaaagtata attgattcat tactttttct atgttctaaa atttattcaa ggatataaac43081caaggaatac agtataactt ttgaggatga tgattctttc ttcctcagtt ttcttcacca43141atgatactgg aaacaatgtc ttttacatac tagatgctta ataaatgttt gcttgaactg43201aactgtataa aatttttgac tattaactaa attatttgga tatgatgtta aaagataggg43261tcacaagtca atctgttcat aaaacaaaac ttctttttgt tgtaagaact tattttaatc43321ttacacggtg aaatcccgtc tctgctaaaa atacaaaaaa aaaaaattag ctgggcgtgg43381tggcgggcac ctgtagtccc agctgtgtgg gaggctgagg caggagaatg gcatgaaccc43441aggaggcaga gcttgcagtg agctgagatc acaccactgc actccagcct gggcgacaga43501gcaagattct gtctcaaaaa acaaacaaat aaacaaacaa acaaaaactc ataacttatt43561ttaatcttaa actggactag caaccttgaa aaatgtttga ttttagtcac aaaagggaca43621aaatggaaaa gaatgagtaa gttgcaatta caaaSttatt tattctattt taaaacatac43681tttagttcac atttgaaata gggatatgac taccacattt tgtcatggtt taattggcaa43741cttttttttt cttagtacat tggtgcattt tacagttgct gacatcttag atatgatgag43801ctaaaaagaa atgaaaacca gaccaatgga tatatgcttt aggctcatgt aaacataaga43861ccataaatta gaaggacttc agtaagtcac ttttttgaaa tgaaacataa agtcaggcat43921aaYactaatc tagagtactc ctgcctagga tcaagattag tccaagtggc tagagctgga43981tggaactgga aggtaagggt caaatgctcc agttttggtg aaaagagatg tgcaaagtct44041taagaaccaa acaagtcctc attatcacct tgattacaca aaaggatgtc gtattttgta44101ggttttggat tttttggaga ttatttttaa cattaattca taacttctgc cctaagcaga44161acactaaaca atgtgtatgt gtgtatatat attatatata ttatacataa tatgtatagt44221atatatacat tgtatatata acgtgtatat atacattata tatacactgt agacatacaa44281ttgtatacac tgtatagtat atattataca ttgtatatag acatatatac attgtatata44341cacatgatat atacaatata taaaatatat gtatatacaa tgtataatat acgtatatat44401aacatataat gtaaatgtgt atatatatac acaatgtgta catatctaca ttatacatat44461aatattccta tatataaaca atatgtgtat atattatgtt catatatata aacaatgtat44521atatataata gattaaaaat cacaagatct gttcaYgcta ccaatttcat gattatttgt44581ttatttttaa atttttacaa gtacatagta agtgtatata tttatggggt atatgagata44641ttctgataca ggcatacaat taggttattt tgattccttg atgttaatgt aaaaattgtt44701tttattgttt tctcctactt gtagttttgc ttcttaaaat gttttcagtg atttttctca44761gaatacttaa ttataaataa caatgcaatc aatatgtcac atcaaatcaa ttaaagaacc44821tgtaaatgca tttttaaaag tccaggccat tttataatat aataaatttg ttacttcaaa44881tctcatttgg actcacagta tgattacata tcttcctatt gaaagaaaat ttcatcttag44941aaattaatta taaagtacat caagtgatag agtgaaaacc atttcttgaa ttggtattaa45001tctactcaaa tagtaaatca ggaaataaat tctatctgcc agaagatttt tttcagaaac45061ggctatctta taaacttcag aaacatgctt tgcatcgtgc tttaattggt ggtttaccat45121agcagcaaat aagggaaagg aagtcRgagg cttacctccc caaggaagtc gttggatgaa45181aatctatcat aatcccaaac tgtcacctcc agtgttttct tcttgagcta tatagttcaa45241atagaaatct aattaaacct atctttccat catacatttt cattacacac atacagtaga45301catatttatt tttttcctta aactccttcc atttatgtga aaatacttaa aattttaccg45361aaaactatgt agtttaaata ttaaatatta tgtaaaattt gaatgatttt gcactctaaa45421ctagtgtttg atcatgggga acattattat tttgtgaata aaatgataat tcttacttat45481ttattgtatc cccacctcat tttcaaaaat agaaatcaga atttataaat actctaaata45541taagaaattc atagattgca aaaatgtcta gtattatcta gatgaagtat tggattaaat45601gtttcagcaa tttccaaaaa acttagtagc attgccttat ttctgcaggt gagtttattt45661taaattcaca aacccagatt ataatttata tccagcttca tctttgtaat gaaattaatc45721tactagaaga caaattaaga atctcatttt gaaagacagt tcacttgtaa attggtttat45781gtgggaactg tctttatagt tttctctctg aattcaacct tatacatttc agttattgtc45841tgttaagcaa agatataaaa atgcagattt tcagaaatat aataatgtaa tttttgccta45901cagttactta tctStggtga taatccttta tactagtcat cattttgcct tcttctgtag45961agattagcac cacaatgcca gattcttagg aggcctcaat ttagcaaaga gatgagatcc46021taaaatattg tctagtggag taatacagtt ctatgcactg aatcttattt tttttgtgta46081ttgtgtatga aagtcaaaat atagccagtt ggaaaatgtg tttctctctc aaatatgata46141aatgtggctt agtctattaa aaattttaaa ttctgaaata acaaatggaa aatattccac46201tagttttctt attttatttg tttatgtttt ggctgctttg tcaaaatgtg cacatttact46261ttcaccctgt gcttaaatgt cactcatttt tttccattgg ctacatttct tacatgtatt46321ttactcaact ttttttgttt gattggcaaa atgtatacag atcccaattt ttgaataatt46381gtgttatcac tgtacttata ttagcctaat tctgtcacct agaaataatc ttgacatacc46441tgttccatgg aaatactttt ataaattact gtttgattcc actcaggatt aagacttttc46501tggacatgtt tagtccttct cttgtactca gcactgaatt gggagaaaag aaagagttat46561cttgattatt caccttagtt tatgacttca cagttaaact aaaggacatg cagactattc46621aagatatttt atttcaaatc actatataat tatctcatca tcctcatgtc ttaaacctta46681gtttaagttg aatgcacagt gacaatttaa aaatatgttg gatgcttgta gttttctttg46741ccctWatgat ttttttttaa agaaatgctt gtaggtgttc taaatatact ataatttcag46801aaatcttgct cattcctgaa tgtaggagat aattacatga actaaatagt aacatcaatg46861ttactgcccc ataaacaata atttggtgtt taaccttact atagttttgt acatacRgaa46921aagcaaaact aaaacattgt aatgagtgta ctgagaaaaa gatgaagaat ggctgttgct46981gtggctcaca ttccataaac acttgttact gatatctaaa gtatattcca acactcttgg47041taaagtatat ttaatgcagt tataaagcaa ttttaattaa atataaattt ttggtttatg47101tcaatatgYt attttacaat tgatgaacca aataaaaatc tttcctcaat tattagaact47161tgggacatct gcattaatcc ttgactttcc atttcctctg agcaccaatt agaagcttat47221atgctacaca tttaaaaaca atttattttc aaagcctatc ctaatgaagt ctggcagaac47281atctatagct ggcaccaacc tgctgatggg gttgaaatta gattactgtt acttgctatt47341accaaatggt gcccacacag acaaggcctt tctctaactc tcactgcttt tttctgaatt47401ttctgcccaa gcttatgtct cacgttctaa ataatacttc atctgtggtg ggcactagag47461tcctgtaaac caacctctct gtctgggggt cttgacatgt gtctggattt cacaagttgc47521ctttccctaa aagggggctg gaccctgact cttgtaatct gtagttctat tccaaactat47581agccagacta actcttgcta cattcctggt ctcaagtatt gccatatcct ggatgcctag47641aaattttccc cagccactaa tttacctata cattgtagag aaagctattg aggatcaaac47701tccagctcac cactcacttt taaagatggg cctgcaggca agtactttcc ctgtatctct47761gattttattt tcttaccagt cagatacaag ggaaataacg ctcaacattg actgcataat47821gggatcacct gagttctggt ggtgggaggt gggtcagtca accctccacc cacttccctc47881ttgcccctgc ttactccccg aaggaaagtc tgacttgatt agtctgggat gaggcttgag47941tattaggatt tttgaaggct cgcaggtgaa tctaatatgt agcgaagatt aagaagcacc48001gagttggtgt ttcataacac ttcacaagtc agccattgtg agctgaatcc aactgaagaa48061ataaagcagc attcctcgca cttcaaccta tttgaatgta cacaattaag ggcagaattt48121gacagctgtc actcaagata aattacaaag aattttaggg aaatatacag acaatgattt48181aaagaaagag gagctctcat agactatgga taggaagttt ctatatatac tttttagata48241aatttgtcaa tatctagtga agtttaagtt tatataccca aacatccaaa tattcaattt48301ctactaattc cacttctgcc atagagaaat tctcctacct gtgaacaaga aatttgttca48361agaacattaa ttgcaacatt atttcaatag taagccaata gaagaacgaa taaatacatt48421gtgatatatt catttcaatg aaatagtaag tggcactgaa aatcaattaa attttaaaag48481cctccatatg tcaagataga tacatcttga aaaaagtttt gatcaaaaac ttaatatggt48541atggtataat ggtataaaca taaaatcatg tgaaataagg ttacacatta tatatacatg48601tatatatgac atacatatat attaatatcg taagaaaact tgtaaattat gaatattaaa48661ttcaatttag tgagtttttt aatggatagg gaagtgattt cagagggctt cagggtatta48721tcttttaaaa aatctaaaac aaatatagca aaatcttaaa atttgacaaa attggatagc48781aagcacacag gtatttgtta gggtattttc tataattttt agtaaacttg aaatatttta48841taatgaaaaa ttcaaaaatt tttagtaaga tatgttcctt taaatttagt tttggattct48901ttttttgatg gtgacttaga actaatgact taatttgatg gtgatagaat tagtgtctac48961aaaaaagtgt ctacatagaa ataatacata taaattaata catagaaaga gttaataact49021ttaatttgat agtgacatag aaagaattag tgacgacttt taagttaata gtaaacttgg49081attgctactt tagaatcatg gctattctca taggtgaata tacataggaa acaaacacat49141ggtagtatga gttttaaagc ctctgttttg gacttactta tcaaggtggt gaggttatgg49201aaaccatatt aagatttata atacactgaa cagtaataaa attatgtttt ttaagttcta49261aaaaatacat aatttgttta acaagatggg aattaatatc aaatgcaaac ccatgtacta49321tcactaaaat tctgggtctg ccaatacata ttaaaatagt ctgaagtagt attaaaatac49381ctttgaaggg atttatcagt cactatcttt tcttaatcca tacatgtgaa taattgttat49441tcaaattttt caaatccaca tttagtcttt ttttcactta tacacctagc aaaaattagt49501gaatgccaac tctgtatcag gaattgtaga tgtctatcaa ctaaaaaacc cttaattgta49561ttgacatatg agtatatatt taattcatgc atgaaaagca tttttaaaag taagcatatt49621aaaatcataa ctaacattta ttaaatgcct cctttttcct ctggtaaagt tctaagtgct49681tttcatgtgt taactcatgt aatactcata ttaaccccat tatacaggta ttacttctta49741ctccatttta taggtgagaa aacttgaagc tcagaaaaat taaggYgtat tgtcaaagtt49801gcatagttat taactggacc aggatccaaa cataggtaat ttgatttctt gcttttaaat49861aaatgttata tgaactcaat tgattaggcc catcataaaa tgtgacatcc atctattttc49921ttaattgcag gcttcactac ctgaagtatc tagtttgtct tctttataat ctaaaattct49981ccatctcaat ttaaatacat aatttttata catataatat aaattatgta taatgtgtgt50041agacacgcat tataaacatc gtgaacaagt taggccttct ttagagttgc ctagggaaat50101tctgaatagt acatccKaag tgagtgttgt ggaacaatta cgcctactta ttgtaaatac50161tgatgtttgg ccagatacat ttagaaataa cctaaaYtga ttggacaaaa gttaaaatgt50221tcattttaat agagtatctt aatcatagta atgagtttga ttctccaatt ctctccagac50281ctaaaacatc ctttagaatt aatgaaagta aaaacagctt gctcctaaac accaatgaaa50341gcctttgcag tattttttgc ccttgcattt cctgacatgc agagctggtc acacaaagcc50401attcttcaat aacagttgtt ttataactac catgctctga tcatatcata cacaatttca50461atgaactgtg ttgccaccta acacaattta aaactctaca caaaatagtc atatcaaaat50521ctgtggagtt gaaggtaggc aggKaaacag ggggattctt aaatgtgagc ataaagtcag50581gctgagaaat aagtggagtt catcctccct ttcctttctt gtctagtaaa cacactatat50641tctcttgtgg gaagattaca ttaatgattt aacattagga caattaggac aattatatct50701aataattaaa gaagtttatt aataagggaa aataaaaagg tcaatacata aagatgggac50761atggtgctta tcaaaaaata tctaaaactt caccctcttg aagttttgcc ttgtttgttg50821gtttgattat ttattttttg agatggagtc ttgctctgtt gctcaggctg gagtgcagtg50881caatctcggc tcactacaag ctctgcctcc tgggttcacg ccattctcct gcctcagcct50941cctgagtagc tgggactaca gacacccacc accatgcccg gctaattttt tttgtatttt51001tagtagagac ggggtttcac catgttagcc cggatggtct tgatctcctg accccatgaa51061ccaccctcct tgggctccca aagtgctggt ggtttatttt tttatttgtt tttattggtc51121attttgtcat gcataggtat atatactttt aaggctgtct gaaaaaattt ttgaaattca51181attattctac tcaagaccag caggaaaaca gtttccatgt acatattttt taatttcaaa51241agaaaaactg agaaaagata atgaaatttg aaggcttctt tggtgtatct actttcataa51301aattRtctaa aacattgaaa gtattcattt ttaatgattt atctggtttt cctttactaa51361acttactaca cacacacaca cacacacaca cacacacaca cacacacaca cgtttttcaa51421aattactcct ctgaggttat ttctcttaat ttagaaaaaa tacaacttgc atcccatttt51481gtaagctctg tttaatagtg cgatggtgtg cttattgtct atcattttat tttcctcttt51541gctctctgat ggcagagact atatggctac attgtcaatt gtaacccctg cacctggcac51601agtgtctgac acatagtctg tgctaagtaa atattcagtg atccttagaa catgcttgta51661ttttgttttc ctttaaaatc tactgaagta aaataaattt tggacaccat attgtaaata51721cacacaccac ctgtacctgc tttccacagg tcccagagga aatagtttga taggatacca51781atgttgttta gagaaaggtc cagccaggaa ttgggcttcc aggtataaag atagctaaaa51841ataattatat tcataacaaa atctcagtaa tgagaaaata ttttcacttc gctcaacagt51901gtgatgagag aaaaccagag cgaggccctt catatatgct catttccatt tcttaattat51961ccaacaggaa gtattgagtt cacgcacgac ccagaccttt tcctagtcat tctctcacct52021tatcttgcct gacattttgc ccctcaaagt ttgagtatat tgatcagagg gtgggaagaa52081aaaaRtaaag gagacaagga aaccaaatca gtgtactttt taactagcta acaggactct52141tattttaaat ttaagaatga aagatgtatt acattttaat attaaagaaa agttgtttaa52201ttttctatat agtattttaa aataatttag tataattcag aagctcaagc attctgtacc52261tttctgttgc agagataacc aaccactggt cgcaaaaaaa tgtatgatac acattgctag52321caataataat aacaactact attgattgaa agcttattat gttccagaca ttatgcttat52381cacttttgat gtatttattt catgtatttc acagaaggcc tgggaaggga gctgttatta52441ttcctatttt gcagaagata aaatagtttt agagaaatct taaagcttct aagtgacagg52501gctaaaattc aaacgcaagt tcttaaggac ctcacttatt ggcttccaca gtctaactta52561caatgtcagt accttgtcaa tataattccc tcttacattg acaattctca aagacatggt52621gaaagtgatt tctagatgaa agatataaga aagtagcttc tagatctttt aaatagttat52681ttaagacatg gtattattta taatttataa aatattc~ct cttttataag ttaaatacaa52741tctctatcat tctgataatt ttccccaagt ttctatatat aaaagatagg gagattctga52801ataatgtgat taataatatc tgttctaaaa ataacactat ttctttgaaa caaaactaat52861tttgcatatc agcctgagat acctggattg tgttctgtga tgagagaacc agccataaca52921aggaatctat cacagcgtca gttacattgt ttgccataat tatgtggctg taaggaaaca52981aaaagtttct gggcaatagc aaggctgtgg taacaaatcc tattgtttga aatatttacg53041tatttatgct tttaaaaatc agaaacagcc tcatttggga aacagttcat tcgtagtaaa53101tatgtgaaat tttaatgtca tagacaaaaa cgaatgctgt accattaata atctgggtgg53161gattaaaatt cccatcaccc tattgctaca gatcatacat gatttgcctt cccaactaaa53221tgacacattt aatttataat gcaatgagag atgtgcaaaa taatgctttt ggctatgtac53281taagaaattg tcttagttga cagtgtaatg atgcaacttg tcatgagctg tattagaaaa53341cgctcctact attgggaaca aaggaagatt aacttattgc taatatacta cagcaagcca53401attatggcat cctacattgc atagcagaaa ttttgtgggg cagaagttac aaaggcatga53461aaatgtttca acattaggtt caagtgaggg accaagttca aacacctatt atcttttttt53521ttNtttgttt gtttgtttgt ttgtttgttt tttgaggctg gagtgcaggg gtgcgatctt53581ggctcactgc aagctccacc tcctaggttc acgctatttt cctgcctcag cctcccaagt53641agctgggact ataggtgtcc gccaccatgc ctagctaagt ttttgtgttt ttagtagaga53101cggggtttca ctgtgttagc taggatggtc tcgatctcct gaccttgtga tctgcccacc53761tcggcctccc aaagtcctgg gattacaggc gtgagccacc gtgcctgatg ggatatacac53821ctgcactgga catccagaag acScattaat tacttttgaa aaatggctac agatattttt53881aatagaagac tcgagaactt aaatcatgat tattatgtga gaagctaaat gagttttagt53941gaatagatta tgagacttta gttatgtcta atttcatggt aataatattt ttttcattgt54001ttctatgctt ttaatatttt ctacttttct gtgttctatt gcacacatct gctaaagcta54061tttagaaata tgctcagaac aatttgtcgc agtgctgatt caatagatac aaacatgtag54121aaaaatataa gttacagccc atgttgaagt gccagtgaat gtacataaaa atgccaaaag54181tataattcaa gacatagcag cagagcttga cactgaaaga gaaacaactc taccttgcat54241tctggacaac catgacttga ctgcattgat tccaagagcg agagtgaagg ggtaaaacag54301gaaagtagaa aagacaggaa gaaaatagtt aggaacagaa ataagaaaga aaacatgaaa54361ataataacta gaggagetat cccataacat aaatgacatg aggcagaaaa tgctagaact54421acagtgaaaa taataccttg caagtgtaat acattccaag gcctttgctt gtctaataat54481taggMgtgtg attaaaagaa agacccagga attgaatagg ggtagcttat tagctgcctt54541tgagccaaac ctcaccaagt agggacgttt tatttgacaa attgcatgag gatagcttta54601ggtggggcat ttgttctcca tttcaccatg gttcccacca ttccttgttg cttaccccag54661aatggactga cagattcatg ttacctgcct gtccttcctg gttctttaag tttgtaacat54721tctaattgac acatatcctg taataaataa aagatagata ctcccatatt gtgtaaaaga54781tgatattcca ggatatcatt agagaatata gaatataaac aacaatgctt tattttcatt54841ctttcatttg caaatgctta ttacagacaa atccttggca gtgatttccc cttgtatcat54901aatattgtca taaataacat tgaaggtcta ataaatcaaa ggcggaatga agttatcagt54961ttgttgttgt tgttagactc taaaagatat tgccaaatta tagttacaac tactatatat55021agatataaat gaattaacat aaaacaaaaa ctaccttttc ctatactatg tgactattgg55081taatctctac ataaaattat tggtttttca aaatgttcaa taaagtaaaa ctagagcaac55141cataccaaca attccaagtt tttaaaataa aattattgag aaggatgaaa ttaaaagctg55201tgaactaact ttgatatagc acctggcaat ttaaaaatta ttttctgagt attatctaat55261ttggttctcc tgacaagcta aaaatgatga ttctgcctac ctcggctctc agggttgaca55321agaatactga atctcaaaga aattggtgat ctgcctgtgg tgatataaat actaagcagc55381agattcaagg gcagtgtata gctctaattt ggatcctgtt taaacaaacc aactacagta55441aaagaataca cttcatataa ttaaaatgct tttaaatttg ccacagatta ttttatgtcc55501tagaaaacag tctattttgg tgaatgctcc atgtgtactt gaagggaata tgtaatttgc55561tattgatggg tagtatattc tgtaaatacc aattagatca tcaaaccagc taggaaaata55621ttaagaggca accaaacaat tatttgaaaa atttaaaatt atgcttatac gaggtattaa55681ggctattttt aaatgagttt tatttttcag aaatatatac taaaatattt 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tactttttaa57421tgttatatta tctgWtatta aacctataga tacctttcgg taccagcttt cctaatagag57481tgagatacta caatactctt tttttctttt aattttagaa gctgaaaaaa gaagagacat57541tgctttatag atgcttattg gctgctgcac aattgtggct gaatactaaa agacataatt57601caaacaaata tttgagaaaa ctgcRtaatt caaaataatt gagttaagca tttttttctt57661ctacaattga aagactcttt tggcatcatt tagaataaag cttaattctg tttgatgtta57721cttttgtttg ttaaataata gagtggaagg gatccatatt tcaaagggca aatccatata57781gaaagatact aacttttaaa ataatccata taaaacagat atgctgcatt tatatttaaa57841ataggataac tcagatgagt catactttcc tgtaaaactt cataatgaaa taaatcctca57901ttttagtata attttgatct tattatgttt cctgtcagaa aatacatatt caattgctta57961tctccatcca gaaaagacta gagtccctga ccataataaa agcacagtga aaaagaaagc58021aaagtaccat taaagaagag gagaaagggg aagtttagaa ctctatgcca tcatgcagca58081actgtatgct ataatctggt actgaaatac agaatataaa ataacctgta attaatacca58141ctgaaatttc aaacaacttc ctgatcttta tttaaggaga aacacttccc cattctctcc58201tttctctgcc agccttagaa tccactttta agtgatgaag cacccaaacc gtttcttatt58261gaaatgaaaa cttgcaggcc acccgaagag agtgcttttc aaaatttgct ctgtatggcc58321ttagggaaca tctagagtta ctcagggtca gctttgagag agaatgggat gccccagcaa58381tcttcaatca cagcgactcc agtgctgata gattcatggt ttggttgtct acataagatt58441tcatttgaaa tgagcctttg ctaaaataaa ataataataa taataattga aatcactgcc58501ttacgaaaat atcccccaaa ttaagaagga ataatagtag ggtaaatttc aactacagaa58561ttacatctta ctccttaagg gaaagcaaaa taaaattaga aaaaacaaca acaataaaaa58621caaaaaacac agtccaaaag aaacactagt ccaaggacag aagtttaaaa atagtaggaa58681agagaaagaa ggctattaag aatcagaaat tctagcattt tctttgctgt ttgtgaaatg58741aagaaagatc aagtcagtga aaaacaggct ttctggaagt tggaaatagc agagaaacag58801ccggatctgg agtacagagg tcatcagggt agtactgtta gtttgttctt agtaacStgt58861caaccttctg cacattttct ccatttccac gctcacactc cacttctgct ttgtagtagg58921ttcctggata atcacttaat tgatttcttt gcctcaatgt cctcccttta taatatttct58981tgtaggtcac tgccatatga atttttatta cacattcttt tcaaatagtc tctctcttac59041tccaaaatag agatcatggt atattatgta gtggctaaca ttcctaggat tgtgttcaag59101tttcttttta atttagtctt aacctacttc tcgaagctga ttcttcaacc attactctcg59161gacaacaata cctttaattg tactctttct tctccaggaa cataatctcc ctttctttat59221gcttatcttc aaattaattc gagtttctgt tatcctttgt gaagtctttt gtgaaaattt59281cctctagaat gtattagaga tgcagctaag actctctttc atctgaaaca tactgttttc59341aattagttac tattttatag tgttaattct ttctatatgc tatttcttat ttagtctaaa59401ttacttcttg gtatcaccta aggtgcaggt tttaaagttt tatttttact atgacccaca59461atcagaaata tataaatatc aaaacacaga gagaatttaa aaagatgatt atctttttat59521ttttatatat ttttctttat ctatttttat tttattttcc tgatgaaaat taattttata59581agttacaaag cagttatgac ttatgctatg aaaaatactt ctctacaaaa actattcagc59641aatttttaaa gcaatttttg attaaaagaa gaaaaaatcc cagtagactc atgaaactct59701taaagaggaa acttcaaatt tccaggagaa tctgtcatgt ttaaaataca ttcgacaaat59761ataaagtaac tgaggcacaa aacatgttga cactgctaag atatcaggca gatttcaaag59821tatagaaata catagctatg attggacttc ataaaacata tatttgtatt aatatactat59881tctataaaat tgcatttttc ctttacagtt ctgtatatat tacagaataa gaaaaatcct59941aacagtgtgt ttacaatttc aacaaaccag tggcaaaaca accaaaaaag gttatataat60001ggcgactttt taaaaaactt catacctgaa tttctcctgt aattggatga gagacaacct60061ttgttccatc ggtaggctgt aatattaaag aacatatatt acattaatag atacattttt60121cacataattt atcatttcgg cttccttctg aaattagttt ctatttggtc agagcaactg60181cttggaaaat atattttatc catttggctg cttcagaata atggtgtcta cttacggttc60241tcaatacaca tttaatttta cagatgcata aatctatttt tcattttggt aagccggctt60301tgaagctgta attttatacc agcagtattt tcaattactt aagaaaatga acactttagg60361tttaaagcac ttaattaaag tatctacagg ttttaaagat ctatacagta atgtttaaaa60421tacattcaac aaataaaaga ttggtttaat acaagaactg ccgaggagca cagaaggtag60481aaaagtcctt ctttcacttc tttgctgaat gctcagcacc tccctgctcc cttccagagt60541gaatcaccca ctcaatcact ttccctataa aaattccaaa gaaattttta tagaactagt60601aaaacattcc taaaattcat ttacagccac aaaagaccct aatagccaaa gcaattttRa60661gctaaaagaa caaagccaga ggaattatgc tacttgattt caaaatctac tacaaagcta60721tagtaatcaa agcagcatgg tatggcWtaa aagcagacag acatatggaa tggaacacaa60781tagacaaccc ccaaataaat acattcattt acagtcaatt gactttcagc aaagatgtta60841agaacacata atgggcaaag gatagtctct ctaataaatg gtaccaggaa aattggacat60901ccacatgcag aggaataaac ttagatcctt atctcacacc atatacaaaa atcatctcaa60961aatagattaa agatttaaat ataagacctg aaaatttaaa attattagag agaaacataa61021gggaaaagct ctatgacatt ggtctgacca aagatttttt tggagatgac tccgaaagca61081caggcaacaa aagcaaaaat aaaaaataga agtacaacaa actaaaagtc ttttgcatag61141ccaatcagct tctgctgatt gtttcaatca acagagtgaa gagacaacct atagaatgtg61201agaaaatatt tgcaaaccgt acatctgaca gggtgttaat atccaaaata tacaagaaag61261tcaaacaact caacagcagg aaaacaaata acctgaatta aaaatgggca gagatttgga61321atagacattt ctcagaagaa gacatacaaa ggactaagag gtatataaaa aatcaacatc61381attaatcatc agagaaatac aaattaaaac cacaatgaga tatcaactca ccccagtcaa61441aatggctgtt atccaaaaga aaaaagatag tgagtgatgg tgagaatgtg gagaaaggga61501acccttgtac actgttggtg ggaatgtaaa ttgatatatc cataatggaa aacagtatgg61561aggttcctca aaaaaattaa aaatagagct accatatgat ccagcaatct cactgctgag61621catatateca aaggaaacga gatcagtatg tcagagatac caacactctc aggttcattg61681cagcattatt cacaatagcc cagatatagt accaacctaa gtgtctatca attgatgatt61741aggtaacgaa aatgtgttac atatacacaa cagaatatta tttagcctta aaaaacaagg61801aagtcctgtc atttgtaaca agatggataa acctgaagga cattatgtta agtggaataa61861ttcatgcaca gagagataaa tgccacatta tctcactcat atgtggaatc taacaaagtt61921gaacttatag aagcatagag aagaatggta gttaccaggg gctaagagaa aaggaaggat61981tggaatgatg gtcaaagggt acaaacttct cttagaagga agtcaaaagc tctgttatac62041aacatggtga ctacagttaa tagtaaatgt atatattctt caaaattgtg aagagagtag62101atttaaaaaa aattttttta agccaaatat ttcagaggaa tgaagagtag atttcaagtg62161ttcggaccac aaaaaaataa gtgaggtaat ccatgttaat tagctcaatg tagccattcc62221acaacgtatt catttttttt ttttttttgg agacagagtc tcactctgtc ccctccggct62281agagtgcagt ggtgtgatct ctgMtcactg caactccgcc tcccatattc aagtgattct62341cccacctcag cctcctgagc agctgggact acaggcatgt gccaccagac ctggctaatt62401tttgtatttt ttggtagaga tggggtttca ctgtgttggt tagggtggtc tcgaactcct62461ggtctcaagt gctctgcctc tcctggcctc ccaaaatgcg gggattacag gcatgagcca62521ccatgccagg ccacaatgta tacatatttt aaaacatcat gctgtacagt ataaacatat62581acaagttttg ttaattaaaa ataatttaaa atttaaaaat ctaagcaata ttgcatgaga62641tggttaattg cttaatttaa aaagaagttg tcttcaaata tgttcaaatt gtatttttat62701aacatagaaa catctacaaa acagaattta gtattaatta tgttggtatt ttagttggct62761agtttatgaa ttatttttaa atgattatca aaatagagcc tctataaagt aattattcaa62821aattagcaca taattgcttg tggtaaaatg tagttgtgtt aaaatttatt ttaaaattta62881gcctaactgc ctttgaaatt actttgctta tgaagctttt ctatatttag tttcaaacaa62941atcattttac gatgcaatga ttaaggtcaa cctacttttg ttgtttgctt ctgaataaaa63001ggattctttt tttggtgaca tctgctgtat ttttatagtt aaaaaacaaa ctttattttt63061tatgtctggt cacttgacct aagcatatac agcctgtcct tcccatatat ctcaccacat63121aaagacaaac ctttattcat ttttaaacaa aaggacactt ttttcgcatg tatttatcaa63181ctacagggat agtttttctt aagaatcaaa tctatgaggg aaatcataaa ttttgtatat63241ttcctctgct ttttataagg ttaaaaaata attttatgct tttttaaatt 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ttgtttataa64981ttatataaat acataaaaaa atctatccat aactttatat ttgtctatat ctgtgtttNt65041gtctacatct tcatctacaa ttacatctgc ctctgcaatt acttctattt ctatctcttt65101atttacagtt gaaactagat ttcttttacc taccagcttt tggtggctca tgaagttcca65161gatgctggga attttcagaa tctgatagca tattgaggtc cctaaaaatt aaaacaaaac65221attaagaaag atattgagga aatctgagac aaagagcact tttttccaac tacccttaat65281ttttttcata ggtaaatgag aagtggttaa tgatataaag caactccaaa gcatgcctac65341catcctaacc caatatatat ttgcaatgca gttaaaagtc agagctgata aaaatgatta65401tttaaggtga aatctttatg aattaaatgt aagttcattt ttaaatacat atccatgaaa65461tctaaatggg ttcaggcttt catttaaatg aagcatgttt ccataaaagc atgaaaggga65521agaggctgtt gacacaatca gaaactaagt tagctgaatt ctttctatgc tttaggcttt65581gattcacttt attttataat tccagaaaat cttgctaact ggcaaaggtt ccaccttaac65641aattttaaat taaaagagtc atggatagcc aaaaaatgga aacagaaaga tggggaaaaa65701ctcacagtct tacacatatt tctgcttccc cactttgctg actaatgata ctctgaactt65761cttcatatgt tttagaagtc aagggaattc cattccattc caatacttgc 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gaattctatt78421agataaattt aatagagtat gaaataacta aaaggaatca agcagaaatt ctggaggtga78481aaaatgcaat agatgcaact gaataatgca tcagaggtct ttaatagaag aattgatcaa78541gcaaaagaaa ggattagtga gcttgaatac aggctatttt aacgtacacc aagaagacaa78601aaaaagagag aattttaaaa ataaagcaca cctacaggat ctaggaaacg cctcaaaagg78661gcaaatctaa tagttattgg ccttaaagag gaagtagagg aagagattgg ggtagaaagt78721ttattcaaag ggataatgac agagaatgtc ccaaacctag agaaatatat caatatccaa78781gaacaagaag gttttagagg gccaagcaga tttaacccaa agacaactac ctaaaggcat78841ttaataatca aactcccaaa ggtcaaggat aaagcagcaa gagaaaaaaa aataacatac78901aatacagctc caatacatct ggtagcagac ttttcagtgg aaaccttacg ggccagggag78961agtgatatga cttatttaaa atgctgaaga aaaaaatctt ttacgttaga atagtatgtc79021tggtaaaaat acccttcaaa catgaaggag aaataaatac ttaacccaga caaaaaaaag79081ctgagggatt tcaataccag agctgtccta caagagatgc taaagggggt acacttcaat79141cacaaagaaa aggacagtaa tgagcaataa ataatcacct gatggtccaa aattcattgg79201taatagtatg tacacagaga aacacatatt attataacgc tgtaactgta gtttgtaaac79261tactcttatc ctaggtagaa gattaaatga tgaaccaatc aaaaataata actacaacaa79321cttttcaaga catagcataa taagatataa atagaaacaa caaaaagtta aaaagtgggg79381ggatgaaatt aaggcataga gtttttatta gttttctttt tgcttgtttg tttatgcaaa79441tagtgttaag ttattatcag gttaaaataa tgggttataa gatagtattt gcaagcctca79501aggtaacctc aaaccaaaaa gcatatgata tatacgcaaa aaataaaaag caaaaactaa79561attatatcac tagagaaaat catcttcact agaggaaaac aggaaggaaa gaaaggagga79621agagaagacc agaaaataaa caacaaaatg gcaggagtag gctctaactt atcagtaata79681acattgaatg taaatggact aaacacttcg atcaaaagat atagagtggc tgaatgggtg79741aataaacaag acccattgtt ctgttgccta caagaaatac acttaacctg taaagacaca79801catagactga aaataaaggg agggatgaag atatttcatg ccaatgtaaa ccaaaaatgg79861gaacctagaa attaataaca ggaggaattt tggaaactat tcaagtacat ggagttatta79921cacttagaaa aagtagattc caagacaaaa gctataagaa gagacaaaga agttcactat79981ataatgataa agaggtcaat tcagcaagag aatataacaa ttgtaaatat atatgcaccc80041aacactggat tacccagata tgtaagggaa atattgttag agctaaagag agaggtaggc80101tccattacaa taatagctgg agacttcaaa accccatttt cagcagaaaa tcagacaaag80161aaacatgaga cttaatctac actatagacc aaatggatcc aatagatatt tatagaacat80221ttcagccaag agctgcggaa tacacattct tttcctcagc acatggatca ttctcaagga80281tagaccacat gttaggtcac aaaacaagtt ttaaaacatt caaataatgg aaatcaccaa80341gtattttctc ttactacaat gaaataaatc tagaaataac cagaggaatt ttggaaagta80401ttcaaatata cagaaattaa acaatatgct cctgaatgac cagtggatca ataaagaaat80461ttagaaattg taaaatttct tgaaacaaac gataatggaa acacatacta aaacccatgc80521aaaagcagta ctgagaggga agtttatagc tataagtgtc taaatcaaaa agagaaaaaa80581gttgaaatga aaaatctaat gacgcatctt aaagaactag aaaagcaaga gcaaaccaaa80641ccccaaatta gtagaagaaa acaaataata aagaacagag tggaaataaa tgaaatcaaa80701atgataaaaa tacaaaagct caacaaaacc aaaaattggt tttttgaaac agatatatga80761aaaggtgctt aacatcattg atcatcagag aaatgaaaat caaaactaca acaaaatata80821atttcacccc agttaaaatg gcttatgtcc aaaagacagg caataacaaa tgctgatgag80881gatgtggaga aaagagaacc cttgttcact gttggtggga atgtaaattg ctacagtcaa80941tatgcagaag agtttggagg ttcctcacaa aactaaacat tgatttatca tatgaaccca81001caatcctact gctgggtata tatccaaaag aagggaaatc agtttattga ggagatatct81061gcactcctat atttattgct acactgttta caacagctaa gaattggaag caaccagagc81121aagactctgc ctcaaaaaaa aaaaaaaaaa aaaagaattg gaagcaacct aactgtccat81181caacagatga atgaataaag aaaaggtgac atatacacaa tggggtacta ttcagccata81241aaaaagaaca agatccagtc atttgtaaca acatggatgg aactggagat catcatgtta81301tgtgaaataa gccaggcaca gaagacaaac atcgcatgtt ctcgattatt tgtgggatct81361aaaaatcaaa actattgaac tcatggatat agagagtaga aggatggtta ctagaggttg81421gaaatagtag tagggagttg gagggaaggt ggggatggtt aaagggtaca aaaaatagaa81481taaataagac ctgctattcg atggcacaat agtgtgacta tagtcaataa taacttaatt81541gtacatttca aaataaccta atgagtataa atgtattgtt tgtaactcaa aggataaatg81601cttgagggca tggataccct attctccatg atgtgtttgt ttatctttgc atgcctgtat81661gaaaacatct catgtacctc ataaatatat atacctacta tttatccaaa aaattaaaaa81721ttaaaaagtt atactttgct tactttgctt atattggagt caattatact acatgattat81781tatgaatgat ataatcaaaa ttaaaagggt aaagttgatg cacaatttct ttcataatta81841gaagtgtcaa aagatgtaaa taacatagag aagtcagttc aaacaacaaa aatatcagta81901acaattttat catcctgaga gaatcttcaa agaaataaaa ttatgtgtca agttatttca81961taaaaagtgc aaatattctg ttctttatat cacaatgaag aattagcaaa taattcactt82021tgacttggta gatgtttaat aaataacata aaaaatcaaa tgtcatttga tccctgaatt82081ccatttgaaa agttatcaat ctctttaata aatagaattc aatttcaaat gttatcaatg82141ttagagtgta tacaatatcc tgcaataatt gattttccat tgaaaaataa gaattgacta82201agaagaaaac agtttaacag caatttctgc caactgaata gaataaaact taactatgtt82261ccctccagct ttctaatatt catgtagaga aattctttct ctaattgtaa ataggattac82321aactgtaaaa gtaatggcaa tatagttcta tatgaattat agagggtttt catatgtata82381ccatctgatt ggtggttatt tcagttaaaa ggagtaaaat ttaacccatt tcacttgtgg82441taataccttc ggtattaata aggagcacca atacaggact cacacaaaaa agttattttg82501cccatttttt ttctcatttt atcttcacag caatccagta aaatagatag ggtaagcaaa82561tccccatttt cacaataaaa cccccatagt tttagaaacc acatttggtg gcagtttgaa82621aagccaggtc tcctcattcc tcatctaagt tattttggta ttataaaggt ttttttaatt82681tttaagttat acagaatatc taattataaa cttctggcta ttttatagct aatatcttag82741atgtaagtgc ataatcagta taaataaata tattgaatta caataaaaca tcacatcaag82801tccacaaata tttagtcagt aagtgccaaa gagaatacta aattctgggc cacttatatc82861aattagttat aatttctgac atgattactc ctactatgga gtaggaacat catgaacaaa82921tgctcacaga aggcaaataa catcatggat ctgggatcag agtggggctg gtgggtgtat82981agatgggagg tgagaatgga taaagatgag tttgtatagc tggaatgtgt tgtggaaggc83041ttacagctaa aattttattt ggacttaatt aattttctaa tattaagaca taaaaggaaa83101ggtttataat aattagatca ttattttaaa aacatcttgc catttggaga ggatgtatgc83161atttctcctc caacttcagc aggttagttt cagtttcttt tgtgttttct ctgtttcttg83221tacacacttc tgttatcctc aaaacgttat actttaattt attcagttat gtaattgcct83281ctattttgac aatggagaca ctagtgaatg acaatttgct gaatgaatga atggataaaa83341tcagagagac tggaagcccg gagaatggtt aagatttaaa aaaaataaga caggaattca83401aaattgcaat tgattattaa ttcttacatg aaataaaatt ttcagaaaat attgtcatga83461tgaaaaaata ggactgctga gaatcagttc ttggaatcaa tatagtactt aaattcattt83521tttccacagt ttattaatgc aaactgtcta agcactctaa aRtattgaga gattttattc83581acctttgttc tcataaataa agtgatattg atacagccat aattttctta aataaagaag83641tattacagaa tgattttcct tcataaacat taaaactgtg gctcttggct gccttttgtc83701ctactgagat gcaaattaca aatgatagaa ataaaaaaaa aaattgaact gccacctgta83761gaaccacaca aatgccgggc atggcggctt acacctggag tcccagcact ttgggaggcc83821aaggtgggca gatcacctga gatcaggagt tcgagaccag cctgaccaac atggtgaaac83881cccgcatcta ctaaaaatac aaaaagtagc caggcatggt ggccagcgac tgtaatccca83941gctactcagg aggctgaggc aggagaatca cYtgaaccag ggaggcagac agtgcagtga84001gctgagatcg cgccattgca ctccagcctg ggcgacagag caagatgccg tctcagaaaa84061aacaaacaaa caaacaaaca aaaaacMcac aaatatacat ttatatggta tatttcctac84121taaaatatat ggatttattg tgttaattta aaataagatg caagacttag atacaaaata84181ctctgttggc ttgctgtatt atgcagggat ttaagttttt aaaaaatggt gaagggattt84241tggcttataa atatgaatac ctaataaatg aatgaacata ttgagatcat tttcattaac84301taaattaaat acacatattg tgttgataca tatgtatatc tattatacat atatataact84361atttcttaac acctccctgt tcccaccttg gtcttaactt acataggttc ttaccaactc84421tccttgcttc tactctttac ctctttaaaa ctattcttaa tacagcagta agagtgatcc84481tatttaaacg tcattcagct tatgtcaggt caattgcgtc tcatctcaca taagtacacg84541ttaaagtcct tacaatggcc tgcaagattt gacccctgct acttttggta tccgctcttt84601cacgtgttca ctcagtttca gccacgttgg cttcctttct gtcttgctgt tcttggcaca84661caacaagcat gctcttgctc aggatatttg tgtccttgtc acttctctgc ctataatgca84721agacaactcc cagatcattc tttcccctcc ttggtggctt tactgaattt ttgcctactt84781gccaaagctt tctcttacca cctcacctca aattgcaacc acatcctcct acgtgccata84841cccaccctcc ttgctatatt tttctcctta gcgtttatta tcgtctaata tatgatactt84901aatatattat atgagatata tttaacattt atatatgata ttaatatttg ttataatctg84961tcttagctat tagaatgtaa gacccatgct ttcttcactg aagccctcac atgtccaaga85021agagaaatta ctttatacaa tatgtactaa aaaaaacttt gaaaaaatga atgaataaat85081aatggcattc agcaaactca agtgctgtgg ctgacagtga ctacgatttg gggaatgagt85141agagtgtaaa aggaaacaaa agtagcttaa gagaaaaata aaggaatctt attttccttc85201actactctca atgaaatttt atacacacac acacacacac acacacacac acacacacac85261acaccccttt aatatgaatt tcttgaggac tgaagtaaaa acctgcattc ttgtaggcta85321ttgtttccac ttcggcactt atcattgtat ctggaatagt actccataac tatgctgaac85381tattaaaaat taattaaaat aaatgagaac cttaaaagat ctgtatagtt cctggacagt85441gtcttgccct caggagctta tagtctgctc ttagaaacta cctgaaggtt agttggaatt85501ctttcgagtg ggggaaaaat ctgtaagctc catttagtca tggtgttggt tctaatgcaa85561gcagagtcaa ccaatgtatt ctgcatttct ttatttctgt attttctttc aatgaaacta85621aaagaataca tatatcaaaa ttaaaaggaa gacaaacatc attaatttta aaaaaagagt85681tttggtccag gtgcagcggc tcaggccttt aatcccagaa ctttgggagg ctgatgcagg85741aggatcgttt gagcacagaa ggtcaaggct agcctgggca acatggcaac actcctgtct85801ctacaaaaaa tttaaaagaa ttagcatatg tggtagtgca tgccttttta ctacttacaa85861tattctaagc atgacattat gtttcctata tcttattcct aaatttgata acaattttgc85921caactattat cccWgttatc cagctaagaa cacctgggta ctaaaagttt taaaaaatcc85981aggcaataag taccagatct gaatttaaat cctgatctaa ctctaaaagc atggctgtct86041acaaaccact ggagatacta aagcattggc tggtggaaga tctggtatgg atattaaaaa86101gagaggttag catccttgct tccaaagtaa ccactccaag cttacgaaaa agtgcagtct86161attccaagaa atattggtta gacaaatccg gaaatctaac tgtaggaaca attgttcctt86221atgaagcaat atcactgaac acagaaatac ctcaaagtat gtcaaccttg atgaaggcaa86281tcaaattagg atttggggca ttgtaccaca taaaacagga cattatttag tttccttcat86341gtccctatca caaagattat aatgtaagca taattctcct ctgataaatt tcaaaataaa86401tggataaaga agagaaatat gtttctttgt actggcaatg tatctaatca ctagtatcgt86461gatgtggtgt gatatcacta catcttgaaa ataatatatt agaacctatc agcccaggga86521ccagtaattt gtatttttct tcacagaaga gagtttccta aatatatatt agaaaatctt86581catatcaaca tgaWatgtgg tctgtgtcat gtgtacctaa cgcaccagct ttaaggaatt86641ttgagaaatc tcatgctaat gaccaatccg tacctggtaa tccccctgga acaagaacaa86701ggaagctcac tcatctgtgt gctgtgtttc ttgatctcca accttgatca ctgccatgcc86761atctgtaaca tatgccaaaa aagcataagc agaaatagag actttcttgc ctaggagcat86821tcttccttat gttatgctgg agtgcgtcct ctgtcctcta gtaatgctac tggaattagc86881tcttctacca aagacttttt catgacaagg aattccagag gccaacttct aaaatcaaaa86941ccaccactcc aaaaacagct aatttcatcc ttacttcaat ccctgattct cagcattaca87001tccaacagca ttttcttgcc agaaaccctg tcagttcatc tttaagaaga agagcaagct87061agctggacca atttcactca cctactctga aacaactaaa aactaagaat ataaaagtat87121gcatgaacta taattcatgt cacagaggca tttcctcaca gaatacctat ggacaaacaa87181tgaagtttct ctcatttcat tacaagtctt atggccttgg acgtgcttac ttgccatgta87241gctctggcta aatctataag caaacatagg atatcactgg ttttagcttt gtgctcttcc87301atgctgtgtt accactgatt cattaagaat gttcttgaag acacattcag attttaatgt87361tatatttcta tcttagctcc aaagagtttt acaaaaattt tctaaattta tcaattataa87421taatatagat acaagttatc ttcaaaagat tttattaaat taacaaagtt gaacaactta87481tcaatataaa caattgtttt aggttcattg gatatatgat atttagtcct tgtggattca87541ttgttctctt acaaaaattg ctgaagtttt aaaagtatca cgatggtaat gtatttcaca87601attatgttat aacacagaca actaggatga aaattaattt ggagaactag ttcaaaaaat87661ccagtgatta aataaagctt agaaccaatg atatgtatat aaaagtttag atttagtcca87721tgatattaac caagtaaaat tgattttatc tatattttct actctacctt ttctggagat87781ataatattgg cataaattga taaaaccata taatattgtt aataagttta acaattgcta87841tggataattg ttttaatgtc ctaactcttg ttattgtatt aattcagaat acaatgacta87901agtctctaag tagagcaaaa acctgcattg tgaacttatt gaagactcaa ctggacatga87961ttatatgtct aatttttttc caaattaatc agtctcttca atgaaattac tatcaaatct88021caacagagat tgtcatgaaa attgaaaaac agattccaga atttatatgc aaaaccaaag88081ggtcaagaat tgttccgaga gcatcctgac aaacggctct aggatggatc cacctacgaa88141atgccaggat ttaatataaa gatgttacaa ttaaaacagt gtgatatcgg tgtagggata88201gaaaaataga ccagaacaga gaacctggaa aagatctgtg catgcctttt aaaacttaat88261atatgacaga tgggactagc agatcagtgt gagtaataat acactattca aaaaatggtg88321cacagtagta gtaaagaaaa ataagaaagc aatcctattt aaaacagcta cagtataatg88381aaatacctaa gaataaattt aactaaataa atgaaagatc tttacaatgg aaactataaa88441acatgaaata tcttttcatt cttatgtgtc ctcttcaatt ttttcaatgg ataaaaaata88501gaaaggacac ataaaaaatg gaaagatatt tcatgttcat ggattgaaaa aattaatatt88561gttaaatgtc cacactatgc aaaatgatct gcagattcaa tgcaatctct atcaaaatat88621caatcaatgt cattcttcac agaaatacaa aaaaatttct aatagttgag tggaaccaca88681aaaggccctg aatagacaaa gcaatcttaa gtgaaaagaa caaagctgga ggcatcacac88741tgcctggctt caaaatatac tacaatggta tactaaccaa aagagcatgg catggcataa88801aaatagacac atagactaat ggaacagaat acagaaccca gaaataagta catgcattta88861cagccaacac agtttcaaca aaggcacaaa gaatatacgc tggcaaaagg atagtctttt88921taagaaatgg tgctgacaaa actggatatt catatgcaga agattaaaat tagagcatta88981tctcttacat atcaaaataa actctaaatg aactaaagac ttaaatttaa gacctgagct89041tatgaaacta ctagaagaaa aacattaaaa gaatactcca ggacattggt atgggcaaaa89101atgttttgag taagacctca aaagcacaag caacaaaagc aaaatttgac aaatgggatt89161acgtcaacct aaaaagcttc tgcatggcaa aggaaacaat caacaaagtg aagagacaac89221ctacagaatg ggagaaaata tttgcaaact atccatctga caagggatga ataggtagaa89281tgtataagga actcaaacaa cttaatagca aaaaaaaccc caaataattt gatttaaaaa89341atgagcaaaa gatctgaata tacatttctc aaaagaagac atacaaatgg ccaacagata89401cattaaaatg ctcaatatca ctagtcatca gggaaatcca aattaaaacc acaRtgagat89461ttcacctcac cccattaaaa atggctcttt tcaaaataga gatgctggca aggagatgag89521aatagcagat gctggcaaag acttgaagaa agagaaatgc tcatacactg tttgtggcca89581tgtaaattag tccaaccact ataaaaaaca gtatggagat tcctcaaaaa accaaaaata89641gaactaccat acaatacact gatcccactg ctaggtatat atcaaaagaa aggaaataag


Following is a REPS2 genomic nucleotide sequence (SEQ ID NO: 7).

>X:16315151-165284501ctctgtcacc caggctggag tgcagtggtg ccatctcagc tcactgtagc cttgacctcc61caggctcaag tgatcttccc acctcagcct cccaagtagt tgggactaca ggagcacacc121accatgcctc gctaattttt gtagttttta taaagatgag aactcgctat gttgcccagg181ctggtctcga actcctgggc tcaagcaatc catcggcctt ggYctcccaa agtgctggga241ttacaggtca ccatgccagg ctgagacagt aacttgtaag atttaattgt tggtcgggtg301cagtggctca cccatgtaat cccagcactt tgggaggctg aggcaggtgg atcatgaggt361caggagttca agaccagtct gaccaacatg gtgaaactct gtctctatca aaaatacaaa421aattagctgg gcgctgtggc gggtgcctgt aatctcagct actcgggagg ctgaggcagg481agaattgctt gaacctgggt ggcagaggtt gcagtgagcc gagatcatgc cacggcactc541cagcctgggt gacagagtga gactctgtct caaaaaacaa acaaaaaaag atttaattgt601cattagtggt ttgtggggca cataagggtt caagaacttt aacccctgtc tttcttgatg661agtttagaac tacctgggta aggtccatgt gcacatcaac cttgttgttg ctagacttgg721gttggagatg atgattagga ttgcagccac actcaggcta cttcttacat attctatccc781cttacttccY ccacctcaga gtctataaaa cagacctata cctgggaagc tgcWaggaaa841ttgctgtctt ctaatcatca agtcctgcag gtggtaacag agaggaccca aattacgtgt901gctttctaga tggaatcggg gttagagtgg gggtagagtg gagtcccctc tgagggcaac961agaattcaga ggtggtagta ggagagggag gtaggagaga gagaaagaaa gagacagaga1021gagagagaga aagaatatta gcctgttctc acactgctaa taaagacata cctgagactg1081gataattcat aaaggaaaga ggtttaattg actcacagtt tcacatggct ggggaggcct1141cacaatcatg gcagaagttg aaggagRagc ataggcacgt cttacgtggc agcaggcaag1201agagcttgtg taggggaact gctctttata aaaccatcag atctcatgag acttaatcac1261tatcatgaga agagcatggg aaagactcac ccccataatt caattacccc ccaccaggtc1321cctcccatga cacgtgggaa ttatgggagc tagaattcaa gatgagattt gRgtggggac1381acagacaaac catatcatgg aggaaaagag agggggagag agacagagag agagagaatg1441aaccaagaat gatcaatttc ttctcccttt catcacttct ctttccttag agaagagtga1501gggggactag gagaagtgcc aatgttactt catcaatatt ccttcctgag gagtagagag1561gaagctttct catgaaaata atgaggccag gggaatagga ttctctctca cactgttaaa1621gatctgcctt ttttcactaa tagcagaaaa aaatattaaa atcattcaaa caggtgatta1681tttagtgttt atattggtcc atcccttaat aaataccttt caaaattcac caatctttga1741gatacactca atagtacagt catgcgttac ccaatgacgg gtatgttctg agaaacacgt1801tgttgggtgt cttcgttgtt gtgtgaacat cacagggtgt acttacacaa atctagatgg1861tacagcctac tacacaccta ggctatgtga aaattgtcag aatcaaaatg gagtcgcttg1921tgttaaaaaa aaacacacaa aaacctgaca aatagagctg aggaaggcta cgaagagagg1981gttctcacac ttgtatgcct gataacaaaa ttgcaacaaa agactgcaaa aactgcaaac2041ttacacaaat gtcatcacaa ccttctgtaa agacatctga ccagcaactg cctgtctaaa2101cttgaactgg tgccatcctt gttattgatc cttgtcacca aaaataacca tttcaaaaca2161actatgtaac cttcattttt ccttgaaaaa tttttgtctt cctttacctc cacgaatatg2221cacaagttta ctgtggcata catattccca ttgcaatgcc ctattcccac atgaacaaca2281ttttcttttc ttttttcttc ttcttttttt tttttcttta gacagagtct ccctctgttg2341ctcaggctga gtgcagtggt acaatcacga cttactgcag cctcagcatc ctgggctcaa2401gtgatcctcc cacctcaggt tcccaagtag ctaggactac aggtgcaYga cacaatgccc2461cacacccagc taatttttgt atttttttgt agagacaggg ctttgccatg ctgcccaggc2521tggtcgtcaa ctcctgggct caaatgatcc acccacgttg gcctcccaaa gcactagcat2581tgcaggcatg agccaccgtg cctggccata aacatcattt tcttttagag agcctctctc2641tgacatttaa gttgacagct atatggtaca gcctattgat cctagactac aaacctgcaa2701gcacagcatg attgtattag tcaattctca cactgctgta aagaaatgcc tgagactggg2761taatttataa agaaaagagg tttaataggc tcatggtttt gcaggctgta cgggcagcat2821ctcttctggg aaggcctcat ggagctttta ctcattgtgg aagacaaagt gggagcaggc2881atcttacatg gcagaagcag gactgagagg ggttgtgggg aggtgccaca cacttttaaa2941ccaccagatc tcatgagaac tcactcacca tcgtgagaac agcaacgagt ggaaggggct3001aaaccattca tgaaggatcc acccccatga tccaatcatc tcccaccagg ccccacttcc3061aacactgagg attataattg agcatgagat ttggatgggg acacagagec aaaccatatc3121attttgcccc tagcacatcc taaatctgat gtccttctca ccttgcaaaa tcccatcatg3181ccttctgaat agtccctgaa aatcttaatt cattccagca ttaactcaaa agtccaatgt3241tcaaagtctc atttgaggca aggctagtcc cttctaccta tgagcctgta aaataaaaaa3301caagttagtt acttctgaga cacaatggag agataggtat tgggaaaata tacccattca3361aaaaaaaaga aattggacaa aagaaagggg ctacaggcct catgcaagtt tgaaacccag3421cagggcagtc attaaatctt aaagctccaa actaatctcc tttgacacca cgtcttgtat3481ccaggacaca ctggtgcaag aggtgggctc ctaaggcttt gggtagctcc acccctgtgg3541ctttgcaggg ttcagccccc atgacagctc tcatgggctg gcattgagtg cttgtgcctc3601ttccaggtgc agggtgcaag ctgttggtgg atctaccatt ctggggtctg gaggatggtg3661accctcttct tacagctcca ctaagcagtg ccccagtggg gattctgtgt gggggcttca3721accccacatt tcccactctg cactgcccta gtagaggttc tccatgaggg ctctgcctct3781gcagcagtct tctgcctgga atccaggctt ttccatacat cctctgaaat ctaggtggag3841gctcccaagc ctcaaccctt gcactctgca cacctgtagg cttaacacga ggtggaagca3901gccaaggctt gtggcttcta ccctctgaag cagtggtctg agctgtacct gggccccttt3961gagccagggc tggagcaaga gcagcttgga tgcaggaagc aatggcccaa gggtgcacag4021ggtggcaggg ccctaggcct ggctcaggaa acctttcttc cttcctaggc ctctgggtct4081gtgatgggag aggctgcaat gagggtctct gaaattcctt tgaggccttc tccccattgt4141cttaggtgtt agcacttggc tcctttttac gtatgcaaat ttctgcagcc tgctttaatt4201cctcccctga aaatgggctt ttcttttcta ccacatggcc aggctgcaaa tttttcaaac4261ttttacactc cgcttccctt ttaaatgtaa gttccagttt caggtcattt ctttgctcac4321atatatgaac ataggctgtt agaagcagcc aggccatatc ttgaacctta gaaatgtctt4381ctgccagata ccctaaatca tctctctcaa gttcaaagtt ccacagatcc ctagggcagg4441ggcacaatac aacaaagctc tttgccaaaa tgtaacaaga gtagcctttg ctccagttcc4501cagtaagttc cttatttcca tctgagacct catcagcctg gccttcattg tccatatcac4561tattagcatt ttagtcataa ccattcagtc agtctctagg aagttccaaa ctttcccaca4621tcttcctgtc ttcttctgag ccctccacac ttttccaatc tctgcacgtt acccagttcc4681aaggtcactt tcacattttc aggtatcttt atagcaatgc tccactccta agtaccaatt4741ttctggaaaa aaaaaaaaaa accaaacaaa aaaaacataa tgtaaagaat atctctgcaa4801tgggaataat tggattccat gagaaacaat aaaaaagaat atctcttgat gtctcttggc4861tatttctgaa aatatgatcc ttctggcttt tctggccatc atggagccaa aacaagtgtg4921taagctttga accacaaaaa agttcttgtt tactgtcagc cttaagggat atgagtgctt4981ctagtttcta aattcttatc tgtgtttatt ttgaagtttg ataactattt cttctattga5041aaaaactttt agaaataaag aacagtactt ggcattcctt agatattaac atatttttac5101aagtgccatt tttttttcct cctgtgccct gggtattact gctgttgggt cccctttggc5161tgtgccccac atcttttgtt gagcagcttc tttcttacat cctggagacc ttcagagctt5221ccctttcttc tgacccagcc tgctgtccct cactcattat ttccaggttg ccaagtctct5281ggggctcctc Yctttctgtc gtgtcctttt acctccccag ttccacccac tctcagtcaa5341cagccatctc agccccactg atcaagcctt cctggctgag ctctggctca tgaattttct5401ctcttgtttc aactcccatg gcagttaacc cagagtatac agtttagcac ttaattgtat5461attttctttc atttattttt atttgttgtg ttagttttgt ctcttccagt gaggcttcaa5521actccttgaa agcaacacat cttctattaa agaaaaattc ctcttagtac tctggtaggg5581atacagtaaa tactctcttt tctatcaata aatgtcaatt gtctcactga tagattgata5641ggcacagatg tgagactttc tacctgtctc cgtcatatct ggtgcttaat aaatatctgt5701caaatttatt atacttagtt gagtcataaa gatcagttct gaatctggta ttcagtggga5761caaatcacac tgtctctgca tccattttct catctgcaaa atggagatta aaaatgtctt5821cccaatctac ctcagagagt tactatgagg cttaaatgag ataatagtta caaaagtgct5881ttgaaaagct ttgcagcttt tttcctctac tgcatgaaga ttagggatta ttatagctca5941catttccttt gacagtagtc acagggtaaa gtagggtttg gtgtttatca cacagagtgt6001gtccctttct agcctttatt agaatggttg gccattctca gggaaacctg agtacaacca6061gaaaggtagg aagagaataa acatctggct tccagttttc ctgagaaggt atcacagcta6121tggaaactac aaaggtggtg aaatttcttt tcttttcttc ttcttcttct ttttttttaa6181gacagtttcg ctctgttatc caggcgggag tgcagtggca cgaactcagc tcactgcaac6241ttccgcctcc caggtccaag agattcttgt gcctcagcct cccgagtagc tgggattaca6301ggtatgagcc atcacgccca gctaattttt gtatttttag tagagacagg gttttgccat6361gttggccagg ctggtcttaa actcttggcc tcaagtgatc ttcccatctt ggcctcccaa6421agttctggga ttacaggcat gagccaccat gcgtggcctt ctttcctttt ttaaattctt6481tttttctttt taaattttaa atagaagcag ggtctcatta tgttgctcag gctggtccgg6541aactcctggg ctcaagtgat cctcctgcct cagcctccca aagtgatagg attgcatgca6601tgagctacca cgcctggcca ggtggtgaga ttttgaaatg gaaattccca agtgaagtta6661gtctttattg ctcctaattt acatatcctt tctgcactct tgttccattc cgaagctttc6721tgttgtttta gttccctcct ggcaggttgg atgcaagacg tggtgtaaag aaagcttagt6781gaagaggtga gaacttcttc cttctggaga agatcctctt ctctgttcca tttttccaga6841ttccaaaagg aaaatattac tctggttatc actgaggttg acatcatttt ttatttttct6901tcttcttatg tatctctctt cactctctcc ccctttcctt tccttctttt tgtatttaca6961atttcatgat tttttcccct tctcctctct tcctgacctt ggtcctttag ttgaccataa7021taccaaagta tatttgttat cgtcagtggt atttcttgag atgttttttg tctctgtttt7081taataaaaat aaaactattt ggggtgtaag ttttatatat ccctttatta aagcaaatta7141taaaatgtat taattttttt attgatacat aataattgta catatttacg gggtatatgt7201gatattttgg tacatgtata caatgtgtaa tgatcaaacc agaataactg gcatatccat7261aatctcaaac atttattatt tctttgtgct cggaacattt caaatcttct aggtattttg7321aaatgtacaa taggctgggc atggtggctc atgcctgtaa tcctagcact ttaggacgct7381gaggcaggtg gatcacttga gccctaggag tgggctagcc tgggcaacat agcgagacct7441tgtctctata aaaaatacac aaattagctg ggcatggtgg tgcacgccta tagtcctagc7501tactcaggag gctgaggtag gagaattgct ggagcccagg aggtcgaggc tgcagtgagc7561cgtgattgca ccactgcact ccagcctggg tgacagattg agaccatgtc tcaagaaaaa7621agaaatacac aataaactgt taattatact caccttactt tgctatcaaa cactagattt7681tactccttct aactgtattt ttgtatccat tagctaacct cccttcgtcc cccagcccac7741ctttcccagg ttctgataac tattgctcta cctccatgag atcaactttt tcagctctcc7801catatgagtg agaacatgtg atatttgtct ttctgtgtct ggcttatttc acttaacaca7861atgaccttca ggtccatcca cgttgctgca aatgacagga tttcattctt ttttatggct7921gaatagtatt ccattgtgta tatataacac attttcttta tccatgcact attgagggac7981acttaggttg atccatatct tggctattgt gaatagtgct gcaataaaca tggaggtgca8041ggtatctctt tgattactga tttactttcc tttgaataaa tgcccagtag tgggattgct8101ggatcacata gtagttttgt ttttagtttt ttgagaaacg tccatactgt tttctataag8161ggcagtacta atcgtattct caacagtgta aaagactttc cttttctctt atatatggag8221tgagatgata tctcactgta gttttgattt gcattttcct gatgataaat gatgttgagc8281atgttttcat atacctgttg gccatctgta tgtattcttt tgagaaatgt ttatgcagat8341cctttgccca ctttttaatg ggattattat tatttttttg ctgttgagtt gagttccttc8401tatatgctgg atatcagtct cttgttggat gaatagtttg caaatatttt ctcccattct8461tcaggttatc tcttcatact gttaatgttt cctttggtgt gcagaagcct tttagtttaa8521tatatttcca tttgcttatt tttgtttttt tggcctgtgc tttcaaggtc ttagccataa8581aatccttgtc cagaccaatg tcctgaagca tttccccaat gttttcttcc agtagtttta8641cagttttagg tcttatgatt aagtctttaa tacatcttca gttgattttt gtataggtga8701gagagacata aaggtctagt tttattcttc tgcatatgga tatctggttt tcccagcacc8761atttattgga aagactgtcc tttcccaatg tatttccttg gtggtttgtt ggaagtcagt8821tggctgtaaa tgtgtgggtt tatttctggg ttctccagtc tgttctattg gtttatgtgt8881ctcttttcat gtcagtacca tgctgttttg gttactatag ctttgtagta tattttgaag8941tcaggtagtg tgatgcctcc agctttgttc tttttgctca ggattgcttt ggctattcag9001ggtcttttgt ggttttatac aatttttagg attgtttttt ctatttttgt gaacaatgtc9061attggtattt tgatagggat cacagtgaat ctgtagattt ctttgggtag tatggtcatt9121ttaataatat taattcttcc aatccatgag catgggatgc ctttccatgt ttttgtgtcc9181tcttcaattt tttaaatcaa tgttttgtag ttttctttgt agtaaaatat attttttatt9241agtgcttaac cctagactga tgttttcttt tgggttattg tttcccagtg aggcttcatg9301catatatttc tgcaggtctt cccaggaaaa atgacattta aaaagagaat ttttggaaaa9361gagatgcaga tatgtgggcc tgatgctgga acttccagtg caatggtggg acagactttt9421tttccagtgg cttcttttct ggtctctctt tctctctctc tctttgagac agagtcttgc9481tctgttgccc aggctggagt gcagtggtgt gatcttggct cactgcaacc tctgtctctt9541gggttcaagt gattctcctg cctcagcctc ccaagtagct gggactgcag gcactcacca9601ccatggccca gctagttttt gtatttttag tagagatggg atttcaccac gttggccagg9661ctggtctcga actecaggac tcaagtgatc tgcctgcctt ggcctcccaa agtgctggga9721ttacaggcat gacccaccgc gcccggcctg gactttcttt taaaagcggc tagagggcca9781ggcgtggtgg ctcaagcctg taatcccagc actttgggag gccgaggtgg gggggtgggg9841gtggggtgga tcacctgagg tcaggagttc gagaccagcc tgacaaacat ggtgaaaccc9901cgtctctact aaaaatacaa aaattagcca ggagtggtgg caggcgcctg taattccagc9961tactcgggag gctgaggcag gataatcgct cgaacccggg aggtggaggt tgcagtgagc10021caagactgtg ccattgaact acagcctggg caacaagagc gaaactcctt ctcaaaaaaa10081aaaaaaaaaa aaaaaagtgg ttagagtact agccaggggg aagcaagcac atttgaaaaa10141tatagtgtgt atgctttagg atagggagga atggagagag agaggaggac atggaccggg10201gtggaaaaat gtggggtttc gcattgattt ttggtgtgag cttgaacaag ttttgtacta10261aaaaaaatgc ctgttttact actaaaataa gtgcttcctt tggagagtga ggatcctctg10321ttactgaggt ttgtattttc aggccctagc atataaatct gctacatacg agatattcaa10381ctggttttgg aggaatgaag gtgaacatgg cagtttcctc atctgtaaaa gcagggtttg10441gacgaaatga tctgcaaaat gggtcttttt gtactcttac tactctagta tttacgagta10501accgatgtta ttccttataa agtctcaact cctggtKtga gccatgtctg acgagggggc10561ctaactccaa taccaccctg aacactctgg ggtcagtggg accctgtttg tacagacgtc10621tgcgtcttcc ggctctttag gacataagca gtactgctat ggaagaaagg aaacctaatt10681tcccatgcct tgtccctttc tggaaagggc atggcgtggt gtgggcacca cacctgcccc10741agccttctag caagaaaaca acacaacctt ccccgctccc cgaaaagagc ttctctccag10801ttgggtctag atgccccctt cccaaccagg gcggatggcc aaaggctccg ctcccttgca10861gggtaggctt ggaaaggcac ggggcccgcc ttcctgggca ccggagcctg tgacgggcgg10921agggcgtgcg gcgcgctggg ttagttacct ggcgctgggc ggggcaagcg ggaggcgggg10981gcggggccgg ggtgacatct caggctgtga ttgggccgcg ccgcgcctgt ctccatgtgc11041cgcgcagctg aggcacaaca ttcaagcccg ggggtggggc cggcgcgcgc cgggaggaag11101cggccgcgcg gcagctgcgg ggcgtggggg tggtggtggc ggcggcggtg gtggcggcgg11161cggcggcggc ggcagctgag gccgaggagg cggtggctgt ggcggacgca gcagcacccc11221agctagggac agggctccgc cgcgccccct tgctggcccc atggaggcgg cagcggcggc11281ggcggcggcg gcagcggcag cggcagcggc gggcgggggc tgtggctccg ggccgccgcc11341gctgctgctg agcgagggcg agcagcagtg ctactccgag ctcttcgcgc gctgtgccgg11401cgccgcgggc gggggccccg ggtctgggcc ccccgaggcc gccagagtcg cccccggcac11461ggccactgcg gccgccggcc ccgtggctga cctgtttcgg gcatcgcagc tgcccgccga11521gacgctgcac caggtgggtc cctccgcctc ctgtccctgc gggtctgtgg ggcagtgtgt11581gcgggggcgg aggttgcgag tggggcgaca gggctgcccc cagggacccg gaggcctcgg11641ccagttctga gagagaggac tctgcagccg gggatggcgg cgacgccctt gtccccgggg11701gacaccagcc tccttcgcag ctcagctcct cgcccagtgc tgggtgcgct ctccccatct11761tccttccctg agacagtctg acactctctc ccctcccccg gccatttctg gggttctgca11821gccgcccaga cccaaagttt gtcctgttct ggaagtggtg tttgttttgg ctgctgcctg11881gtttaacctc tgctcgctgc tcctgcccct ccctctcctg agtgggttct cagggtgcct11941tgctgggatt tcaacttgaa gggtgtgtgt gtccgtaggg ttcgaggccc agggctgaat12001gttgtcctct ctatgtagtt gtaatatata agtatctact atatgtaatt gcccataaca12061tatttctaaa tattaatatt aaatcgaaaa ccttaatgac aagttgcaga acagggacac12121gggactgcaa gaggacgttt gtcaagtcaa aaccaaattt cccatttgag attagaggtg12181gacttcagaa gtccaccatg ggaaacagct gtagtttccc atctatcatt acttggactt12241gaaatagaat tgaaaggtaa gggatgtcat agctaccacc ttttggctgt tctatacttg12301gaaaagtact aggggatgat acataccaga cgctgttttt gttttgtgct ttccttggag12361cttggagaaa gactacttga taagcctttg tctgtcattt catgtcacca gcacttcatg12421gtgataagac tgttaatatc aggttcaaaa cgtgaaatga agcttttcct aaaagcaata12481gtatgaatgt tattaacacc aatggaatat ttagagtggc gagtggtatt ttaaaatcta12541gtttcaaagg agaactgagg gttttatggt tcacttgcaa tgtcattgtg ttcatacgta12601tttttaaaaa aattgcttaa gactgcgttt tatactgctg gagtgttttt actgctatat12661gtttgtgagc ataactttag acactttgta atctttcacc cttttgaatt tgggatttct12721gaaactgtaa ctagtattca cagaacttag catggctgtt tatccctaaa gaagtccttg12781atatgtagac agttctgcca ttgctgatga gtagttgagc tggatcaggt aaagagaaag12841tcagtgtctt aaaacacaaa ccgtggctat tacatgaggc cctcagaata gtcccgagca12901taatacattt attcagtgct gtagctgctt cccagctgaa agaggaataa ttgactgagg12961ctatgcagca agtaagtgac agctgggtac ctattaaggc ttattttgtt gtcagtaatg13021catcggataa gaggaggtgg gcatagtaaa tctgaatatg ttggttgatg cctctttagg13081aattgggaag aaggatttcc tgatgttaga acccatgctt tcttgaggta catatagttt13141taaaaggacc caaggcacag tggctgtgcg tgaatgagat tatatttttg atgtccagtt13201gaaatctgct tgcaaaagac tgcttcagat actggcaaaa ggggacatcc ctgcgagatt13261gcttggagta aatctctcag agctcgaagc tcccttattc agcctagcaa agcaaaacac13321atcatccacc tttaatccaa caaaattacc ataactcaat attcttggta caggtgaaaa13381tgaacattga aggcatcctg gacatgtgat cctaagagag tcaaacaaag tcagcaacca13441gcagacacct aattttcacc catcactttg gtggtggtgg tggtggtggt ggtgNtggtg13501gtgtgtgtat gctccatcca ctttaatttg cattacccca tcagtgtatt agccattctt13561taaaacaaac gtagatgttt ttaccctttc tctgttaaac aggaacctgt aagttgagat13621ttaagtgccc caacaggtat aactgctcaa atcttggagg cactaaccag tcctgatatt13681ttctatggta acaaccaggt tttcctctta ataagcatac aattaagatt accataataa13741tgttccagga agagtgagaa tcttgcattc ttaccacaag taaatggtct ataaaaatac13801ttatttttta tttatttatt ttttgagatg gagtctcgtt ctgttgccca ggctggagtg13861Yggtggcgtg atcttggctc actgcaacct ctgcctcccg ggttcaagcg attcttccgc13921ctcagcctcc tgagtagctg ggactacagg cgcacgccac ctctcccggc taatttttgc13981atttttagta gagacggcgt tttgccatgt tggccaggct ggtctcgaac tcctgacttc14041aggtgatcca cccgcctcag cctcccaaag tgctgggatt acaggcgtga gccaccatgc14101ctggccttaa catatttttt aatagactat tttttttttt cagagcagtt ttaggttcac14161agcaaaattt agcagagtac ccatatatcc cctgccccga cacacccgta gctttcctta14221ctctcaacgt cccacatcag aataatagag tttttacaat tgatgaacct acattgacac14281ctcattattc cccaaagtSt atatattagg gctcactctt ggtgttgtac attctgtggg14341tttggacaaa tgtataatga cggttccacc attgtagtat catacagaat agtttcactg14401ctctaaaaat ctgtgctcta ttgattcatc cctccctctc cactaacatg tggcaatcac14461tgattttttt ttttttttta ctgtccccat agttttgcct tttttagaat gtcatatagt14521tggagtcata cagtatatag gcttttcact ttggcttctt tcatgtagta ctatgcactt14581aaaattcctt catgtctttt tatggtttgg tggctcattt ctttatagtg ctgaataata14641ttccattgtc tgcatgtact acaatttatt tattcaccta ctgaaggata tcttggttgc14701ttccaagttt gggcaattat ggataaagct gttataaaca tctgtatgca gccttttttg14761ttacatgcag gagataagct tttgactcat ttgggtaaat agcaaggggc atgattgcta14821gattgtatgg tgagagtata tttagtttta taagaaactg ccaaactgct ttccaaagtg14881attgtaccat tttgcgttcc catcagcaat gcattggttt ctttacagag tatttatttt14941cctttgctga cttattttaa cagcaggatt ctagcactag gtggaaaacg tatatgagtt15001tctgatgtgt gtcaggtact gtcctaggtg ctgaggttat aatagtgaat ccaacacaat15061ctctgtcttc atggagctta tgttctcatg tgaatgtttt tacacttcag agtttttatt15121tggttgagaa aagacttgaa atataacatc caaaaataaa atgtcagcct ggtgctgtgg15181cgcacgcctg tagtcccagt gctttgggag accaaggatg ggaggattgc ttgaggccag15241gactttgaga tcagcctggg taacatagtg agaccctgtc tctacacaaa ataaaatagc15301cgggcatggt ggcatgcgct tgtagtccta gctactctgg aggctgaggc aggagaattg15361cttgagccca ggagttcaag gttatagtga actatgatcg caccattgct ctccagcgta15421ggcaacagag caaaaccctg tatcaaaata aagtaaaata agcctgtcaa acccaaacca15481aatacatgtc aaacccagac ctgattctta ccacacacta tgccttctag taaaattctg15541gcctttgaat accaacaaaa tacttcataa cctgggaaac caactactct aacgtttgtt15601gctctgtctt gaaatactaa acaaaaccat ttacctgttc gtagacatat tttctttttc15661tagtttaaaa gtatcccatt ctgccatttt ccacctagtg ctagaatcct gctcttaaaa15721taagtcagca cagggaaata aatactctat aaagagacca aaagaccttt caaaataggc15781tatggccagg cccagtggct cacgcctata aatcccagca ctttgggagt ctgaggtcgg15841aggattactt gagcccagga gttctagact agcctgggca agatggcaaa accccatcta15901cacacacaca cacacacaca cacacacaca cacacacaca cacacacaca cacacacacc15961cccgctacct acctctctct ggggcatgcc tgtagtccca gctattctgg aggctgaggt16021gggaggatcc cttgagccca ggagtttgag gctgcaatga gccattatga caccactgca16081ctccagcctg ggtgaaagag ccagaccctg tctcataaaa taaataagta aataaataaa16141atatctacct taaagcagca agttttgtat ttgatgtaat gaattaacag accgctttat16201aatatctatc ttttaggata gtgtcagtct cttggaatgc tgtgcatgag tcatcattaa16261aattggtttt cattaaaaaa atttttttaa catatacaca tcttattgga gttttctttt16321aaaattcaaa aatgggaaca ttattttttc aaagccaagt aacagaagag tatataaaaa16381agtcaatttt gtgcatatct ttttacattt tctatgctca tgtaaaacat acatgactta16441tgtaattata cacatatgct ccagatctca tctgtattaa ttttttaaat tatactttag16501ttctgggata cacttggaga acgtgcagtt ttgttacata ggtatatata tgtgccatgg16561tgatttgctg cacccatcaa cccatcatct acattaggta tttctcctaa tgctatctct16621cccctagccc tgcacccctg aataggcccc ggtgtgtgat gtttccctcc ctgtgtccat16681gttttctcat tgttcagctc ccacttatga gtgagaacat gcggtgtttg gttttctgtt16741cctgtgttag tttgctgaga atgatggttt ccggcttcat ccatgttggt tttcattttt16801gtgatagtta attcttggac attaagggga ataattataa aacactgttg cttccatatc16861aagggaaatc actgaatgat taaaaaaaac ccaataagtg ttgtctgatt gtttaaacag16921ctttcattga ggtaaataag taaaacaaag ctgtgagaag tagggaatat ttaaacaatt16981tgtgcgcatg ccctcacctt tggggagaag tcttttctga aggctctctt ctgatttttt17041gaagcaactg tttaatcatt aaagaaaatt ataggtaaaa gacaaattct tgtgaacttt17101cgctcattgc tagattatta ggcagaacta ttttttaaaa atttaaatga aacaacttta17161tatttttaca tctttagatc atatcatttt gtaactatac tttagggaaa tagaggccag17221catattaaaa cagtaactag gggctgggca gggtggctca ttgcctgtaa tcctagcact17281ttgggaggcc gaggcgggca gatcagctga ggtcaggagt ttgaaaccag cctggccaac17341atagcgaaac cccatctcta ctaaaaatac aaaaattagc aggagaattg cttgaaccgg17401agaggcgaag gttgcagtga gctgagatca ggccattgca ctccagcctg ggagacaaga17461gtgaaactct gtctcaaaaa caaacaaacc aaaaaacaaa cacacacaca cacacacaca17521cacacacaca cacacacaca cacaaacaca caaaccagta actaggacta actatatgct17581gttttagatt gacagtattc tttttttcta aaagagaatg aatgccagca gatgtaggaa17641gtaccagagc taaaccattg cctggttaaa aggagtacct ctgcataata gttacttgtt17701gactcttttg taataaaata tttcaggcat gtaaaactag acagttaccc aagaaataac17761tgtgtatcca ccactccgta taagtaaaac agcccagcta gagtgaaagc ctgctatgtc17821cccttctctg gtcccattcc cctccctccc taccctggag ttaccagagt cctgaatttg17881gtgtgcatga tttgaagatt tctttctact tttattgtat gtatatgtgg atatatatat17941ttgtgtatat ataaatcact aaaaaagata ttattttgga tgtcttttca acttaaggaa18001tatcgtattc tgtgtaacct tctgcaactt gcttttcctc tcattgtttt gtttttggaa18061ttcatcttgt tgacacatgt aatttcagtt catttatgtt tccttttaaa aaatctttac18121tacttacgtt tacaatgact cattgttaca gtgcggtaga gacccttatg ccaaaaggaa18181gtaggtggca tttttcctta tttatataga atgtcacatg actgggggac tgttcgaaac18241gtgtatgaaa attaagaaca ccttcataga tagcagaatg cacacttata ttcttgagcg18301tgctagggga tgtttgagct aattaattga ctttcatggg ctttcttttt ttctcatctt18361ttattttttt gagacagggt cttgctgtgt cacccagact ggaatgcagc aatctgatca18421tgactcactg taccctcaac ctcctgggct caagcaatct tcctgcctca ggctcccgag18481tagctgggaa tacaggtgtg cagcaccaag cctggctaat tttcttt-tta aaaatttgtt18541gtggagatgg tgtctcatta tgttgcccag cctggtctca aactcttggg ctcaagtgat18601ctcccgcctc agcctcccaa agtgttggga ttacaggtgt gagccaccac accctgccat18661ttttcatctt tttaaaaatg gaacatctca aataagcaag tagatagaat aatgcattaa18721accctgctgc acttgccact caggcccaac aactgtcaat gcaaggttaa ttcaattcca18781tctacatttc tatctttgtt ccctctcacc tttgttatcc tgaagcaagt cgcaaacatc18841atatggttca tagggcttat ctttaaggaa ggaaccaggg tgggcagaag gaaatggaaa18901catgcagggg tttaggttgg ccagaaaagg tacctgtggc ctatggtcaa atggcaagac18961tgatgatggt tcaatactgt caagcctttg gctgtcatgg agatagacgt gcttgagaac19021tgatttagaa tagttgggaa acctgtaggg agattgaaaa agtaaccaac tgaagaccag19081gaatacaaga tggatttgaa tattgttcta gcttcacact cctaagtagg ggctgtccta19141ttttaagggt gtagacaatt ggttttgagg aaatcttggg ttgtttttgc taattattta19201taactttttt tttttttttt tttttttgag acagagtctt gctctgtcgc ccgggctgaa19261gtgcagtggc gcaaggtcgg ctcactgcaa cctccgcctc ccaggttcaa gcagttctcc19321ctgcctcagc cttctgagta gctgggacta caggcgccca ccaccatgcc tggctaattt19381ttatattttt tagtagagat ggggtttcgc catgttgact aggctggtct tgaactcctg19441acctcaggtg atctgcccac cctggcctcc caaagtgctg ggattacagg tgtgagccac19501cgcgcctggc ctatttataa ctttaaaaag agggtttggt tcctttccag ggtttttcaa19561actttctgtt tgtttcgtat aggcaagttt ctcatgtttt gtattgatgg cattctatta19621ttcataccta tctgttggtg ggcaactaat caacctgggt tgcgcaggac tgaaggattt19681cctgggggca tggaacttca gtcttcagac agtcctaggc aaactgggat ggtcaccccc19741tctcttggtt gctgcattag ttatctattg ctctataatg aattaccaca aacttagcag19801cttaaagcag cacacagtta ttatctccca gtttctgtgg gttcgggagt ctggacatgg19861ctcagttggg tcctcagctc agagtttcat aaagccacaa tcaaggtgtc tggagcttgg19921agtcctctgt ggaggtccaa cacttggttg cagaattcag ttccttgtga ttttagaact19981gaggctctca acttcgaggc ttcctgcagt tccctccaca tggccttctc cataggcaat20041tcacaacatg gctcttggct tcttcaaggc cagcaggaga gtgtctctct ccagttagct20101cagacacctt tgccatagtc cattagttag aagcaagtca caggttttgt ctgctctcaa20161ggggagggga ttatacaaaa caacagagac caggaggtga gaattttgcg ggggacacct20221taggatcttt ccaccacact tgctttccag gatttgtctg aagtagaaaa ttattctgtt20281gagccttttt gtggccagaa tatacatatg ttttgctcag ccactaattt agaattcctt20341tatcttttta agaaacattt tcatgcgtaa ttcatcctaa gtagaactgg caaatatgta20401acttcttagg tattttcaat tttagtcttt aggcaattgt ttttcctctt aactagtagg20461aggcaggaaa aaaaagtgga aaagcagatg caaatgccct gcctggctac tcctgcacgc20521agtgctctaa agcaggcctt ctcagcctgg gttagttcag gggtctgtga atttagatgt20581ggaaaagaac tacatctttt tttttttttt tactaactgc ccttccccca actgaaattg20641agcacttcct tctattatgc atgtaagcaa aaaaccacag cagttttttt tttttttttt20701tttttttttt ttttttttga gacagggtct tgctctgtca cccaggctgg agtgcaatct20761cacaattttg gttcactgca acctgtgcct cctgggccca aatgatcctt ccatctcagc20821ctcccaagta gctgggacta cacgtgcatg ccaccacacc cagctaattt ttgtattttt20881tgtagaaacg ggattttgcc atgttgccta ggcaacaggg ttcactgctt tcggtcaaat20941gacagcaatg ctgctggcaa gtccatggtc aggagtgaga ttccctcagg ctgctgtggt21001ccctcctcct ccagtctgtc ctgcaggaag ctcagagtga atggatgctc agctgtttcc21061tctgagacct caccagggac agttcctgca gtcccggctc tgagctctgc tcctgaccgt21121tttatctggc cctctcagct gtgcttcact gcaaaggagg aaggacaagg ccctctggag21181ccagtgccag ctgcttgtgg ggcctgtttt atttactact gctatctcta cttcagagac21241ttgggctagg acatagactt gaagaaaaaa gccattttaa cagacttgag aaactcagcc21301atcctcataa taccattcag aaccaggaga ggccattatt aacactaggc tagtaggaaa21361tgttcacttt gactgaattc atttgtaaca ctttgccact ggtttacaag aaaaatactt21421tggttcagtc tcctggaggt aggtaattaa atgtttcacc tgtgctaatt ttccaaataa21481ttaaagtgaa tcccataaat gccagcaatt aagaattgtt gtttttggcc aagtgtggtg21541gctcatgcct gtaattacag cactttggga ggttgaggcg ggcggatcac ttgaggccat21601gagtttgagg ccaacctggg taaaataggg agacgctgtc tctacaaaaa atacaaaaat21661cagctgggct tggtggaatg tgcctgtagt cctggctact tggtaggctg aggcaggaga21721attgcttgaa tccgggaggt agaggtttca gtgagccaag attgtgccac tgtactccag21781cctaggcgac agaggcgaga ttctgtctca aaaaaaaaaa aaagttgttt cttaagtgca21841gaccttctga gatgtgtagt gcttgcccca tgccttcaaa agaacatatt taatacaata21901atctctttgt ctctccttta cttcaaagtc agccttgtga aaaacagatg atgatatacc21961aacagttttt tgctcttgga ggtgtttggt gtgctctgct ttttgagttt tgttttttgg22021acttctttaa aattatttta aaatgtttta actcctctcc tttgcatact ctctggtacc22081cttatagtcc caatactgag taaagcatgt aaaatttagc atgcagatgt tttagcctct22141tttatttttt gttcgagatc cagatttttt agtctcagca tgactgacac tttggggctg22201tatcattctt tgttgtggag gactgtcctg tgcattgtag gttgtcaagt aacatccctg22261gtctctaccc acaagatacc agtagctatt cttctcctga ttgtgacaac aaaaaatgtt22321tccagacatt gtcagatgtc tcctcggggc aaaatttagt tctacatata tgatgataaa22381aaccatggta gcagccaaca tttgttcact atcatcccca ttttgaagaa agcaaaaacg22441gaggcactga gggtgaggtt cacttacttg cccaaggtca catagataaa agtcacttct22501tgaaatactt ttcataatat atatgggttg aatctcagtc ttaattgagg aggcagacta22561aaaaagaatg taaaaacggg gagtttcctg atgatgtctc aattgagagg ctattgcttt22621catagtgatt ctgtgtaaat gctggaattc ttgcttaaac ggggtgattc catgggcatg22681tgtctgagta aaggacttgg atcatttcta agaagcttct gtggcatggt gaagcagaac22741ataagacagt ctaggatcaa ggataactga tcaaactttg aaaccaagga agccatagga22801aattggggta aggctatgta tcacttaggg tcccacagga aatggcatgc tcaaagggtt22861ttagtgaaaa caggaatgtt gaggcacgaa gggactagca gcaggaagct attgccattc22921ctagacctgg tggggcaaag ggaggggagg gaatgtcgtt gctggagctg tgaaaggtac22981tccctctaca ggaacaatag tcatgaaaga atacagcctt tgctagaact atggccagag23041aggggccagt gggatgaggg gatgggagag aagtaaccaa tttctgtttc ctgccctctc23101atttgctgca gtgccttttg ttgttctaac ccaactagaa gccacaggta acagagcctg23161ggtgttgaag cttattgagg ttagcctctt gcgctgagtg gatggaaagg gctgagagtg23221gttcaagggg gcaaatagag caacagccag cacaggctat gagcaggagt gggaaacact23281accctggatc ctaggaagga tggggcatgg ccagtgatag gagcaatgaa agtctttttg23341tgtgaaaaag agcctgggga tacagacctc catgaaggag aagagagggt tggataggag23401tgactggttc atctgaggtt cttcctattt ctccaaggaa gccttgagga atgtggggtt23461gtctgtttgc atttggatat ggaagtatgt gtgtgtgtat acttttattt attttgagac23521agggtcttgt tctgtcaccc aggctggagc acagtggcac catcacagct cactgcagcc23581tctacttcct gggcttaagt ggtcctccca cccaggcccc cccaggtagc tgggaccaca23641ggcatgtacc actatgctcg gctgattttt aatattttgt agagacaggg tctcactata23701ttcccctggt tggcctcact cctgggctca agtgatcctc ccaccttggc ctccagaagt23761gctaggaata caggtgtaag ccactacacc cagtctgttt gtatactttt tggtcagagt23821gaggctttcc gtgagttgga gtgtttgtcc ttattcatga cactgggaaa catacaacag23881tggagtgacc ctcttgcaga ttctgctagg agtagctaca tatggtggga aaggttgcta23941tgggaatgga gataaattca aRgcagtaga gagtgcttta taatgtttgg gggtatggcc24001aagcattagt tttgcttttt ccttcccagt aggatacgat ttctgtagca attcttcctt24061acccaaaaat agctatatgt gctcccaaaa agaaaaagcc caggaccaga tgacttcatg24121gctgaattct accaaacatt caaagaagag ttaatactaa tccttcttaa actcttccaa24181aaaacagaag tagagagagt acttccaaac tcattttttg aggccagcat caccctgatt24241tcaaagccag acaaaagata ccaagatacc ataagaggag aaaaccacag gttagtatct24301ctgttgaaca tatatgcaaa aatcctaaat aaaatactag cagaccaaat ttaacaacac24361ctcaaaaaga tgatacacca tgaccaaagt ggcacttatc cctgagatgc aagattggtt24421taacgtatat aaaacaatca gtgtaatatg ccacattacc aaaataagag gtaaaaaaca24481agccaggtgt ggtggctcac acctataatc ccaacacttt gggaagctga ggtgggagga24541ttgcttgagg ccaggagttc aagaccagcc tgggcaacat agtgagacac tgtctctaca24601aaaaattaaa aaaaaatgag ctgggcatag tggcatgtac ctctagtccc tgctacttgg24661gaggctgagg cagcaggatc gctggagccg aggattttga ggctgcagtg agcactgatt24721gtgccaccgc actccagttg ggcagtacag caagaccctg tcttcaaaga aaaaaaaaga24781gagaaaaacc catgatcatt tcaatacatg tagaaatagc atttgacaaa gttcaacagc24841ctttcgtgat aaacattttc aacaaaataa gtttaaaagg gaatttcctc aacacattaa24901agaccattta tgtagagtcc acagctaata tcataatcag tgggagaaaa ctgaaagctt24961tttttctaag atctggtaca aagcaaggat gcccactctt gccactttta ttgaacacag25021cactggaagt actaacatga gctatcagac aataaaaaga gataaaaggc atctaaattg25081ggaaagaaga agtaaaatta tccctgtttg taaatgtcat gatcctatat gtagaaaacc25141ctggactcca caaaagccct cttaagacta ataaatgagt tcagtaaagt tgcatgatac25201aaaatcagcg tacaaMaatc agttggaaat caagaaaact gtcccattta tgataacata25261aaaataataa aatattcagg aatacattta atgaaggaag tgaaagatct gtacattcaa25321cattataaaa tattgatgaa agaaattgaa gacacaaata aatggaaaga tatgccatgt25381ttatgcattg gaaaaattaa tagtgttaaa ttgtccacac tacccaaaac aatatacagg25441ttcaatgcaa tccctatcaa aataccaatg gcatttttca cagaaataga aaaaacaatt25501ctaaaattca tatggtgtga caaaagacct agaatagccg aaacagtctt gagaaacaaa25561aacaaggttg gaggcatcac acttccttag attataatac agtataaagc agtagaaatt25621atgatagtat atcctggcat aaaaacagac atatagatca atagaataga acagagatcc25681gagaaatata cagtatatat acagaaatcc aagtatatac agtcaactaa tttttgacaa25741ggccaccaag gagacacagt ggggaaagga tagtctcttc aataaatgat attgggaaaa25801ctggatattc acaggcaaaa gaatgaagct ggatgcttat atacaaaaat caactcaaaa25861tgtgtcttta aaatatacaa caaaacctga aaatggaaaa gaagacctaa atgttaataa25921ctcaaaccat tgaactccta gaagaaaaca taggggaaaa gctccttgat atcggccttg25981gcaatgattt tttagacatc acaccaaaag cacaggcaac aaaagcaaaa agaaacaggt26041ggtactacat cagactaaaa agcttctgca tgacaaaggg aacaatccgc aaataaaagg26101gcagcctatg aattggaaga aaatatttgc gacccatata tatgataagg agttaatatc26161aaaaatatat aaggaactcc cagaactcaa tggtaagaaa atgaataacc cgattaaaaa26221ataggcaaag gacttgatta ttcattttcc caaagaagcc atacaaatgg ccagcaggtt26281gtgaaaaggt actcagtatc actaatcata agggaaatgg aaatcaaaac cactatgaga26341tatcagctca caccttttag aatggctatt agaaagacaa cagataacaa acgttagggt26401gtggagaaaa gagaaccctt ttatactttt gatgggaatg tagattggta cagccattat26461ggaaaatggt atagaagtgc ctcaaaaaat taaaagtaga actaccatat gatccagcaa26521ttcctctgct ggatatgtac ctaaaggaaa tgaagtcagt gtgtcaaaga ggtatctaca26581ctcctgtgtt cattgcagca ttattcgcaa tagccaagat atggaaacaa tttaagtgac26641tgttgatgga tgagtggcta aaaatatcgt gggatacaca cacacacaca cacacacaca26701cacacacaca cacacacaca cacagatgac agaatattat tcagccttaa aaaaggagat26761catgctattt gtgacaacat ggatgaactt atgctaagta aaataagtga gacacagaag26821gaaaaatact tcatgatctt atttatctgg aacctgaaaa aaagagtcaa atatatagaa26881acagaataga accacggtta ccaagggtgg gaatggggag gaaataaggc aatgtaggtc26941aaagtgtaca aacttgcagt gatgtaggct caataagtct agagatgtac agcaggagga27001ctatagttaa caatattgta ttgtatatgg aaaatgtgct aagagtagat tttaggtgct27061cttacctctt ctccccacaa aggtaactat ggaaggtgat gagtatgtca gattgcttac27121tggtagtaat catttcacta tggatatgta tatcaaaaca tcatgtcata catgttaaag27181ttatgccata aaaatagcta tatgtgggcc aggccaggca tggtggctta tgcctgtaat27241accagcattt tgggaggctg aggcaggagg attgcttgag gccaggagtt taagaccagc27301ctgggcaacg tagtgagaaa cctgtctctc caaaaatgtt tttaaaagtt atctgcgtgt27361ggtagtgcat gcctgtgatc ccagctactt gggaggctga ggcaagagga ttgcttgagc27421ccgaaaggta gaggctgcag tgacctgcga ttgcaccact gcactccagc ctgcgtgaca27481gagtgagacc ctgtctctta aaaaaaaaaa agattaaaaa atagttgtat gtgtatatgt27541atacgttcag gatttgttct atttgttttc tttttctttt ttttcatagg cagtggtcaa27601tggcctaagg atttctttat acattgctca tatccactaa ggctctattg agcaaaccac27661gccttaaaaa tcttacaatc ctcttcatat attttaaagg caaatgcaaa gaaatagctt27721tgttctgttt tgcattcttc atgggttaac atatcttctg gaaaacttgt tgctatggca27781atgggatatt atctatcttg gtaatgtact acataaaaat agccctttgt gtttgagaat27841gatgttagtg acaggtggtt gggtgaatgt atagcaggct gctttgtcaa gaatagtttc27901ccaacattcc attgtttaaa gttgtgaaat caaaatgagt ataatttttt tctctctcaa27961aatgtaattt tatctggaaa aggcaggcag tggtgctctg tttaattatg aattacaagc28021cttcctttgc aggtcaggtc aacatgatga ttgaagccac ctctcactgt aataagtaaa28081cgacttctct attctttctg tcagggtgtg cagcacaagg ttgcgctggg ggggccagta28141gggataattt tcccattctt gaacacatcg ttcttcctaa gcacctcgta cttataaacc28201aagaagtagg aagtggtcag gtggaagaga tgagctctga gctgttaatc aaatccgaat28261ttcttgtagg ttcttaggtg aaacagttcc ttgggaaagg tcagatttcc atgttgcttt28321tagtattgaa ctcacacata tagccaagga tagaatctga cccaaatctt aatttttcct28381ttgaaaaatc ctttgggcaa ttttttttta atttaattta attttattat tattattatt28441attttttatt gatcattctt gggtgtttct cgcagagggg gatttggcag ggtcacagga28501cagtagtgga gggaaggtca gcagataaac aagtgaacaa aggtctctgg ttttcctagg28561cagaggaccc tgccgccttc cgcagtgttt gtgtccctgg gtacttgaga ttagggagtg28621gtgctgactc ttaaggagca tgctgccttc aagcatctgt ttaacaaagc acatcttgca28681ccgcccttaa tccattcaac cctgagtgga tacagcacat gtttcagaga gcaccgggtt28741gggggtaagg tcaccgatca acaggatccc aaggcagaag aatttttctt agtacagaac28801aaaatgaaaa gtctcccatg tctacctctt tccacacagt cacggcaacc atccgatttc28861tcaatctttt ccccaccttt cccccctttc tattctacaa aaccgccatt gtcatcatgg28921cccgttctca atgagctgtt gggtacacct cccagacggg gtggtggccg ggcagagggg28981ctcctcactt cccagtaggt gccgccgggc agaggcgccc ctcacctccc ggacggggcg29041gctggccgga cggggggctg acccccccca cctccctcct ggacggggcg gctggccggg29101cagaggggct cctcacttcc cagtaggggc ggccgggcag aggcgcccct cacctcccgg29161acggggcggc tggccgggcg gggggctgac ccccccacct ccctcccgga cggggcggct29221ggccgggcag agtggctcct cacttcccag taggggcagc cgggcagagg cgcccctcac29281ttcccggacg gggcggctgg ccaggtgggg ggctgacccc gccacctccc tcccggacgg29341ggcggctggc cgggcagagt ggctcctcac ttcccagtag gggcgcccgg gcagaggcgc29401ccctcacctc ccggacgggg cggctggccg ggcggggggc tgaccccccc acctccctcc29461cagatggggc agctggccgg gcggggggct gaccccccca cctccctccc ggacggggcg29521gctggccggg tggggggctg acccccccac ctccctcccg gacagggcgg ctggcagggc29581agagtggctc ctcacttccc agtaggggcg gctgggcaga ggcgcccctc acttcccgga29641cggggcggct ggccgggcgg ggggctgatc cccccacctc cctcccagac ggggcggctg29701gccgggcggg gggctgaccc ccccacctcc ctcccggacg gggcggctgg ccgggcgggg29761ggctgacccc cccacctccc tcccggacga ggtggctgcc gggcggagac gctcctcact29821tcccagacgg ggtggctgct gggcggaggg gctcctcact tctcagacgg ggcagctgcc29881gggcggaggg gctcctcact tctcagacgg ggcggttgcc aggcagaggg tctcctcact29941tctcagacgg ggcgtccggg cagagacgct cctcacatcc cagacggggt ggcagggcag30001aggtgctccc cacatctcag acgatgggcg gccgggcaga gacgctcctc acttcccaga30061tgtgatggcg gccgggaaga ggcgctcctc acttcctaga tgggatggcg gctgggcaga30121ggctgcaatc tctgcacttt gggaggccaa ggcaggctgc tgggaggtgg aggttgtagt30181gagctgagat cacgccactg cactccagcc tgggcaccat tgagcactga gtgaacgaga30241ctccgtctgc aatcccggca cctcgggagg ccgaggctgg cgggtcactc gcggttagga30301gctggagacc aacccggcca acacagcgaa tccccatctc caccaaaaaa atacgaaaac30361cagtcaggcg tggcggcgcg cgcctgcaat tgcaggcact cggcaagctg aggcaggaga30421atcaggcagg gaggttgcag tgagccgaga tggcagcagt actgtccagc ttcggctcgg30481catcagaggg agactgtgga aagagaggga gagggagacc gtggggagag ggagagggct30541gggcaatttc ttagagcagt aactctcaaa cattttgtcg tcatcatctc acttttcttc30601tacttatcaa aatgagtatt gatttaattg taatgagtaa ataacagctc cctgctactg30661aaatgaaatt tactctagct ttgagaattt gagagaccct cttaagaaaa taccacagct30721ggaaatttga gacctaaaag taatacaatt tgagtatctc atatctgaaa tgcttgggac30781cagaagtgtt ttggattttg gattttttca gactttggaa tatttgcgtt tacataatga30841gatctcttgg ggatgggacc caagtctaaa cacgaaattc attaatgttt tatatacgcc30901ttatacacat agcctaaagg taattttata caatatttta aataagcttg tgcatgaaac30961aaaattttga ctgtgttttg actgtgacct gtcacataag gtcaggtgtg gattttttca31021tttgtgactt catgtcaaca ctcagaaact ttcacatttt ggagcatttt ggatttcaga31081ttagggatgc ttaatctgca gctctaattc tgatatgcac ggtactactg cagagatgga31141accattgcct ttgtcttttc tgttggagtc agtatagata tcttgctact aataatatca31201gaagagagtg ttgagtggtt tgctgggtgc ccaatactac ttttagaaca ttttatgtat31261taactaattt attcctgtaa acattcttga cagtttttgt gactatttcc caaatgaggc31321gtacttgtta gcatgtcttt ctgtttgtgg tgagtaagca ttcttgtgta ttggaagact31381tgcttactag catgtcacag tgttcttcat attgaaagca ttcttctatc aggagttgac31441gtaattcagc ctgtgggcca aatccatcct gccacccatt ttttgtgtat tttataagct31501aagaatggtt tctgcatttt taaatggttg ggaaaacata aaaagaataa tttgtgacat31561gtgaaaatta catgaaattt aaatttcagt ttccataaac agttttattg gaacacagct31621ttactcattt gtttacatat tgctaaggct gcttttgaac tagagtggca gagttgagta31681gttgtaatag agaccatata gctcccagac cctaagaaaa agtttgccaa tcccttctct31741atattcttct gtatctagaa tggcagatgg tcctcaataa taacagcaga ttggtcaggt31801gtcatagttt atgcctgtaa tcccagcatt ttgagaggcc aaggtgggag gatcacttga31861gctggagttc aagaccagcc tgggcaacac agtgagacct agctctgcaa aaattagcca31921ggtgtcatgg tgcacgctgg tggtcccagc tacttgggag gctgaagtgg gaggattgct31981tgagcctggg aggtcaaggc tggagggaga cgtgatcaca ccattgtgct ccagcctggg32041tgacagagcg agaccctgtc tcaaaaatat ataaacagat tggatttatg tacaggttca32101tatatatatt aattttcttt tggtatgcaa tatatttttc tttcttccac taaacgtttc32161tcttgtcata agcttgatgt catgtactat ggcatatctc aaatatacgt gatttcagct32221ctttgcatac ccatattggt tttctttttt ttttttttaa ttgatcattc ttaggtgttt32281ctcgcagagg gggatttggc agggtcacag gacaatagtg gagggaaggt cagcagataa32341acaagtgaac aaaggtctct ggttttccta ggcagaggac tctgcggcct tccgcagtgt32401ttgtgtccct gggtacttga gattagggag cggtgatgac tcttaaggag catgctgcct32461tcaagcatct gtttaacaaa gcacatcttg caccgccctt aatccattca accctgagtg32521gacacageac atgtttcaga gagcaccggg ttgggggtaa ggtcacagat caacaggatc32581ccaaggcaga agaatttttc ttagtacaga acaaaatgaa aagtctccca tgtctacctc32641tttctacaca gacacggcaa ccatccgatt tctcaatctt ttccccacct ttcccccctt32701tctattccac aaaaccgcca ttgtcatcat ggcccgttct cagtgagctg ttgggtacac32761ctcccagacg gggtggtggc cgggcagagg ggctcctcac ttcccagtag gggcggccgg32821gcagaggcgc ccctcacctc ccggacgggg cggctggccg ggcggggggc tgacaccccc32881acctccctcc cggacggggc ggctggccgg gcggggggct gaccccccca cctccctccg32941ggacggggtg gctggctggg cagaggggct cctcacttcc cagtaggggc ggccaggcag33001aggcgcccct cacctcctgg atggggcggc tggccgggcg gggggctgac ccccccacct33061ccctcccaga cggggcggct ggccgggcgg ggggctgacc ccccccacct ccctcccgga33121cggggcggcc ggccgggcag aggggctcct cacttcccag taggggcggc cgggcagagg33181cgcccctcac ctcccggatg gggtggctgg ccgggcgggg ggctgacccc cccacctccc33241tcccggacgg ggtggctgct gggcggagac gttcctcact tcccagacgg gatggctgcc33301gggYggaggg gctcctcact tctcagacgg ggcggctgcc gggcggaggg gcccctcact33361tctcagacgg ggcggctgct gggctgaggg gctcctcact tttcagacgg ggcggttgcc33421aggcagaggg tctcctcact tctcagacgg ggcggctggg cagagacgct cctcacatcc33481cggatggggc ggcagggcag aggcactccc cacatctcag acgatgggcg gccgggcaga33541gacgctcctc acttcctaga tgggatggcg gccgggaaga ggcgctcctc acttcctaga33601tgggatggcg gccaggcaga gacgctcctc actttccaga ctgggcagcc aggcagaggg33661gctcctcaca tcccagacga tgggtggcca ggcagagaca ctcctcactt tccagacggg33721gtggcggccg ggcagaggct gcaatctcgg cactttggga ggccaaggca ggctgctggg33781aggtggaggt tgtagcgagc cgagatcacg ccactgcact ccagcctggg caccattgag33841cactgagtga acgagactcc gtctgcaatc ccggcacctc gggaggccga ggctggcgga33901tcactcccgg ttaggagctg gagaccagcc cggccaacac agcgaaaccc cgtctccacc33961aaaaaaatac gaaaaccagt caggcgtggc ggcgcgcgcc tgcaattgca ggcactcggc34021aggctgaggc aggagaatca ggcagggagg ttgcagtgag ccaagatggc agcagtacag34081tccagactcc cacggcatca gagggagacc gtggaaagag agggagaggg agaccgtggg34141gagagggaga ccgagaggga gagggagaga gggagaggga gagagggaga gggagagagg34201gagagggaga gatttggttt tcataataat cataaccctt tccaaatgag ggagagtaat34261gaattttatc attttattag ttagtgtgct tcctttgcta gatcaggaat tatatctcct34321agtaattata accctctatt tattattttt aactatattg ctttctactt ttgggctttc34381agccaacgtg taaatattga cagtttaata atctgaacca agccacctgg actgattgta34441atattttaat ttaataaatc ttgcatatgg ttaagattac atcttctaca ttcatcagat34501tggatttaat gcactcacca taatcaactc tggaacattc ttgcaattat tattgttatt34561tttaaaggtt ttctcctttg tctgttgttt tcccactttt tttttttttt tttaagacgg34621aatctccctc tgtcacccag gctggagtgc agtggcgtga tctcggctca ctgcaacctc34681cgccccctgg gttcaagcga ttctcctgtc tcagcctcct gagtagctgg gactacaggc34741gtgtgccacc acgcccagct aattttttgt atttttagta gagatggggt ttcaccatgt34801tggccaggct ggtctcaaac tcctgacctc aagcagtctg cctgcctcgg cctcctgaag34861tgctgggatt ataggcgtga gccactgtgc ctggcctgtt ttcccacttt ttaaagtaac34921agccttgttg agatgtaatt cacataccat atagtcacct atttaaagtg tacaattcag34981tggttttagt atattcacag aattgtgcaa ccatcacaac tatttccgga atatttttgt35041cacccaaaaa agaaacccca tactcactag caatcacccc cagttccacc attcctaggc35101agccatggat ctactttttg tctatggatt cgtctattcc agacatttca tagaaatgaa35161accacatagt gtgtggtcat ttgtgattgt cttctttctc ttagcgtgtt ttaaatgtag35221cacgtatcag cacttcttat tgccagttct gctgtatggc tataccacat tttatttatc35281aattcatgag ttgacagaca tttgggtcat tttcactttt tgcctcttat gtataatgct35341actctgaaca cttgtgtata ggtttttgtg tggacatatg tttttatttc tgtcgaatat35401atacctagag gtggaattgt tgtgtcgtat ggtaactcta tgtttaacct tttgaggaac35461tgccaggctc ttttgcaaag ctgctgtact atttgacatt cttgtgagca atatatgagg35521gttccagttt ccccacatcc ttaccaacgt ttgttattgt ctatcttttt gattatggct35581atcctagtgg gtgtataatg gtatctcatt gtggttttag ttggaatttc cctgatgact35641aataatgttg catgtctttt cacgtgctta ttggccactt gtatatcttc tttagagaaa35701tgtgtattta gatcgtttgt ccattttaaa ttgggttgtc tttataaaat gttgaattgt35761aagagttctt tatatattct ggattcaata catatgatac aatacatata atacatattt35821tagtcatcta tcagataaat gatttgcaaa aattttcttc cattttgtgg gttgtctttt35881tactttcttg atggtgtcct ttgaagcatg aaagctttta attttgatgg agtttattta35941ttgttttcta ttgtcacttg tgtttttggt gtcatattta agaaaccatt gcctaatcca36001agatcacaaa gatttcactt atgttttctt ctaagagttt tgtagtttta gctcctatat36061gtcagtcttt gattcatttt gagttaattt ttgtacgtgg tgtgaagtag gggtacaact36121tcatactttt gtatatggat atccacttgt cttagcatca tgttttgaac aaactgttct36181tttcctaatg aattgttttg gaacccttgt gcaaaaccaa ttgaccttat gtaaatgtaa36241gggcttatgt ctagactgtg tcagtatcac actgtcttga tttgtttaat ctatttttga36301ggcagacttt cgctccctga tctaaaatta aaatgtttca caattgatga tgtcttatat36361gcatcaaaac ttgcagactg ggtgtaagtg gcttgagaaa atatattgga gactaaagtg36421gggctgttgg agactcatag gaatcaaata gttagaaggt gggggagtga gagaggcagt36481aagttggatg cttcccaggt gtttagcagc catcccaaag tggcagtcaa aaataggcta36541gctttacata atttcaaagt taggtactga tggttccagt ttctttttat ggattctttt36601aaaaagtgct gtatcaccaa atgctgttga tggcatagaa gacaacactg tggaaaaatg36661tggacattga ccagtctgag tcaaaaagat ggacagatat tcagttctaa atgtgaggaa36721tttttaggaa aaccttgagc agttaatttt gcttatttcc ctttctatgt gtatatgaat36781gataaatgaa aagtctaaaa gagctccttt aataagcctg agataaatgc taagtcgtag36841actactgcat catatcttat ttggcttttt tttagtgcca cctaaaatac tggtgtttta36901gctttgagat gctcttatcc tcctgcttcc agctctatct acacctcaga aggagctggg36961tcctttccca gagtctattg tataaattaa ccttcttcct gtagtcactc tatgtccatc37021tgtttcccag ccactttaat tttgttcttt ttcctttctc ttgtgcttgt agatttaagt37081cttaaaaaca agccaggtat tattgtttat agtggaattt tgagtaggag tggaggttaa37141cctgtacatc aatctagtag gaatattcat tcaatcactc aagggatgag cttatggaaa37201agttacaaaa atagtagcat tcctttatat tcctcaccct gctcccttaa tgttaacatt37261ttatataaag gatcaagaat aggaaattag tcttggtaca gtattattaa ctaaactaca37321gagcttattt gaattccatc agtttttcca ctggtggtct atttctgttc taggatcctg37381tccaggatcc catattgcat ttagttatta tttctcctta gtctcatcca gtttgtaata37441atttcttaat ctttaatgac acctttgaag agtacagatc aatattttgt agaatgttcc37501tcaatttgta tttgtttgat gctttcttat gaataaggtt gttcattttt ggcaagaata37561taatgatgca tatttttcag tgtatcatat caaggagttc atgatgttga tgtgtcttgt37621tactggtgac atttaccttg attccttgtt taaattgctg tctgcagggt ttctctacag37681taaaattatt actttttctc tttatggtta aaatatcttg ggggagatgc tttgagacta37741ggcagatatc ctatttatcc tcaaattttt gtccattgat ttttttcatt ctttggtgga37801tcactctgta tcatagagta tttgtgacaa atatttgatt agcagaaata aagctatgcc37861ctacaagaag caaagatcaa ttacattgaa atcctattgt attggttaat ataacgctag37921cagctgtgtt cctcaacttt caggagctta acacaacaga tgcttttctt ctcaatatgg37981gcatttctgg ttgtgtaggc agttttctgc catgttaatt caagtatcca gactaaactt38041tttatactga agtaacttca aactgacaga aaatagggat gcaagaatag taccaggaaa38101tcctgtatac cctttactag atttacttaa catttgctac atgtgcctta tgattttctt38161tgtctattca tatatgtata tgtattttat atatatttta tatatctgta tttatatatt38221gattgccata tactaaatca cttgagatta aatcacatac atcatgttct tctatgcctt38281aatagtttaa acttaatata tttatttgtt taaatctagt atacagtttc agaattgcta38341acccatatct gtttgagaaa cactttcact aactggatta cagcatttat acaattcttt38401tattctttag ttttgcaata tctagtcaag atacgatttt ccagtttccc actgaagtat38461taagtttgaa aactgtacat aagcacactg ttattcaaat ttaggaaatt taatatcagt38521actttagtat gcagtctgta ttccagtttc attaattatc cccaaaatgt tcttcagtgg38581aatatttttt ttctcctgat acagtatcaa gtccaggatc atgtattgaa tttatttgtc38641atgacttttt agtttccttt gatctggaac agatcttata agcctttctt tgtctttcat38701gacatcgaca ttttgaacaa tgacaaattt atgacattgt tttgtagaat gtaccaaatt38761tgggctgggt gctgtggctc atgcctacaa tcctagcact ttgggaggct gaggcaggtg38821gattgtctga gttcaggaac tcgagaccag cttggccaac atggtgaaac cccatctcta38881ctaaaaatac aaaaaacaaa aaaaattagc catggcacct ataatcccag ctactcagga38941ggctgaggca agagaatcgc ttgaacccgg gaggcagagg ttgcagtgag ccgagattgc39001accactgcac tccagcctgg gcgacagagc tagatgctcc atttcaaaaa aaaaaggaac39061gtaccaaatt tgggtttggt tgatggctcc tcttgtttag attgagatta tgcattttta39121aaaaaacttt taggttcagg ggtacatgtg ctgatttgtt gtataggtaa attcttgtca39181cgggggtttg ttgtacagat tatttcatta cccaggtact aaacctagta tgcagtagtt39241ttttttttct cttctccctc ctgccaccct ccatcctcca ataggcctca gtgtctgttg39301ttcttctctg tgtgtccatg tgttctcatc acttagctcc cacttacaag tgagaacatg39361cagtatttgg ttttccgttc ctgcattagt ttgcttagga taatggcccc cagctccatc39421catgttcctg caaaggacat gagctcattc ttttttatgg ttgtgtagta ttccatggtg39481tatatgtacc acattttctt tatccagtct atcattgatg ggcatttagg ttgattttat39541gtttttgcta ttgtgaatag tgctgcaatg aacatacgct tgcatgtgtc tttatggtag39601aatgatttat attcctttgg gtctataccc agtaatgggg ttgctgggtc gaatggtatt39661tccgttttta ggtctttgag gaatcgccac tctgttttcc acaatagttg aattaattta39721cactcctacc agcagtatgt aagcattcct ttttctctgc aacctcactg gcacctgttt39781ctttttgact ttttagtaat agccattctt actgatgtga gatgttatct cattgtggtt39841ttgatttgca tttctctaat gatcagtgat gttgtggttt ttttttcatc tgctcgttag39901ccacgtgtat gtcttctttt gaaaagtgtc tgagattatg catttttgtc tggaatactg39961cataagtgac atgtccttct caggcaatgc atcaggaggc acatgatgtc tcaacccctc40021attggtgatt gtaattttga tcatttggtt aaattactgt ttgcttaagt gttaactatt40081tttagatatg acgtatcaga gtgatcaata ctgtgaagcc tcttacaaca gtgtttagat40141tgaaaaatta tgagtgaaaa gcctttcaca gtcattcgga ttgaatatgc ctaacatgaa40201atccaaactg tttcaacatt tctttcaagt cattttctac tatggaaata ataatgcatg40261atttagccat gcctgagttt tatttcctca cgcaggcatt tatttcttta aaaaatttaa40321tttaaagttc actataaaac ttaccatttt aaccatttga agtgtacact tcagtggcat40381taagtacatt cacaatgttg tgcaaccatc actaccatcc atctccagga cttctttatc40441atcccaaaca taaaccatat ctactaaaca gtaactcccc aattccttct ccagcccctg40501gtaacctcat tttatatgcc atcacaatat tttagaataa acgccaactt cctggggtaa40561gatcatttga cttttgctcc cagatcagaa ctatgataaa aatattattt gctatatact40621aacagggaaa tttctggatt ttggcatgag ttactgtatt ataactctaa atcctggtaa40681gggaagcatt tttattctga aagtaaacaa aaataaccat tagtatgata atctttacct40741ctgggggtgg ggtattttgg agggggtgag tacaaagaga ggtgaaatta ttcctctagg40801gtggcagacc agagtttaaa ttgtttctta Kgtttttcat cttggttgtc taccaattgg40861atatagtcac tcagatcatc catgtttttg tacctggttt gataggtata aatacctata40921aattggaata aattctagca attagaggtc catgttcctt ggtctgtaaa gtgaatataa40981caaaactgac ttctctccaa agggtgcttt gagacaaaac tataaaacac tttggagagt41041agaagggggg tccttatttt tagtaacatt cctcagtgtt cttcaagaac tcacctactg41101ccaatcattc ctctgagtta agaatatata ttagaactag ttttaatagc actataaact41161atagatagca tggccatatg tccctgtttg cctggagctg aataaattac aggttcaaaa41221atactcaaat aataaaattg cctaggtggt gacccaccta gtttgttgct gtatggtgtt41281gggtatctct tttttttcct tgacaaattt gaaaaggcct tgaaaccatg agcagattcc41341ttggttcaga tggtctgtaa taggagacac caggacaagc ctgagtctcc tgctcttgaa41401agattagcat attgatagat tgcRtaccca ttatcagtta ggtatctgcc aaaccatttg41461cagcgtttct ctggagttca tacagttggg aagacttcct agaggcaaga ggtcttttac41521ctgaagattc agcagttact ataggggcct tcttttggct tcttgctctt gggacacctt41581tacttgaatg aggtagtctt ttgccattag tgcctgtggg gccttaatga ttcctattgg41641gatgagttgg ccagaagtga ggctctccac ttcatgttga aactaacttc tgcctgagta41701ttaaggatgg gtgattctga ggtcctgagc cagctggaag gactttcatt cactttctac41761tcttgtctga agaatgctta ggctacctta ctgttacttc tctggtggta ccaaaggcca41821tgtaatgaca tggctggatg cgatcactca cgtctgtaat cccagcactt tgggaggcca41881aggcaggagg atcacttSag cccaggagtt caaccagcct gggcgacata ggaagacccc41941catctctaaa aaatttttaa aaaatctagc tgggcatgtt ggtRtactcc tgtagtccca42001gctactcggc aggctgagat gggaggattg cttgagctta ggcgattgag gctgcggtga42061accatgatca caccactgca ctccagcttg ggcaacagag caagaccctg tctctaaaaa42121aaaaaaaaaa aaacaaacaa aagatgtgat gattgcctct ttcggttata tcctgctgcc42181taagagccca ccccagaatt tagtggctta ggataaccat tttcatagtt ttgtcagtga42241gggattaggg aagatgtcag ctgagctctt catttctgct ccagatggca ttggttgggc42301aatgggactg gaggttctaa ttctaagata gcttcttcac tcttatgaat agctcctcag42361tgtctgtggc cagtctctct ccatgtggta tctcatccac cctggccttt ccacatgcct42421taggcttccc acatcatggt ggttcctggg tagttactct tcttacagag tagggatgaa42481atgaaagcct ctattctaaa ttcagttaaa tactggtaca aatatttagt actgggattg42541tgggatcatt ttcttatatc ataattataa cccaatcagg gtagaaaagt gaaagaaaga42601aggacacttc attttctttg ctctttgtgt ggatgatggg ttgtacaaat agcaggaggt42661tcaacttttg gcttgttttg gattactgag tactatcctt cagaaatgat ttcctaatac42721tttcccggcc atctggcatg agcacagaag accttttact caataaaagt aagctaataa42781taataagttg aaacatcttg ataataaata tatttgtaca gcttttcaaa tcccaggtta42841ttgaaattct gtcagccttg gctggtttcc atgctccttc tgtgcccttg tgtttccttt42901ttttcttttt ttttttgttg agccactcct gctgacttcc aaaggatgct ctgagctcta42961aaaggaggag cagttttgct ggcatgcaga agtggtgaag gtaaggcttg gattcaatta43021cagtaatctc agaagttctt ggtagaagag gggcacagaa tcccagcctc cattttgcag43081gtaaggaaga agaattataa aaagataaaa tggcttttct taatgtcagc atctgattct43141ttaccttctt tcaccctagc aaagattgtt aacataaaac agatgttcct tggacaagag43201aaaagtaata gagctggatc ttttcatgat cctgcattta ttatttgtag cactggatct43261ctaaactagg aaaagcaatg tcgattctaa ctagaaatta tggggcattt aataacacgt43321gaggcaaaat tatgaggaaa ggcaagaaac caagtggctc ctagcttcta acaccttatt43381ttctatggtg taagttaatg aatgatccat ttttaatgcc ataaaaaagg aaatttccga43441taggctttac cttaacttca cgttgctgtt tatattacag acaaaatgct aaagttaaca43501agtgtaaaat gccctaggtc agggtttcga gtgcccataa gcaattctct agaattattt43561tgagtgctca gattatagat ttgcccccat accatatact gtattaccta tcactgcaag43621attctttata gggtgtcaag tatgtgggag gatttaaact aattgtctta aatgcttcat43681gggataggag atgatccaat gggaactgaa tgttttcaat taggatgttt ccatggatac43741tattaagcat gccaagattc atagccacag gagattttta cttacttcaa tattgattaa43801gaaatctgag aacaatctca agaagtgtag gatgtaggcg gaggataaat tttgaggata43861gtctaaaatt tgctttctct tccatagtta aggcctagtc cagaaaaagt ctaggcagtc43921agccaagtct aacagcagga tttatgtcaa gagaatgtcc agaataaaag tgaacatggg43981ttgtttcatt taccccaggt gtatttatta acatttattt aatgttactt attatttgtt44041tattaccatt agctttctta tgtactcggt acaaagcatt agaattacaa ccctttgtat44101ctttgttgtt tatgaggaaa acatcctaaa tagtgtctac agacatctta aaattcacat44161tcctctgaga aaggtttatc ccttaggtta ttactttttt ttttttttcc acacatgggt44221tttttacctg ggttctcaga tctccaagaa atcggtggat agaattcaag gtgacttctt44281ggatggagaa aaagtacgtc tttattttga ccaaccatgg atatttagca ttttcatcaa44341ttaatgaatg caggtgacaa atcacagtac tattggcccc atctttgagt ttgtcctcaa44401tagctcacag atattttcat attgtatcac agttgttgca gatatcttga aatattgttt44461attctaatca ctccttcaaa atttgggtgg ttattggcat tatcaccagt tttttttttt44521tttttgaggc agtctttctc tgttaccccc aggctggagt gcagtggtgt gatcttggct44581cactgtgacc tctgcctccc aggttcaagt gattctcccg cctcagcttc ctgagtagct44641gggattacag gcacatgcca ccacggcccg gctaatttat ttatttattt attctttatt44701ttttattttt aaattttttt tagatggagt ctcactctgt cgccaggctg gagtgcagtg44761gtgtgatctt ggctcactgc aacctccgcc tcccaggttc aagcgattct cctgcctcag44821cctcccgagt agttgggact acgggagcgt gccaccacgc ccagctaatt tttctatttt44881aagtagagat ggagttttac catgttggcc aggatggtcg cgatctcttg acctcgtgat44941ccacctgcct cggccaaaat gctgggatta caggcgtgag ccaccgtgcc tggccacatt45001accaccagtt cttgatactt aatgccataa caaaaggcac atgtatcact atatcacaaa45061ttagcatttt ataatatttt aataacgttt tcaatataag tggcttccct tgtaatcata45121tattttattt ttgcacttaa aaatataatt cctaaaaggg atccataaag cttcaccaga45181ctgccaaagc gtccatggca caggacgtta agaaccctgt tccaaatgga atgaagatta45241gaagcactat catatgttca ttttctgatc gcagtttaag aaagagtttc ctacttaagt45301gcttttcaca gaaaatctaa acttaaaggt taaaagggaa ttcttttgtg ctggctcatg45361tgtgggccct acaaagtact ttgcagtcat atctgacttt cgtttgagag tttgtcttca45421gaaatggaag aagaaaaggc attcaggtat atgacaggtt gcattctgta gaacattgtt45481aaatctatgg gaggggagaa aagttccatt cagtgtcttg tggattgtgg taggagtcag45541ttgtgttctt gcagaaattt ccctagacag tgcctcttgc ctcctccttt cttgtcccac45601agtactccat ggagcagatg tgggaacttg gatgcctgat gggaattgta accccatgct45661ttgccttatc ctggtgggga ctgatctgag gtttatccat taaactgtat cagttcatgg45721ctgggagtag gaatgcagaa aacacaagca aaatgactgc aggaaggttt aaataaaatc45781tcatatgtgg cttttcctta acacattatt ttgcaaaaag tggcagagtg aatttgctgt45841gatatgatag gatcattctg gacttttact taatttacgt gttctctagc agaatgctga45901aggagcacac ataaggagtg gggagttggt atttggttct gtgacaagtg ctatcttata45961acagaaaaat acatattttt gtgaaacttc aaatgcattt ttattcccca gcacatctgt46021gattctggat ccactacctt cttccaagtg tgatcaagca ctgtacctga cagtatcgat46081tatccttacc cgtaggcaaa aattgtatta catttatttt acagtgatct ggaaaaaaaa46141ggatgacaac aaaaaccaga cacacttcat ttctattccg gtaacatttt ttggtaataa46201tttattactg tacaagtcac acaccatata agtcacacat ttaaagtgtc aaattaaatg46261gtttttagta tattcacagg atcgtgccat catcacagtc aaatttagaa cacttattac46321ccccaacaag aaacctcata cgctttagca gtcaccctcc attgcccccc agttttctaa46381ccccaggccc taacaaccac taacctactt tctgtctcta tgaatttgcc tcttctggcc46441atttcatata aatggaatta cacattttgt aaccttttgt gactggcttt tttcacttag46501cataatgttt tttaaggtct acctatatta gagtatgtat cagtacttta ttccttttta46561tggccaaata atattccatt ttctggatat accagatttt gtttatccat tcatctgttg46621atggacattt gagttgtttc cactttgtgc tattattatg aataatgctg ctatgaacat46681tcatgtttac atcatgtata cattttaatg tggacatatg tttttagtta tcttgagtat46741atacctagga gtggaatcat atatatatgt ttttaaacca aaatcaaatt ttaatgttgc46801agaatccctg aagcactttt aggaactgtt cacttccata gaacagtttg caaatcagtg46861ttaaagtgga gagagtgttg atttttttgt gcatgtattt ggtagacatc gtttgaatct46921tgattttata ggctgtgtga attttgtcaa gttacctgtg aacatctttc ttctagtata46981gaatgttagt acctctggat gtttcacgag ttgatagact gtattattgc ttgaaaaatt47041cactgtccat ttattaccat caactttggc cacggatttg ctttggctag tggcctgtga47101ggggaagtga tgtgtgcctt cttccaggca gaagctggtg gtttgcctca tctcttttcc47161tttctgccat gatgaccatt gatgttccag ttggacacag gctccatcag cttgtgtatc47221agaatgaaaa tgatgtgggg cagagccctc atagattttg ttgcatgagt atggaaaaag47281tgtcatttta agccgtcaag atttggtgac tatttgatac tttagcacag tggtcctcaa47341gcaatggcag ttttgcattt ctcacactga gatatctggc aatgtctgga gacatttttt47401ggttatcaca catggggaag tgctcttggc atttagtggg cagatgctgc taagcatcca47461acactgcaca ggacagctcc cacaacaaag aattatctga tccaaaatgt caatagtgct47521gaagtttaga aaccctgctt tagtataacc tagtttatcc tggttaagat agggtgttga47581gaagattaaa tggaaaaaaa atatataaaa tacagcacac agggtggtct ttttttggta47641attatgtgac agagcttggc aatggctctg aggtcttccc ttgcatttgt gggtcttttc47701caacttccct agggttcagt attcagttta attgtagttg tgtttataaa tcaaggctcg47761tttttagcag aatcatatac cctttaaaaa agggggagaa ctaacattag atgcctgttt47821ggcaccaggc tctttgtatg tgttttgaat ttttacagat attctctgaa actgatattc47881cttacctcca ttttgttttg ttttgttttg ttttgttttg agacaggatc tcactgtgtt47941gcccagactg gagtgcagtg gtgcgatctt gtctcactgc aacctccacc tcctgaattc48001aagcaattct cctgcctcag cctcctgagt agctgggatt acaggcatgc gccaccatgc48061ccagttaatg ttttgtattt tcagtagaga tgggatttcg ctatgttggc caggctggtc48121tcttaacttt tggcctcctg tgatccccca gccttggcct cccaaagtgc tgggattaca48181ggcatgagcc accatgcccg gccctccatt ttgtttttga tgaagacatt gaggcactga48241agttaagtaa tttgaccaag atcacatggc ttataagtgg tgggcctaga attcgaagca48301catctgtttg cctccagaac cctcttcttt ccacactgct atactacctc tttagcagtg48361actttaacat agagaaatga aggtaatacc tgatttctaa caattatgtg ttgtattagt48421tatctattgc tgtataacag gttacctcaa aacttagcac tttaacacaa caaacagtta48481ttgtctgaca tagttttgag aatcagtaat ttaggggaag ctaagctggg tggttctggc48541tcaggatctc tatggagatt gcaaacaagt tgtcttccag ggctgcagcg atctgaagac48601ctgattggag caggaagatc cacttccatt gtggtgcact cacagggctg ttggtgggag48661gcctcagttc ctcaccatgt gggtctctct ttagggttgc ttgattgtca tcatgacatg48721gcagctggcc tccttcagag tgagggatct cagagagaca gcaaggcaga agccacaata48781tcttttgtga cctagcctta gaagtcacag atgatcactt ctgctctttt ctattggtca48841catacacaac cctgatataa tgtgggagag gatgatacaa gggcatcaag aatactatga48901gatggaaatc actgggggcc atcatggagg ctggttacta catgttaatg atcctcattt48961gccatagtta caaaattatg aattatattt ccttttcata ttctaatatt ccaaacacct49021gttttgtact ttaagatact taaatcattt agaatgtgag tgaaaataca aattggcttt49081tgtaaacact tactgaatgg gtaaaataaa ggccgtggta cagatacttc attggggaat49141ttttttttta aataggtttg actagctgag gcttcttgaa tcctgaaaag tttggtctga49201tgtacttttt tttttttcct ggaaaatctc cttgtaccaa acactgaatt cttcatttgc49261caaagtccac ccaactttct gatgttttta ttgctggttt gatgtgcttg aatgccagcc49321agagtataag gtgaatggta gatgcaagga ctgtgcagtt tccacggaga cttctttcag49381gagcccttca ttttttcatt cagctattgc tgggaggcat ccccttccag ttaatgtgtg49441gtacacatca atttgagtgg ctcttagaaa ggggctcagt ttatacaaat taagaattga49501ggtttttttt ttctctgtat atcttcaacg ttaaattaga tttctatgaa acttgaagat49561gttaacagga aatcttcctg aaacagttta actatacttt tgtttattac acttgatttt49621gagcttttgg gaataaaagt agggtcctgg ctaggtacag aggtatatgg atttactgtg49681tctaatcgtg caaccatgaa ataaacgttt gcacttcagg aatcaaccca tcacttttta49741accactgttg tttgcattgg cttagagctt tgtgcttcct tcaactattt tccttttttc49801tttcaaaact gtgttcagtt ggttgtgacc aattttacac agagactggc tcactcactt49861atgttcttga ctcctgctgt cccaatatca tgcttcattt acacagggat agcttttcct49921ggctttatct gttgacttga catcctgtgc atcatatttc ttttcttaga tatagagaag49981actgaaaaaa atccacgatt tgaagaaaat ttRcaaaagt gcagagcaat gtgaaagaga50041tagctttaat tttaattttc actgtagtca gaataggtgg cctcctgatg ggcagacttg50101ctttctcatt ttactctgtc cagtgtagat acccagatca atcaaggtca agctggtctg50161gtcattaata caacttttgg ttttaaacac agctgtgtac gggggttctt agcattatta50221atagaggtta taatgaaaac aaggtatagt catacagttg tctgtgtggg tagtggcatg50281gaggtggagt tgttgcttaa gatctgaaag tcaggaagct ggaggttttt gtaattttat50341tcattatcaa tcttaagaat ttattttgat cattataaat gcatgactgg tgctattttc50401caatcctgag gttttttggg aaattttttt gaaaagaggc caggatgagg ataaaaatta50461attttgaaaa tccatttcaa agactcatta tgctatgctc catcactgca attttaatgc50521aacttatata ttgtgctttt acctttggga ctagcattct taaaagctga cttttaattt50581ggactttcta atatgtgtcg gctctgtcag ctcacgagtg tgtttttatc tStgcatcat50641gttatattgg gacctgggtg gaatgtaaat cttttactgg cctaaatctg agtctaatcc50701tctgcatggc tttggtagaa aatcaatttc ctctgctctg tcaagatttt tcaggtcagt50761gctctttgga agtttcccag ttctcttaga ttaaaataga ttttgtttct gtgctacata50821ctggtattag ttagtaggac actgggatct aaaagcaaga gaacagaggc agacaagata50881tccatgagtt cttctatttc agttaacttg ttgacacact gcacctttga ggaaccaagg50941agtttgtgtg ctttcaggca tcatttgggg gagcaattgt aaatgactcc tgtaggtgag51001gctgtgcact taattttaat gagttcaggt tcattaagtg acctgctcac tgggtcatta51061gaagtgacct cactagaagt gaatggtaaa gagaagaact acctaagttc cactcagctt51121gtttcactac tgtggggtat tggaagtaga gtgacaactt ggactttctc agttttagca51181ctgaaaagtc ccacatccca ggaacttact cagtccccag caaactaggg ggagcggggg51241atggttggtc cccctagcag tgaaaagtaa ggggatggat gatttggaac tggttagtac51301ccttatagat tttttttttt ttttctgaga caggttcttg ctgtgtcacc caggctagaa51361tatagtggca tgatcatggc tcactgcagc cacgatctcc tgggtccaaa tgatactccc51421acttcagcct ccccagtagc tggaactacg gggacaagcc accacaccca gctaattttt51481atttatttat attaattaat taatttattt ttttgtaggg acagggtctc gctttgttgc51541ccaggctggt cttgaactcc tgggctcaag tgatcctcct gccttagcct ctcaaagcac51601tgggattaca ggtgtaaacc accatgcctg gctgtcttat aaatacttta aggagggatt51661tattagagac atgtagcacc ttgctgccca gtgatgctag taaatgagtg actctacttt51721gatttgacag aagaaacaat ggaaagagga gagatggaag tttttctacc taaacttcct51781aagaggatgt gtttagggga tgtgttggta aaaagttatt agcattaatg tttacctgta51841tctttctcat agataatgca aagaccttta actccactta tccctccttt taacttatgt51901tgttattgcc ttgtatttta atactatata ttaaactcta caagacatca ttattttatg51961caggcattgt ttatttaatc agagttcagt aaactttttc tgtaaatgcc agatggtaaa52021tattttaacc cctgcaggcc ttatagtctc tgtcacaact actcaactcc actcttaata52081gtgccaaaat ggctatagac aatatggaaa tgattgtgtg tacccatgtt ccaataaaat52141ttatttacag aaacagacag tgggccagat ttggcctgca ggccttactt tgacaactcc52201taatttagat ttatctacat atttatttat tttcccttta tttcttgcta tatctttgag52261gttctacctg ggatcatttt caaggcaagg atgtctattc tcatcactgt ttttcagcgt52321tgtgctagaa attccaacca gtgcaataag gcaagaaaag gaaataaaag gcatacagag52381cagaaaggaa aaaaaaaatg aaactggcca ggtacagctg ctcatacttt gagaggcgag52441ctaggaggat tgcttgagcc caggagttcc agaccagctt gggcaatgta gggcgacccc52501catctctaca aaagtaaaaa taaaaaaaaa attagcaggg tgttatggtg catgtctgtg52561gtctcagcta cttgggaggc tgaggtggga ggatcacttg ggccttggag gtgaagtgag52621ccatggtgac tgtatcactg cattctagcc tgggcaacag agtgacaccg cctgtctcag52681aaaacaaaac aaaaacctgt ctctatttgc agatgacatg attgtctaca tagaagaccc52741tgagaaatct acagtctaca gaataactcc tagacctaat taagtgagtt cagcaaggtt52801tcaggatata agataaacat acaaaaatga gttgtatttc tacatactag ccataaacaa52861atggacactg aaattaaaaa tacaatacca tttacattca ctcaaaaaat acttgggtat52921aaatctaaca aagcatgcac aggacttgta tgctgaaaac tatacaatgc tgatgaaaga52981aatcaaagtc taaatagatg gggaaacata ccatgttccg ggtttcagga tataagataa53041acttacaaaa atgagttgta tttcacatac tagctagaaa caaatggaca ctgaaattaa53101aaatacaata ccatttacat tcactcaaga aatacttggg tataaatcta acaaagcatg53161cacaggactt gtatgctgaa aactacaaaa tgctgatgaa agaaatcaaa gtctaaatag53221atggggaaca taccatgttc caggattgga agactcaaca tagtaaagat atcatttctc53281ccccaaattg atatacaggt ttaatgtaat tcctgtcaaa atcttagtaa gatttttttg53341tagatataat tataaaattt atatggaaag ggaaaagaac tagactagct gaaacaattt53401tgaaaaagaa taataaagca ggaggaatca gtctgtaaga ttttaagaca tataactaca53461gtaatcaaga ctgtgtgata ttggtggagg gatacatgta tagatcaacg gaacagaaca53521aagaacctag aaatagacca acagaaatgt gcccaattga tctttgacaa aattgcaaca53581gcaattgaac ggagaaggaa agatagcctt tttgacaaat ggtgctaggt caattaaaca53641tcaatatgtc aaaaaaaaaa aaaaactctt gctgtaaata gtacacgtta tatgaaaaat53701aactccaaat gtatcacaaa cttcaatgta aaacataaaa ctataaaaat aattagagaa53761aatcttcagg atctaaggct aggcagagtt cttagacctg acaccaaagg tatgatccat53821aaaagaattg ataaattgga tttcatcaaa ataaaaaata cttactctga gaaagacctc53881ataagaggat gaaaagacag actacagagt ggggggaaat atttgcaaac tacgtatccg53941acaaaggact ggcatctaga atatataaat aactttcaaa acttaacagt agaaaacaac54001ccagttagaa agtggaaaaa atacaagaag ggacatttca ctgaagagca tatacagatg54061acagataagc atgtgaaaag atgttcaaca taattagcca ctagggaaat gtaaattcag54121accaaagtga gctatcacta catacctata agaatggcta aaattaaaaa tagtgacagc54181atcaaatgat ggcaaggctg tagagaaatg gaatcattca tactttgctg atgggatgta54241aaatgataca gcccctctga aaaacagttt ggcagtttct taaagacact aaacatgcaa54301ctaccatatg acccagcaat tgcactcctg ggcatttatc ccagagaagt gaagatttat54361gttcatataa atgccagtat gtgaatgttt tataggagtt ttattcataa tagcccataa54421ctggaaacaa tacagatgtt ctttagtgga tgaatggtta gacaaactgg tatatccata54481gcatggaata Ytacccagca gtaaaaagga gtgaagtact gatacatgca atgatctgga54541tggatctcca gagaattatg ccgagtgaga aaagccaatc ctgagtggtt taaataccat54601atgattccat ttatgtaaca ttcttgaagt gacaaaatta tggagatgga gaacagatta54661gtccatgcca ggggttgagg gggcaagggt tgggagtaaa atggatagct ataaaaggca54721tgatgaggcc aggcgtggtg gctcacgcct gtaattccag cactttaaga ggccgaggcg54781ggtgaattgc ttgaggtcag gagtttgaga tcagcctgac caacatggtg aaaccctgtt54841tttactaaaa atacaaaaag tagctgggtg tagtggcagg tgcctgtaat cccagctact54901ccagaggctg aggcaggaga atcgcttgaa cctgggaggc agaggttgta gtgagctgag54961attgtgccac tgaactccag cctgggcaac agagcaagac tctgtcttag gaaaaaaaaa55021aaaaggcatc atgggtcctt gtgagcatga aaatgttctc tattgactgt gtcaatagag55081acacacaatt atgttgtgtg tcacaaccat aatcacaaca ttatggttgt gatattgtac55141tgtagtttta caagatgtta ccattgggag gaagtgggca aagggtacat aggatctctg55201ggtatcattt cttacctctg catataaaac tacagttgtc tcaaaaggtt taattaagaa55261agtatttaag aaaaagaacc ccaggatcta aaaagatcag agtcagagaa tcacaaatgc55321taaataagaa gcacttttgt atttggtgat gttctgaaat tctggctcag ttttgttatg55381aaatggagcc acgtgggtcc atgtgagtga gtccctgtgc aYaggtgtat gattgacttt55441ttccctcaag tgcttctaat ttgggatgtt cttcactgtg tgccctttga gtgtctcttg55501cctctggttt tttgttagac aaaaaaacca gtttttaaat gtcagttcca aggtcttttc55561tgggtacggt taacttttat gatttcattt atgcagtggg ttttcaccta agggccatgg55621tttctagaca ctagactcgt ttttcccaag cgattcatct ctttgcctgt gggatggaga55681gattcattct cttcttctct tttgcagtag aaactccagt tttgagctgg gcacctggat55741gcccagagta aaggttgtac tgcccacatt tcattgtagc taggtgtgcc tgcgtgacca55801agtatgaacc ttctatttag attaagccac aagataagtg gagctatcaa ttggctttaa55861caaatattta gtgaataggt aaaataaaat actttatata agagctgtgt gttctgggtt55921gaattgtgtc ccccacaaaa agatatatta aagtcgtaac ccttagtacc tcagaatgtg55981accttatttg gaaagagggt ctttgcagat ataattaatt tcaggtgagg ttataccgga56041gtagggtggg ctcctaatcc aatatgatga gtgtcctcag aagaaggcag tcaggcacat56101gtaccctaaa acttaaagta gaataaaaaa taaataaata aataaataaa taaaaaagaa56161ggcagtcagg tgaagacaca gacacacagt gggaaccccc tatgaagatg aagatagaag56221caatgtatct acaagccagg gaccgcctgc ggctgcaaga agccaggtga gacacatgga56281gctgattctc cctcacagcc cttagaagga acgtgccctg ctgacacctt ggttttggac56341ttccagtctc cagaaccatg ggataataaa tacatttctg ttgtttgaag ccacccaatt56401tgtggtactt tattatagca gccctaggaa accaatatac tatgcttttg atattatttt56461taggatgttg tggtttgaat atttgtgtct cccccaaatt cctatgttga aacctagtct56521ctgatgcgat agtattaaga ggtggggcct ttaggaggtg catgagggca gagcccttat56581gatgggatta gtgttcctag aaaagagact tgagggagct tgtttgcccc ttccagtatg56641tgaggataca gcaagaaagt gccatctatg aggcagagag tgagccctca ctatacactg56701aatctgctgg caccttgatc ttagacttcc cagcctccag aatggaaaga aatttctgtt56761gtttataaat tactgagtct aaggtatttt gttgtagcag ccccaacaaa ctaagacatg56821gagtatttta aaaaatgggt tgaacattca gaggcttctt aatctgagtc cttagatatt56881tggactgact ctgccttctt tttccagtga aaagatctcg ctgtatgaaa catagacatt56941cttcattgcc ccaaagccct tccaccttcc ttatgttttt attgctggtt tgatgtgctt57001gaatgctagc caaagtataa ggtgaatggt agatgcaagg actgggcggt ttccatggag57061acttcttgca ggagcctttc ctttctcagt tcaacagttg tcaaggaaac atctagtttc57121aattaaataa atgtttgtgg ggtctgccaa tttgagtgtc tcttggaagg ggatttggtt57181aaacttctta aacatcgtaa taaacaaatt cagagcagag gctttttagg ttcccttcca57241gttatattag gtttttctgt aacttaggga tgttaactag tcctctgtct gaagtgatca57301gactatttat tcatatccag aaaatattat ttttgagtct tgtggggagg aaaactgagg57361cttaaagaaa gagaggccaa agccagggag ctggtgttac atggcaccag ctatatatac57421atgactatag tccatgctct ttagtacgtc ctgcaggata agagatccaa ggggcctgac57481ctttacaaac tgtttacagg ctatggagtt tgctggctca gcactgtgtg tttattgctt57541ctagttagct attgttgtat cagtggctgg agttggtgat ggttcacgga aatagtggtg57601agagtacaga ttctaggtat atgcaattct aataaatgag acttccctag tgttatctgt57661attgtgacag cctccccatt gtgctcttta tcaaacagta aggtatggta tattgcaaaa57721tgctcacaat actttgcaac tcttcccatc aagaggtaga gtcagtttct ctattcttag57781gatcttggct gtcctgttga tttggtctga ccctggaata cagtagaaaa gaccctgtgt57841gggttttgat cctaggcgtg aggtagagct tcagtttctt gctgctttgg gaactctgcc57901accttgtaaa aaaagccctg gctggcctcc tggaggaaga gagactatgt ggaggaaaat57961gaaggtaccc agacaaccca atagtcaact gcagaagcat aagtgaccct catctgagat58021caatggtgcc tggctcggat cagtataatt gtccagctga actcagccca aattactgac58081ccccagagtt atgaattaaa taaatgctgt tgctttaagg cacttaagtt ttggggtggt58141gtgttataca gtagtagata attgatacag ccaagtttgg aagctctgga atgatcaggg58201ctcacctttg acctgtcctc tataaaaccc aggattctta gtaacacagt cctgttaata58261acagaacttg cctatggcaa atctaagcgg aaaagcagtt tattggcagg atattgtgta58321gctacatctt tctttaggaa gaccagggac aaggcttgaa aactgcacaa ctagggggca58381gtactgccag aactacagcc agataacctc acagaattgg tagctactga atattggatg58441cctccaccta caccaccaac attgcctgca ttggacattg ggtatggggc tgctcctgga58501actggatgtt gttcctgtca ctagaaaaca atcatctact gttcttgctg tttcacagca58561ctcattactg attcagatgc ccgtgtgcca cgtgtttata gcagtgcaat ttgtaattgc58621aaaaataggg aaccagccca aatgcccatc aatcgagtgg ataaagaaaa tatatcaaaa58681atatatttta tatgtgtgtg tgtatgtata tgtgtgtata tatgtgtgta tgtatatgtg58741tgtatatata cacacacgta catatatata catatatata cacacacgta catatatata58801cacacacgta catatatatg cacacacaca tatatataca cacacgtaac atttataata58861cgccatggaa tactactcag ccataaaaag gaatgaaata atagcatttg caacaacgtg58921gatggagttg gagaccatta ttctaagtga agtgactcag gaatggaaaa ccgaacgttg58981tattttctca cttataagtg ggagctaagc tatgaggatg caaaggcata agaatgaaac59041agtggacttt ggggcctcag agggaagggt gggaggtgag ggataaaaga ctacacattg59101ggtacagtgt acactgctcg gctgatgagt acctcaaaat ctcagaaatc agcactgaag59161aacttaactc atgtaaccaa ataccatctg ttcccccaaa gctatttaaa taataataaa59221aacaaaaaca aatgcctgtg tgggtggtgt ctggcagagc gtaggtcttg tgcccatacc59281ctgattgaaa agaaatcagg aaagtgagaa tctgacactt ttagcttatt tagctgaaag59341caggtggaaa atttcataaa cactggcaaa gggttcggat aacaggcagc caaaagaaaa59401atggcaaatg ttcattagtt tctgttgtat actctggtta gctcagttgg ttacagcaca59461gttatgatga agccagcatt gtggtttgtt cttaagtttt attgaaagaa aatagttctg59521agattgtccc ccttacacct gctccctctc tcacagatgt gtgctgatag ccatttgtaa59581ggccaggcaa tagtggggcc taacttgcat ctgtctctgg ctgctaaggt acaaagatta59641tgtgtctttc agttggtggg cccagagact ccattcactg gcatagtgtg ccacataaga59701gaactgaatg aatggtatct gtgtgaatct aaggggttgg ctaattgcgc agtgaatttg59761gttcatgtaa catttataat acaaattgtt gggcagcaag atctttcaat ccagacggtt59821tctagttttt cagaatttct ctgataactt ctctgaactc tgaaattctt atttgcaaaa59881ccatctatat agggttcatt taaatattag aaatagtact cataaagaga aaagcataat59941gcctagagga tgtgtaatag atgttcaata aatgttgatc atttccctca aacttcttag60001attcacacac cccaaccaga ttagaaactc tgacacggct tgcccttttt gtgttcaatt60061ctctctcagt tgttaagtga agacccaatc cagttgttaa aataataaat gcacctcttg60121cctccctgac attctggcaa ccttactctg aggaggctgg cccggttcga agctggctgc60181tccagggcac taaagctgga gaagggccct tccaccctgt tcataactgt ggacagtaaa60241ctgtatttgg gatagaggat cttgggaaca ataaggacct agatccaagc tcatgcattg60301tggtgggact gtcaatttag ggggatgggt agattttatc agatctgtca gtggaaagtt60361tggggagatg gaatagtatc tacaggtagt tccaggtagg aggaagagat ctacctggct60421cctagggatg tccatcctgg caagcattgc tcagtggcag gagttgtaag agtagggaag60481gtttaggaat ttggatatca gggaagtgtc agcaggtagc tggggatgag gtgcaggaca60541aagacagacg agaggaagac caagccttaa agtgatgagt atctagatag ggaagaggat60601ggatgggtat agcttggaca cagaaagggg attcagtcat ggaaagaggt cagtctgcaa60661tgtaatgtgg ctatcaggtc acaggagggc caagagcagg acaggtttgg ggcttggttc60721tggttagtcg aggcagagcc ccttagaggt gtttggattt gccatggtga tgcaggggac60781ctgagtgtgc tggttgtcat gagatcttgt tgctgggaga acagtcaaga acagggtaga60841aaggcccttc cccagcttta gtgtgccttg gagcaggcag ctttgaatag ggccagcttc60901ttcagagaga gctgccagaa tgtcagagag gcgagaggtg catttgttat tttaacaatt60961ggattgggtc ttcactggga gctctacaaa gaaaccaaat tagtgaagtt agattgccta61021tcccaccaac aatggatgtt tatggtatga gtttagctct gtgctagaga atctcaatat61081atttagattt gggcttgaga gagagaactt cagtgtgatc tgagactgaa cttccattca61141cattcattcc tcgaggaagc atgttgagga tctatgtgta atatactggt gttccattta61201cttgaaatat cctggcttgc tctattcctc tcacttaggc actcactcct ctaaaaaata61261aatgtgtata tgtgtatgtg catgtgtgtg tacatgtagg tgtaagtgtg cataaaatat61321ggaatagact gaaatcaaca aacttataaa aataccaaat aacctcaatc tgtaatttta61381tttttgagaa gcactaaaaa tgtcaataaa tgaattgtct gaaaaactaa tcctttggct61441ggtgagataa gtctgagttg ccagaaccag ttaaagcttt atataatttg gcagtacatt61501tgcgttcacc tgagatttct ctgcctccag ctgtacaaaa attctgtgat atgcagagcc61561tcgaaacctg ggccagaatg ctaaaactaa ttttctttcg taatctgagc cgtaattgca61621ttttaactca gttctagtgt gaagactgct tttgtatgtg tgtgtgttta cttcatgtca61681ttaatttctg gcatttaagt tagtgcctgg cacatagtct accctcagta aatgttagtt61741cttactgccg agtctactgt tctttccttc taaaccagtg gggtagatgc ttcctttgtg61801cctcagtggt tcattccctt gccctcttcc cctctcctca cccttgagtt gttagatcct61861ggttttgccc caggcaggtg ccaaccccga ggaactcctg accctgtggt gatgcatgcc61921tgcatgtgtg cagccagctg ggcagcttgg aaagccatag tattgacttg ccaaataagg61981actagtttac aaaagtcata agagatcact gtcatcaact gctatcacta tctattatcc62041tttctctttc caaaacacaa tttaggtgca taataaaaac atctgtagaa ttactctgtc62101aagcctatgt atgcccatca gttgtgcaag agcctcagtt cctacttcta atgaatttta62161tactgattat cccagacctt tctgccagct ccctaatagc agaggagaga atacactgtt62221ggagtcaggg gcaaccaaag cagctagtct gagtatagaa tttctttgaa ttctcatctt62281taagaaccat gctcagctgg gcacagtggt tcatgactgt aatcctagca cttcgggagg62341ccaaggtggg aggatagtaa tggcctggtg tggtggctca tttccataat cccagcactt62401tgggaggcca aggtaRgcag attgctggag tccaggagtt caagaccagc ctgggcaaca62461cagggagacc ccccatctct acaaaaaaac aaaaaagtag ccaggtgtga tggcacttgc62521ctgtagtccc agctatttgc agagggctga ggcgggagga tcacttgagc ctgggaggtt62581gaggctgcaa tgagctgtga ttgtgccact gctctgtagc ctcgaggaca gcgtgaccct62641gtctaaaaaa aaatggatga tatatccaca tgattcagaa ttgaagaggt acaaaacaat62701aaccaataaa ctctttctcc ccactgtgtc ctagccacct tgttttcttc cctggaggca62761tccagaagcc aattttgtta gtttcaactt ctggagatgg cttatgaaaa tataagtaaa62821tgcatctata ttcaacccca ttttagaaga aatgatacct actatatcag ctgctttctg62881tgcaccttgc tttttttctt tttcacactt tagcatgatg atttttctgt attggcacat62941aaagagcttc ttctttgtta gttggtggtt cttatggcag acaactgaat ccaccctaga63001tggttaagcc aaaaagggat tcggatttta ggtaactcat ggaatccttt ggagggctgg63061aggaacaggc tagcatccgg gtgcccagaa acaaagcccc aagacacacg tagtactagc63121attaagtgga agccacctgt gctgctgcct ccactgaaca gcagatgcta ggtatttgac63181ttgactgcaa ctgcctctac ctccactcct gcaccgggaa ggcaacctag cagtgacacc63241tgcaaaagta gaagctctga gcagagcctc tttccttaca tttctcactt ctaaagcaaa63301atctggtgtg gatgtatctg attagtggtg actaggtgac acatttgcat cttagcttca63361atagagtcta ggaattttat tctcaacttt taaacttgga ggtgggactt gtaataggtg63421aaatttttgc aaatagagaa ataattttca aagtatgatg gacagtcaca gatatggcaa63481atgtccacta gagctgctta gaattccatt gtatggatca accacaattg attaaactaa63541tatcttagat ggtagacctc taagtgtttt ctagttttct agtctgtgac ttaggcaata63601ctgtattaat gaccatatac atgtgtcatt ttgctaatgt aaaatggaat ataaatttct63661agcagtggaa ttgctagatc aaaaggtaaa ttgtaattct tgtggatatt gccaaatggt63721ttctggatgc tatactgatt ttttgcttcc actagtgatg ttcctgattt tttttttttt63781tttttttttt tgcacagagt cttttccttt tttagattcc ttctgagcca gagtggcagg63841ctgagtgttc cctgctactg gggtgagttt tgggccttgg gccaagaatg ctgacaaatc63901agatctgggc cataacctat gccaatatag actgtattta gcatctggac tggatttggc63961ttatagttta ttgtagtcct tctagcccta aattaaatgg catcttgggg tatctaaaag64021ataatgttgc ttccaaaatt gatttgcaat gtgggagtag cttacatctt ggattgttat64081gtgttcacaa atgcataaag atgttttcaa agaatggttc taaagaatta taataccttt64141tctttttaaa aaagcaaaat cttggccggg cRcggtggct cacgcctgta atcccagcac64201tttgggaggc cgaggcgggc ggatcacgag gtcaggagat cgagaccatc ccggctaaaa64261cggtgaaacc ccgtctctac taaaaataca aaaaaattag ccgggcgtag tggcgggcgc64321ctgtagtccc agctacttgg gaggctgagg caggagaatg gcgtgaaccc gggaggcgga64381gcttgcagtg agccgagatc ccgccactgc actccagcct gggcgacaga gcgagactcc64441gtctcaaaaa aaaaaaaaaa aaaaaaagca aaatcttatt tctagtgtgt gaactattgc64501atagattttg ccaatatctg aactatggca tccaattgct ctggttggtt gccatgaaag64561tttatagtaa cataggaagc tcctgtttga aagctcctga aaaatttaaa caaatgtata64621taaatgggtg aaaaatgggg gctgaggaag aaggtgaata catgtgtttt cttctcatgg64681aagtgttcac tattttttat taaatttttt ttagagacag ggtctcacta tgtttctcag64741gctggtctcg gactcctggg cccaagtgat cctcttgtct cagccttcca agtagctggg64801actacaggca cgggcctcca catctggctg tgtttgcagt ctttaagttg gcatttacct64861agtgcagttg attatttcaa gaaaatacac tttctacatg gctactgagg gtatctgaaa64921ggatataaag tatagcccag ggaagtagca tgttgttagg caggggttag taactctttc64981tggaaagggt cagttggtat tttaggcttt gcaggccaag aaacagaatc aaagatatta65041cataggtaat tctataacaa gagaaaaaat aaacttctac aagcttttta ttgacaaaat65101tcaaaacata ataactgagg actttttttt ttggtaatac aggtctactc ataagaagaa65161cagcattctt actttgggga taacattttg cttaatggga ttcaaagaat agtgttccct65221ttcatcaaat cgattgcaaa tgttcatctg taaataccat ttattttctc cttgtgggcc65281atagaaaacc aagcagtggg ctggatttga cccatgagcg gtagttttcc aaccactgat65341gtaatggaaa gaccatgagc tttggacctg gatgcatgta ggtttgagta ctggctctga65401atttgtgacc ttgaacatat cacttaactt tctgaattgg gaaggtttat ttgtgtataa65461actgagtgta atgacaccta tctcatggaa aaaatattac ccccatcttt gtgattttgc65521attagtattc tagttctgta gatgctgata tgatttagct agacttacct agaaaattag65581tatttaactg attttaaagc tgacttttca agggatagaa cactcacatg gtcttaaagt65641acaatgtaaa aacatagctt tgcaatgaag agatctggca gtcaccacat taatcaaatg65701accaagcttt gcaatcagta ccaatggtgg gatgttctga cattgtgtgc ccctcactgc65761tgtgtacctc tcactatatc tcagtatcac ctgtgtggtg ttcttgccag aaacttttga65821ctttaatgta atcacaataa aacagacaaa tccaaaacat gggatgttct gaggacaact65881ggcctggatt cttaaatgat tcagtgtcat taagaaccaa gggggattgt tctagattag65941ggggagacta agggaaaatg agataaaaat gcagtgcatg aacattgact aaatcctgga66001tgaggaaaag aaaattcagc tctgtaggca ttttctggat aatcaaggga atgtgaatgt66061ggaccatatg ttgtatgtta gtccattttc acgctgctga taaaggcata tccgagactg66121ggaagaaaaa gaggtttaat tggacttaca gttccacatg cctggggaag cctcagaatc66181atggcaggag gcaaaaggca cttcttacat ggtggtggca ggagaaaaat gaggaagaag66241caaaggcaga aacgcctgat aaacccacca gatctcatga gacttgttca ccatcacaag66301aatagcatgg gaaagaccga tccccgtgat tcaattacct ccccctaatt gttcaagtcc66361ctcccacaca cgtgggaatt ctgggagata caattcaagt tgagatttgg gtgaggacac66421agccaaacca tatcatgttg tatgataata ttgtgttaaa gttgatttcc ttgggtgtgg66481tgatgatggt gcggtggtgt acgaaaatat ccctgttctt agaaggtgaa tagtaaagta66541cttaggggtg gtgtgtcatg aggtgtgtct gtaacccact tttcagatgg tttggcaaaa66601tgggagggaa ggagggaggg agagagagaa tgtgtgtgag tttgtgtgta tgtagaatag66661gtgattctgt gcattgctgc tcagttctta gcatagcact cagcacattg cttttgatag66721ggaggtgccc aaccaaaact tgttaaacta acaagtactt taaggtttta gtaggattct66781ctgaaatcac acctcagcac tgcctgtctg ccctctcttt agcagtgtca tttggcttag66841gcatatacta taatgcctaa tgttgggtaa tcagatatgg gggttcatgt tttcttgagt66901agctcaccca tgtccctctg gactgaaagt ctctcttgaa tgttacagta taaatgaatg66961ggaaaggaaa acccagcata gaggttgacc tgcgtgttca gctgttccct gagaggctga67021ggtacaattt gtaagcatgt acttcagatt ctcttctgta tttctgttgc tgagtttgcc67081cacttgggga gggttgacac ctcctggggc tgtgctgagt aggaggtgca tctctggagt67141tctgactggc tggagcatca ctgaggtatt ctcctgggct atgagggcta gggccaggat67201aaagaatggt ggatgccttt ccttatgcct tgatcctaac cggtaccctc ttagctgagc67261aggaagggaa cagagaaatt aatattctct tatattcctg caggaactct tatggctata67321tggtgtgaga tgactaaagc caatttagca gctacttgta aaagtaataa aaatggaggg67381gaagaagaaa gattctgatg caacaggctg tgaagactaa ttcaacattt atttattgag67441ctactgagca tatagcaggc atagtaaagt atgagtcctg ggggatacaa ataataagaa67501tataccctcg ctttctgctg ggcacttagg gcatagggaa aggagatggg ctacagtgga67561gataattatc aaaataccaa ttgtgctgtg ttgcaaggct agggatgtgt tagaaagcta67621gtaaaaagtg ttatgggaat gcagagaaca tgccatgtgg aattgactac agtttcaaag67681ataaggttgg tcttgaggtg acttgaagaa tgagcctggc aggcaaagaa atgggcatct67741ggtgtgacta aagtcttaat tagctgtagg gtggggagga gagagacatg acgatgttgg67801aaaggtagtt tgggccagac aggagggcct cagaagccat gcttggtttc acaggaaggg67861cgtgaagagt cccagaagcc ctttgaagtt ggagggtgat gtggttttga taagaggtca67921cggagggaat gtagaagctg gtgtgtgaag gtgtgtgagt ctggagacag gacttgaaac67981agtcgaggcg agagcaaaga atctggattt tgctagctgc agcagaggga gagtagaaaa68041tggggtgctc ttggagaatt cttccagccc ctcctactag gagtttttac attttcaaat68101ctttctaagg aaacctttaa taatacttga gagaagctta tttaaacctt tgaacagttc68161tgaaaatgta ttacatttca gccaggaaag aaagtattat ttcaggcaaa ctggaacatg68221gttactcagc acagtcgcct aatcttatag aattaggtag gagggaagct atgttagtgg68281ccttcaacct tcttccagaa ctttgttcag atttctccca aatggtcggt gaatttttct68341gatttgtatc cagcatccca gccatttagt gtacatccgt aagctgtaag gaagttgatt68401taatctattt ctttttttca tctcttgcct tccacctgag aaaacactac tccataggtt68461cttattctgt attaggaatt tgcacaatgt gtaacctaag atggatccag ctgggagttg68521gatccagctg ggagtgttac aggtttggag tttagactaa agaagaggag tggaatcttt68581tcattttttt cccctttatt ttgaggaaag cccagtgatt tggattgttt gaaggatcaa68641ttatatcaaa tatctccact aattgagtat ctgggataaa cttttttttt caccttattt68701tctttagata ccattaagca tttcgaccca caacttacag cttttccatg gagattttaa68761acttagaaaa ctttgaacct tgatgaatga aaaaaacctt tcagaactaa cctttttttt68821tttttttttg attcagaagt ccatgtgtaa gttccaaagg cttataattc ttttctcttc68881cttgaggctt gactatttca gaacgttctt ggtatgtctc ttggaggttt ggtgtacaaa68941gagcaaggaa agtaacctat gggcttaact cacttatgtg tatatattct actttggctc69001caaagcaatg attctagtgg tgatatggga ccaagggaca gatttcaagg taaataaggc69061tgtttactaa tgtcagatta aggtattagt cttcaagtac cataaatgtt ttactagcaa69121tttcagttac ctgtagtaaa gagctttcca gttccatgtg gtagttatgg taaaggagca69181actgaaagtt aacttgatag tagattaaca ctcactgaaa atatagttgt tattttttca69241cataccaagg attacaatgt cttttttttt Ntctattttt aaaatagaga cagggtcttg69301ctatgttgcc catagtcttg aactcttggc ctcaagcgat cttccttgag gcctcccaaa69361gtgctgaggt tataggcatg agccattgtg cccggcctac aatttcttaa tattaggagt69421tattttgcaa gacaactatc tcaatgttgg aactacacta atgttggtgt gtttttttcc69481ttgtgtggga tagtctgtat cttaagatgt gtaatgtata tattcacaat aagggctaca69541ttataaaatt cagagacatt ttatttttga ggtaggtttc attcacttaa ttgagaattt69601gttgactgta tcttgggtgc ctatccttat getaggagat gaatggaagt tgccccctgc69661cattgaagag aagacatttt aatgggaggc agagataaac agataattaa ttataatgtg69721atacatttgg aaatagtcac aaaaaacttg aatatggaga ttttcaaaag tacaaagaat69781aggatattga accctcatga acccactacc aaacttcaac aattactaac tcttggttat69841gcttgtttca cccatgtgcc ccacctcctc ccttccccct tttattttga agcaagtcta69901agacactgta tcatgtgatg gacattaata gatacttgga tagagtactc tgagaataca69961ctgggaaaac atactgtatc gggacaggga aaggagctac agaaaagtaa catttgagtt70021caatctaggt ggggaggagg atgttgttcc aggtgatggg atggaaactg acaattatta70081caatgtcttg aaaaatgtgt atatacaact tttttgttcc tctgaactga gcacccaaga70141tgtaatttcc aagattagga aattgttttg cctgaggact ttataaaaat aattaattca70201cttttaattt agataaacta gaataatcca tgagtagtgg taaYgactaa tctgttctct70261tttcacccca gtgactttct agactttgaa cattagaatc cactccctga aaaagtcact70321ctcctttgta actagagcaa cagctatcct ttgtcacctt tatttttctc tttggaagag70381gaggaataat cagtggctca cttctgaaac atcatgaata ggtagaattt agtaatctga70441attaaaaacc tccctgtaaa ctgaaggaag aggaaattga ccagtgggca cacagaggtt70501ctcaagggac ttttttggtt agccactagg tgaaatcctt gatatttcat catgtacctt70561gtctcatttt cctgttgtga aattgggtgt tcagcattgt ttttcttgtc tgttctcaga70621tcacagaact gtgtggtgca aagcgggttg gttattttgg tccaacacag ttttacattg70681ccctgaaatt aattgctgca gcacaatctg gcctcccggt acggatagag agtattaaat70741gtggtgagta tctcacattt gtatgtttta ttgtccatgg cgagtcgctc atttatgaga70801accattttta atgagtttaa acttttgtgg aatttctcct tgcttcctcc aactcttctg70861caaaataaaa taagaaaact tggcaggtag gtagcttagt ttctttcttt tttaaatggg70921gaaaattctg agagtaacct gagattagtt cattttaact gatttgcagt taaaatagat70981tttattcttt agcaggaaga gattgggcca ggaattctag agcctgtggc aaataatcac71041attgacttta attttccact tgaggtatct ataatgttat tgaaggctct gttagaggag71101taacaaggga gtttgagtgg aactatgcct tttctctttc ccatacctgc cagtgctcag71161cctttaattc ttagtttcca aagcactgtt ctgatgctag taagtacttg ggtgctacca71221aaattttggc agaatgaatt tatcttctta aggttctgat ttaatatttt ctcatcttgc71281catccctgcc ctcaacccac tgtatgcttg agcactttct acaatttaaa caaaaagcaa71341aacccccaac tattagcagt atttctgctg cctcaggatt ggttttaaaa atctgtatga71401aaaaggtgat tattcaaggg gtgtgccaag gtcctctgat aattcagttt gatacagatt71461actttcatat gtccatacag gtatgtagga actttttttc tttttctcat tctgagacta71521aaaacctctt aactgctcat gcctgtcctg tgaagtctct gatagacata caagtggatt71581caggatttat ggtgagccca tggaaacctc gtaggacaat ccaaatggcc gagaagattg71641ccatagatgg ctggctgtgg aagaccaagc ccttcacact aaccgtYagc aggtggcagt71701catcatgcta atctttatca gggggctagc ttgtgccaga cactttcatt taatcctcat71761gacaccctac taaattaggt acttgtgtca gtttgattgc ttaggataag ctgcgataac71821aaatgtcaca atctcagtgc ctcaaccaga cagaggttcg ttttccactt tgcagtgggt71881cagggtgacc ctccagggca gctgccctct atgtggtgac catgtgatcc tggctgcttt71941atcttgtggc acctcccaac acaacacctg ttcccataat cgccacacca gggaaagagt72001atctcaccct agcggttaaa tgctttggcc cagacatgac tgaagttatt gctgcccaca72061ctgcattggc cagaacttgt cacattaatc tgcttcagtg cagagcaggt gggagaacac72121aattctctcc tgtgcctgca aggagaggac aactgcgagt ctctactatg gctgcacact72181gcaagtctct actatggtta tggctgcaca ctgcaggtct ctactatggt gctgccgttg72241ttcgtatctt acaagtatgg aaatttgaac tttagggagg ttgagtaact tacctaagaa72301tataaggtta gtagaatttg gagccaggat ctacttccag tccccatgct cttaatttct72361atgcaaaata gccttgtctc aattttttct atttagtaag atgctgacag taatctgtta72421tacaatattt tctgaatggg aaaaatattg gacttgaaag gcatctcttg gtatgacaga72481attttgttct ttgaaatagg tttcttgatg gagtgtcatt tgacaggcag gcttagtatt72541cattgctgca agacacagcg gccatttgct ttgattagtt gaaagttgca tcacactgaa72601tttggatttg tggtaggcac tgatagtttt tattccattg taaatgatca ttaagcttta72661ttatagtatc ttccaatttt atcttctgga aaagttagag ctgctttatt gtatacagaa72721taaacttttg acacttttgt tacttatttt aagttagacc taagtctttt acctgatttg72781tgcccagatt gacagtttaa tttcctgagg gcagggagca ggattgttga agttatcact72841tgtgtaaagg attaacagtt tatcacaaat ataaggcttt ctcagtttta ttttataaac72901ccagttgctg ccttttacct ctgagagttt atatatgtcc agagttaatt ctgaattatc72961ttccttcacc actctctact aatccattta aagattgatt aaataaaatt atttgcagtt73021agcatatgtt gaataagaat tagggcaaga ttgggccaaa ctttgagctg ccacttagaa73081ttttccagtt tgttgcttga aatctattag cttaatgctt gatgacacaa atcttgaagg73141ccaaagggat cactctgtga gaacctaatc tgaaaggctg tgctagccct gtcacaccca73201gggcatcata cccagggagt tgtggatgga tacccttatg tcatgaataa catgcacctg73261tgtgtatgtg cgtgaccctc taatccctag tccttcccat ttcggtaaat gccaccacca73321tcctcagctg cacaagccag aaactcagca gccatcctgg attcctcctt ttccctcact73381ccttaataga atccatcaaa agtcctagca atgtggctcc aatctcactc cctgttgcct73441ctatctcttt gtccctgcta gaccagacaa ccttcatcta cttacctgga caccacaatg73501gattgacatc ctagcccctc ctctcttctc tactgtgatc tctcttttcc cactggtttt73561ttgtttgttt Nttttttgac agagactcgc tctgtcaccc aggctggagt gcagtggcac73621gatcacagct cactgcagcc ttgacttcct gggtgcaagg gatcctccaa cctcagcgcc73681ccgagtagct gggactacag gcatgcgcca ccactcccta ctaatcctac taatttttgt73741attttttgta gagatggggt tttgccatgt tgcccaggct ggtcttgaac tactgagctc73801aagtgattgg cctgcctcag cagcctccca actttttttt tttgagacag ggcctcactc73861tgtcacccag gctggagtgc agtggtgtga tcacggcttg ttacagcctc tgcttcccag73921gctcaaatga tcttcctgcg tagctgagac tacaggcata catcaccatg cctggctagt73981atatatgtct ttttgtagag atgggggtct tgttatgttt cccaggctgg tctcaaactg74041ctgagctcaa gtgactctcc tgtgtcagcc tcccaaagtg ctgggattat aggaatgagc74101caccataccc agcctctttt cctacttttg aaagtgtgat cttttcaaaa tacacagcag74161attattccat cctcatcctt taaaccctcc agtagtctgc ttgttgtact ttgtaaaaca74221ttcaaagtcc ctacactgat ctgtaacacc ctacacaatc tagttcttgc ctacttttcc74281atcctcatct caaatcttaa acccttgctc actacctgtt ccccagtctc agtggccttc74341tagccatttc tgggtgactt ttcctgctgc aggccatatg tccattttat cttgcagtct74401tgggttccta gggatctgtt tttcctttct gcttcccctg gttggctcct tcccaacctt74461caggattcag agtctgtccc tactatccta cctaaagtag ttctgtcaga ttcattaatt74521catgaaggaa ccagcatatg gtaaaactgg ctcaagaaag tatagagagt cagatttatt74581tatagtcata gaaagaaagg atgctggaat agaattgcta aattaggtat aaagctaatt74641aacgccctac ggggtaatac aagtgcagcc tggctattgt acttttttct aagatgaaag74701tatttgcatt gccgtcagac aagaattact tgttagctaa cagttttttc caagttaata74761tctacccttt caggattatc aaatgcccaa ccaagagtaa acagtaactc agagcagaag74821tccatatccc taggaagtga ggtgatcctt gggtgggcct gctgggtaat gcttctgcag74881ggcaatctcc tccccacttt gttctttccc agcatctgtt tcctgatagc acttagcaga74941gtttgcaaat attccttaca ggtttcacta ttttttttgt ctgtcttcct cacttgtcca75001gtttgtgtaa gggggaggga agaggatctg acaccccttg cattgtgggg cattataggc75061actctgtgtg tattggttga atgaatgaat gaagaggtgc atgtctacgc atgaggaggg75121tgtcttaatc atctgccact ctttaatcta aaacgattgg aattcagatt ctcctggaac75181tccagaggag tttttacaga agtcatctca gtctgcatat catctttctt aggcactgat75241tttcaacctt gtcggcactt gagaaccacc tggagagctt taaatataaa ataaaaaaat75301acagatgcct gggtccatct ccagaaattc taatttcata tatctgcagt ccggtctgag75361catcagaatt tttaacagct ctccaggtga tttttgtatg gagacatgtt taagaataaa75421agctgcatca agcattttta agagctttgc ttaaactgtg tgtgtgtgtg tgtgtgtgtg75481tgtgtgtgtg tgtgtgtgta gttacagtag ctgcatctga actagggtgc tgtacgacac75541agggcacatg atttaagggg gcacttactc tcagggcaag tgccgactct acatttacac75601caccctgaga gtgcgtgcct ccttaaatgt tggcactagg tatctctcta gccccaccct75661cgtctgagtt ggggttaaac agatggcaaa acagctaggg cagaaggaaa gaaggtgaaa75721attaaggact ggtagagaat ttctgatgag gaaatcatgt ggtgaacatt tgccctggga75781cctgctgtta cctttctttc cagggtctga agttgagcac gtgtgaggac tgagggaagg75841ttaggcagaa cctagagtag ggtttctcat catttttgtg ccaggagccc ctttggcagt75901ctgaagcctg tgggcccttt ctcagaatta tgttttaaat gcaacaaaga cataagatta75961taatggaaac caattatact gaaataatca ctaaaatagt taaattgtta tataacaaca76021tacatgcttc taggcctggc acagtttctc atgcctgtaa tcccagcact ttgggaggcc76081gaggtgggcg gatcacttga ggccaggagt tcgggaccag cccggcaaat gtggtgaaac76141cttgtctcta ctaagttcga gaccagcctg gccagcatgg tgaaaccttg tctctactaa76201aaatacaaaa attagccggg tgtagtggca catgcctgta atccctgcta ctcaggaggc76261tgaggcagga gactagctta aacccaggag gtggaggttg cagtgagtcg agattgtgcc76321actgcactcc agcctgggca acagagtgag actatatctc aaaaaaaaaa aaaaaaacca76381acccccaaaa catatatgct tctttattaa tgcattaaat aaagcatcta acagtgtttg76441aaataaccac tactttttca aattagtaat atgcataaac agtacttcaa gatgttggcc76501cggcatggtg gctcatgcct gtaatcctag cactttggga ggccgaggag ggtggatcac76561gaggtcaaga gattgagacc atcctggcca acatggtgaa accccgtctc tactaaaata76621gaaaaaaatt agctgggcgt ggtgacacac acctgtagtc ccagctactc aggaggctga76681ggcaggagaa tcacttgaac ccgggaggcg gaggttgcag tgagccaaga tcacgccatt76741gcactctagc ctgggcgaca gagcgagatg ccatctcaaa acaacaacaa caacaacaac76801aacaacaaca acaaacaaaa ccctaatgtg ataggtgaca aagttaccgg tattgctagt76861gataatacag tggcttgttg cctacattca taatggaaag aaatgctaaa tttcagttat76921agtagggaaa atgaagttgt aatttttttt tttttcctaa tcaaggtgac agaccccgtg76981atttcttttc actagtccct ttgggagcct gtgaactctg atggtgcaat gacatgattc77041atgtagtctt tggggctgtg ttccccaaac tggtttatgg tctggttgca gtctggcaca77101gtttagaatt aggacaaaag gatgagttat ttataaagct aaatgtattc aaatctaaga77161ttgggttttt tttggttcct cttgttaagt gtcctttgtg aaactgctaa tgggtaatag77221tttggggtaa aattcttgac gagataaaac attggcaact ttatgttggc tttcttcatt77281tctttttttg ttgttgttgg gggagagatt tttcctattt tgtgaaatcc ccaaatttgt77341gagccactgt tgtggtcacg attctactgc caagaagata ctgaggcaaa tcattgaatt77401ccctttgact cctcccttat ctgcaaaaca gttgttctcc acctaggtga tttcgtccct77461ttagggactt tcggcaaggt gaggagacat ttttggttgt cccagctgcg gaaggggact77521gcctggcatc tagccggtag aggccaggga tgctgctcaa caccctacac tgcagaggac77581aggaccccct cttccaacaa caacaaactt atctaatcta aaatgccagt aatgttgaga77641ctgagaaagc cttctgtaaa ataaggatca agacttagga actctaatcc tgctgccttg77701cttcctgtgc tctagttagt ttttttgttt ttttaatcat acaagaaagg tagaaggatt77761ttgcagcatt gagtcctatt tctgtggctt ttgttttcaa agaagaatta ttttattcaa77821ggaaaaataa aagcagttat tctctggttg aattgagggt ggggttccct gtgctttgcc77881tatttgacaa accctcccta tctcttaggg ctaggagaga aatcttttga gtgtgacaac77941cagtttggta attcatagaa aagacaggat aggaagccca agggtaaaga aagaagccca78001ggaggtaagg ttgtagctgg ctctgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt78061gtgtccacac agtgtctctg tattttgaac actaaatact atgttgggat gtctcattaa78121gctaaggtca actcatcctt tctgactgaY gtccaaccca atcccccttt cctttttttt78181cctcagtgtt cctttgtcaa tttcctttac cccctctccc agccctgcta cagggacaca78241gagctgcttt ggacaaagtt cgcctatgca ggttcactga ctgtacggct ggacatttgt78301gttttctagg ggtctgtccc tctaccccat agcgctgtca ttagctctat tgttgctcct78361ccccaaaggt tctccccaac cttttgagct gtggattttc accttatttg taaaaatctt78421tgttgtggga ttctttgaca atctagtaaa agctctggat cttagcagga aaatacacat78481tgacattatg taaaaaatgt tgccttcaat gtcaggggtt tctggacccc tagagctggt78541tcatggagcc cagtgaagag cccttcttcc agagaaatgc agtgtatggg atggatatag78601aggaggtaag ttggaagatg catagtgggg aaattgaagg ccaaagaggt aaaataaaag78661ggatgctgct tgctagcagc agagcccagt ggattgtgga cctcctaact cactcctggg78721gttctacccg tggtatggcc aggaaattga acttatccac catcctcctc tgccaaacat78781ttacaagatc acccttccca ttgtaacctc caccccgcca tctagcgatt acctacccat78841ttcctaattg acaagaggta tttcagatca atgggcagat aggaggataa taggagacta78901gatagctttg attactcagg cctgggttga taagactggc ctgatttttt tttttgtttt78961gttttttgtt ttcccccaaa gtaacttcag gaaaatgtgt agggatggtt tcactttgga79021attatagaac attattgtca taattctttg ttaacgtata agtagctggt ataagtgaat79081aggtaacaaa aagcagtggg agagaaggga gccagcagtc aggagtcaag aatttttgtt79141acggtcttag ctctgccact gtattgctgc atgactaagg caagtcttgc tctctctttc79201ctgggcttat ttcctcacct gcaaaatgca gacatttgag caggttttct ctagattatt79261ttctgaaatg ccacagtttc atgctaaatg ttcaaatctg tactagaaag aattttaatg79321ttcaggatgg tagtatagcc ttgtggctgg gaatgtaggt cctagcaata ggctacctca79381ttatgaatct ttccttcatt accttttatt tgtgtggcct ccggccagtt tcttaatttc79441tctgtttcaa atgactccct ggaggaaaag gtataccaat aacacctgcc ttatagactt79501gttgtaagca caaaatcagg gaatccctta tcaaacattg tagaaaagca tctacaactt79561tgttatttaa tatcgtagcc acttgccatg tgtggctatt gagcacttga agtgtggcag79621tgtggctagt gtgactgaga aactgaattt ttcattaaat gatttaaata aaaggtttta79681aacagtgtaa aatattttct gttaaatgca accttattga tttagtagga ccacttttca79741cttcaaccat caaattctgt aagacaaggg tcaacaacct ttttcttttt ttttcttctt79801ttttaaattt tattttatta ttattatact ttaagtttta gggtacatgt gcacaacgtg79861caggtttgtt acatatgtat acatgtgcca tgttggtgtg ctgcacccat taactcgtca79921tttaacatta ggtatttctc ctaacactat ccctcccccc ttcccccacc ccacaacagt79981ccccggtgtg tgatgttccc cttcctgtgt ccatgtgttc tcattgttca Rttcccacct80041atgagtgaga acatgcggtg tttggttttt tgtccttgca atagtttgcg gagaatgatg80101gtttccagct tcatccatgt ccctacaaag gacatgaact catcattttt tatggctgca80161tagtattcca tggtgtatat gtgccacatt ttcttaatcc agtctatcat tgtgggacat80221ttgggttggt tccaagtctt tgctattgtg aatagtgccg caataaacac acatgtgcat80281gtgtctttat agcagcatga tttataatcc tttgggtata tacccagtaa tgggatggct80341gggtcaaatg gtatttctag ttctagatcc ctgaggaatc gccacactga cttccacaat80401gattgaacta gtttacagtc ccaccaacag tgtaaaagtg ttcctgtttc tccacatcct80461ctccagcatc tgttgtttcc tgactttttg atgatcgcca ttctaactgg tgtgagatgg80521tgtctctctg tggttttgat ttgcatttct ctgatggcca gtgatgatga gcattttttc80581atgtgttttt tgactgcgta aatgtcttct tttgagaagt gtctgttcat atccttcgcc80641cactttaaag ggtcagatag actttgtggg cagtatgatt ttggtcacaa ctactcaacc80701ctgttgtagt atgaaagtag ccatggacag catgtaatca gaggatcatg gctgtgtgct80761aataaaactt tatttatgaa cactgaaatt tgaattttta atttttttta ttttttattt80821ttgagacaga gtctcactct tgtcacccag gctggagtgc tgcagaggca cgatctcagc80881tcactgccac ctccaccttc tgtgttcaag ctattctcct gcttcagcct cctgagtagc80941tgggactaca ggcctgtgcc accacgcctg gctaattttt gtattttagt agagatgggg81001tttcaccatg ttggtcaggc tagtctcaaa ctcctgacct cgaatgatcc acccatcttg81061gtctcccaaa gtgctgggat tacaggtgtg agccactatg cctggcctga catttgaatt81121taatataatt ttcacatcat gaaatatttt tcttttttct tttttttttt tttttttttt81181gcaaatattc aaacattgtg cttgcgggat gtacagaaat aggcagcaga ctggatttgg81241ttcttaggta tacacccaag ataaataaat tcatgtccat caaaaaacat ttataagaat81301ttcacagcag ctttatttac aatgggccca cactggaaac aacccaaatg tttatcacag81361gataatggat taaaattttc aatatattct tttagggtct tgctctgtca cctagactgg81421agtgtaatgg tgctgtttca gttcactgta acctccacct tccaagctca agccatcctc81481ccacctcagc ctcctgagta gctgggacta taggtgtaca ccaccatgcc tggctaattt81541ttgtattttt tgtagagatg gggttttgcc atgttgccca cactggtctc gaacttctga81601gctcaagaaa tctacctacc tcggcctccc aaagtgctgg gatcacaggt gtgagctact81661gctcctggct caatatattc ttaaaagtgg attgctccat agcaatgaaa aagaatgaac81721tatgtgtaca atacattggg tgattatcac agacaaaaag gtcgtaaaaa aagcttgtca81781caagaacaaa tactgtatga ttcaatgtat gaagttccat aatggcaaag ataatctgca81841agtatagaaa ttagtacaga ggttgctcgg tggaacaaag ggaggggatt actggaaatg81901aacatgaagg aaaattctgt atcttaattt gggaatttgg gtgttggtta cagggatata81961tatatatata tatgtatttg tcaaaattct tcaagtggaa cacttacaat tccctataaa82021tcatcccacc atacagaata ctctttacat aatttaaaaa gaaatcaatt agttgcttac82081atttaacatt gtaataataa taaaatacag taaaatacag atttctggcc acaatggctc82141acattcccat gtagcaacta tcaactagag ctgtgctgtt tcagtatggt agccagtagc82201tgcatgtggc tgttgagcac ataaaatgtg acaagcacaa attgaggtat gcaaggaggg82261taaaatatac atgggagttt gaaggcttaa tatgaaaaaa aatgtaaaat atttcattaa82321tagtttttaa tgttgattac atgttgaaat gataatattg ggttaaatta gattattaaa82381attttgccat taaaaatgtt ttgaaatgga aaatttttaa ttgtgataaa atacaaataa82441catttcccat cttaaccatt ttaaatgtat aattcagtaa tgtgtaaata tagtcacatt82501gttgggtaac tagtttctag aaccttttca cctcacctta caaaatggaa actctgtatt82561cattaaacac cttcccattt tctttttcct acccagatcc tggcaactgc cttttttttt82621tttttttttt ttttgagtgc agtgacgtga tcatggctta ctgctacctc tgcctcccag82681gctcaagaga ttcttgtgcc tcagcctccc gagtagctgg gattacaggc gcatgcacca82741ctacacccag ctaatttttg tatttttagt agagatgggg ttttgtcatg ttggccaggc82801tagtcttgaa ctcctggcct caagtgatct gcccacctca gcctcccaaa gtgctgggat82861tacaagtgtg agccactgca cctggcctgc ccatctactt tctggttYta tgagtttggc82921tactcttgat gcctcatata agtgaaacca cacagtacct gtctttttgt gaccagcttg82981tttcacttct ttctttcttt cttttttttt ttttttaatg tggctcacat tcttttttgg83041cagataacat tgaaagactg tgaacagctg ctgccatttt gatggaacat gagttctctg83101cccagcaccc tccactccat attgtctagc taatgaggct caatgtcact tgccatttat83161tattgcattt atgctgttta cattttttct aatggattga ggaaattgag gtattttttt83221ttcttttttg agtgacaggg tttcactgtc acccaggctg gagtgcagtg gcatgatgca83281gccttgaact ctcagactca agcgatcttc ctgcctcagc ctcctgagta gctgggacta83341caggcatatg ccaccatgcc cagctgattt tttattttta tttttttaag agaaggggtc83401ttactgtgtt gtccaggctg gtctcaaact gctgggctca agtgatcctc ccacctcggc83461ctcccaaagt gctgggatta caggcatgag ccaccactcc cagcctaaag tattcctaat83521atatctgtat ctctatcaaa tatggaaaaa caaaggattt ttctctcttg tagagggtca83581agtgtttaaa aaggagcagg cataattttc tttgtggaag ggaaaaatag tcatttatgt83641ttaatataca aagcatttct gtgttcaaag aatgtaacct aagatgaaac aacaatgaag83701caaattgtca tcttggatgg ccactctcat tgtctcctca gaccctctaa acatttagtg83761tgaactctta ctttgtttgc tgtgtgtaat tcaattcagt ttgggagaca agattctaac83821caatgactac aaatagacct tcttcttgac agacatcatt gtatagcagg tactttattg83881attaattttt tcaaagtttc actattaaat tttgtgtatg tgtgactagg aaaaacattc83941agttgagtat ggtgactcat gcctgtaatc ccagcacttt aggaggccaa gacgggagga84001tcgcttgagc tcaggaattt gagacctgtc tgggcaatat agcaagaccc catctttacg84061aaaaataaaa aaagaaatta gccagcatgg tggcaagcac ctggagtccc agctacttgg84121gaggctgagg caggaggatc agttgagtct gggagttcaa ggctgcaatg agctatgtta84181gtgccactgc acttcagtct gagagacaga ccttgtctca aaaaaacaaa aaccaaaaaa84241aacattcaga ctgaaaattt acaaactact tttacctttc actttttaaa agcttgcatc84301agtttaaggg aaatggcacc ttcatgaaag ttttccatga taagaatatt attatgtatg84361gttatacttc tgttaagtga gagttgcttg gaaccacctg accaaagaaa gtatgagaga84421acttcagctg tattttgtct tttgctgttt ggtggctgtc tgggggagag aggtggtttt84481ctaggcacat agctctaaag aaatgccata gacagggtgg aagaatgtgt gaagtgacaa84541tgtcctagac aatatctggg agccctactt aagtcctaat ataataattg atattcataa84601ttcagttagt gtattccatt gttccttgga ggtatctgtt gaatgccagt aaggcactgg84661tcagggagct tataaatact gagcatttat ttcttttctc cctcaagaag tacttggaag84721tagagtcatc ctcttatggg tagatcaggg tgcaaattat tttatgtgat cttgtttcat84781ctttttagat cctttttcct gactgagctg gagcacatac ctgagaccaa aaccatagag84841ttagcttata tttgatttac taagaatcta gagtttcttt tctcttgctt ttttgctttt84901gaagcccatt cctgctttga gtgctggaga gctgcccgag attgggaaac attaagattg84961gttactctat ttatgtctct tgggcccttg gctgattgat cacacaggga tttgtcatat85021tgcctgcttc tttttttttt ttctggtaca gaattttgct cttgttgccc aggctggagt85081gcaatggcat gatcttggct cactgcaacc tccgcctcct gggtttaagc aattctcctg85141cctcagcctc ctgagtagct gggattacag gcgcctgacg ccacgcctgg ctaatttttg85201gtatttttgt gtagagaYgg ggttttacca tgttggccag gttagtttcg aactcctgac85261ctcaggagRt ccacccgcct ccgcctccca aagtgctgga attataggcg taagtgagcc85321actgcgcccg gccgcctgct gctttgaatc aaatgtgaac atagcaacta aaagtaaaca85381tcatgtggat gttagagaac caggggcctt ctttgcctgg ggaactctag gatacttgct85441tgctgttgct aagatgtttt ctggccttag cacagcatca gtcattgaaa gtgcagctgt85501gtttttggaa tagagagctg atggaagtgc aaaggcacac tgatgagtat tatcctgaac85561aatggataca gatgcaaatc attatgcata atccataatg tccccatttt tgtagggttt85621tgtgctacgt gattgtgtgt gatttgatgt gctgagaact agaagtgtgc caaagtatca85681tttccgcttt cagaatagtt ttgttagaat tttgaaagat gagtaaatat accagaccgt85741tgcttttcaa ctctttgtag cattttgagt ggctaatggt tttgacagaa gaatgtggct85801gaatgataga ttgattgctc tgacttgata ttgttttcct tattccatta tgaaaagaca85861accagagatt ggatgtgaaa attccttaaa gaattacata ggatgagaaa ctgttctacc85921cactaagttt gccataggaa caagggacca ttctgtagag cactggtaaa gtaccagctt85981gcctgcattc aaaccgtggc tctgccacag atgagttgtg tgatcctggg gacattactt86041agctccttta tgactgtttc tacttttgca atgagggggt ggtaatgata gttttatcat86101acagagttgt tgtgaggatt aaatgagtta atatttgcaa atctttaata ctgtgcctgg86161catatttaag cattatatga attttacctg ccaccatttg tcacttatta ttgtattata86221cagtgtcata tcctttttct tggttattga aggatgaaat gggagatcag tcccaaatct86281gaatttattc tgcgatttgg caacattgaa aaatgttaag ccagaatact ttggttaaac86341catagcattt taaaaatcag gacgttatta taggagtact cattgtatct aaatctaaat86401ttatatttag tgctctggaa tttttcctca aaaatgattc atcgtttggc cattgagttc86461cccttttttg cagtttttaa attaagtctg actagagctt ctctgatttc tggtcccaaa86521gaattgcctc tgcctcgctt tatgatgtca aagaatgatg gtgagatacg atttgggaac86581ccagctgagc tgcatggaac taaggttcag attccatatt taactacaga aaaaaattcc86641ttcaaaagaa tggacgatga ggataaacag gtaaacagtt tgttcagatg tgttcatttg86701attctgacca tgaaaccgca tgtagcaggg cctcgtatct actgttcagc ttccagcaaa86761tctcaccatc tggaacccaa ttacagactt ggaagtgagc cagtaaagtt ttctcctagc86821aagaaagaga gaccgacaca tatgaatcac caccacccac ttaagggccc tttagagcct86881actttaaagt agaattgtgt ctaaaataag gctaccgatt taatacattt attgatttca86941ctagctcagt ggttttctaa atggattctt ggaaatgtaa cttccaggag tttttaatag87001gttctcttct ccaaaaagaa gtctgtagaa aaggtttctc ctattataat aaatgtgttt87061ggagaatgct gcttcatatt cccttgttga gggaaggact aaatcctcag tcttggaaat87121tcgtagtgca cattagcacg gtaaaagccc tgagaagttc tgcagtgaag agacctggaa87181atgtgtgttt actaaacttc tttcaccagg aaattctttc ttggggaaca tatttggaat87241actatcttgg gaaatgctgc cattacccat ttagcaggtt tggaaccaga acagggagga87301taggctttta aaattagacc actaattgca atggtcagat gatgattggt ggagacaaca87361gtataaaatc tcagaagagt gggcagatcc agaagaagta aatagtctac acattRgtgt87421accctgcagg catcaaggta tagctcatac ttccagcaaa gatactgaat catgcagaga87481agtcaaatta tgttcaacat agtgtcttct ataaggtaaa gaaatttatt ataggaacac87541ttgaggctgg gcacggtggc tcacgcctgt aatcccagca ctttgggagg ccaaggcggg87601tggatcacaa gaggtcagga gttcaagagc agcctggcca acatggcaaa accctgtctc87661tactagaaat acaaaaatta gccgggcgtg gtggcatgtg cctgtagtcc cagctacttg87721gaaggctgag gcaggagaat ggcttgaatc cgggaggcgg agattgcagt gagccgagat87781cacatcactg cactccagcc tgggccacag agcaagactg tgtctcaaaa aaaaaaaaaa87841aggaacactc gaatgaaaat gtttttaaaa tgcaatgaat attaaatgat tatgttggaa87901aactaacttc tgtgacttac cctgtgatgt tgtgtagatg aggtgttgct acagaagttt87961tatatggata gttaggggag gtgtagcaca tcctgttaaa caattccatg tactgtcact88021ttcataattc tgacacattt taaatcagtt cttcagggat atatttaaaa aggaagaggt88081ataggaattt tcagtcaata ttctttggca tattttccca ccagcatttg gagaggaatg88141agggtccact gtcccacggt ggttgatgcc ctgggaaatt tccaggccca gtccctgatt88201gtgtgaatga gaatacatcc tttcacttca ggctgagttt atgcatgccc cacccaatct88261atgggtgtga gctcatgctt tagagcaaac atattttgct ttttcttgaa gaagagtatt88321tgatcagtat ttcagattac atatttctct ttagactaag ggagcctata tatatagtct88381gtgtgttgat agaaagcatt gttttggctg ggtgtagtgg ctcatgcctg taatcccagc88441agttggggag gccgaagcag gaggattgct tgagcccaag aattcaagac cagcctgggc88501aacatagcaa gacctcattt caaaaaaaaa aaaaaaaaat tagctgtgtg tggtggcgca88561tgcctgtagt agctactggg ggtgctgagg tgggaggatt gcttgagccc aggaagtcaa88621ggctgcagtg agctatgatc acgccactgt gccccagcct gggtgacaga gcgagaccct88681gtctcaaaca aacaaacaaa caaaaaaaag cactgttttc ttggtacttg aatactccaa88741agttaccagt ctactagtaa agtttttttt tttttttttt ttttttaaag aggaaactca88801gcagcagata aggttgctta ctttgcctag ggttctacat caggttcata tttgaattga88861gattcaaatt gtagcttgtt aagatttcat cttagtgttt gggagtaagc tggttggaat88921ggtttaccta taatgaattg ttttgcagtc tctcattgtt aaaaaaaaaa agctagcatt88981ttactttgac atggtaagct ccaagaatat taagtattaa gcatttagaa atatatattt89041taatattcac atttgctgga agacttgata agtataaatc caggaaataa atttattaat89101gaatgaaaat gaaatctatc cacagagaga tcacaattgg gagcgggtat gaatgttatt89161tcatgtgatc agtgtctttc attttccaaa ggaaaatatt attggaataa ccaaaagtgt89221ttcaggctgg ttttagtaat ttaggtaact cagcactgat gagattttat tctctgggag89281acttYttgaa tcctcagaca ctgctgttta accatgctgc atttttgaga aaacatttcc89341ccccatttgt taaaccagct gggtcaccag cagtagagtt gcgttatgct tcctgactgc89401agctcagaac gccaggcttg agtgagcgcc tctgaactct gatctggttc tttgaagcag89461gaaacacagt ctcccacgat gtcacccctc gcctcccctc cttcttcccc gcctcattac89521cagagggtgc ccttgagcca tggctacagc aaactgcgga gcagcgcaga acagatgcat89581ccaggtaaga ggcgacctgg gggccagctg tcccaggcag tgtcttagag actccttaat89641ttaYcaggcg gtaacgcttc aagctgctta atttcctgat ttaattaccc ctgtctttgg89701aaacatggct caatttaatt ttatattctc aagcaaggtt tttttttaaa aattgattta89761aacatatttg agtaatgttt catctcctgt cctagaaact agttcgtctt tagacttgta89821aacagctaga tgatttaatc tttcatatac agattttatt gaatagaaca taccatatta89881agggcttgta ttttttttgg aattgttatt cctttctgta agcacaccaa agatatttat89941atagtaactg ctttaatgaa atttcttaag aaatataact gcaaaagcaa aagagttgat90001ccaacctttg aagactgagc acgtaaatga aaatatatag gtcctgtgct ttggaataag90061gataaaggaa atgctagggg tagcattttg ctgaggggta tgtagcatgc acagttgtgg90121ggttgccttc ttgctcacat ttatgctctg ctggcctcgt taggtggtaa aatacgttag90181ttctgaggat gacttgcaga gttggctgtt taggctgcag tgcctaaagc actggaaatt90241cactgcagca tcttgtgcct cctggggttc agactcctag tttaagatat gaccaattaa90301aggtattcta gaactggata tctgtaaacc cccacccccc cagttttgtt ttgtgggctg90361tggccccaaa tgacccagtt tctgcaggtt tttactgcat taaaaatagt ggtgactttt90421agagggacac agaaataaaa ctgcgcgcta ggctgaataa agagaggctg taatcttgca90481gagagaagca ggcgtggcat aatggcaata gcttttgatt ggaagcagaa ggtgtgtggg90541tttctgtgac tagctgtagg actttgggca ggccattgaa catctctgaa ccacgattac90601ctcttctctc ttaaaatgga gtaacagtat cctagagggg tattagaaga ttaagtgaga90661atgtgagtat aaaatctgtt tggaaattaa gtcctatgca catgcaagat gatgttgcta90721atggatagta acaaaaaaat cactgtcctg taacttgaat atctcagtgg ggttagaaac90781taaatggagt tgagaattct tcattttcct cttatagttt ccctgatctg atttccctat90841tttcaggagt gggttttttt tttttttttc ctatttctaa tcttacttgg gtttggaagt90901tggtttttaa ttcacaggtg aaactttttg ttctactgag tcagcctgtc agcaagttct90961acctttggta ttccttagaa aacttttctg ttcctcctcc ttaatgcccc attccctcct91021cattccatac aaattgggct tccatttccc cagtggtaag gaagcagctt gcttctctcc91081atctttcttg actctcagca gcctgtgcca aagctgtcca tgccttcatt tcttttcata91141cttttttaaa agtgaggtaa aatatacata tgataaaatg tacccgtttt atttgttcat91201ttttgagaca ggttctcact ttgtcactca ggctggagtg cactggcgcg atcatggctc91261actgtagcct tgatctccta ggtttaggtg atcctcccac ctcagcctcc tgagttggtg91321ggattacaga catgtgccac cacaccgggc taattttctt tttgtatttt tttgtagaga91381tgggatttcg ccatgttgcc caggctggtc tcgaacccct ggactcaagc gatccgctct91441ccttgtcctc ccaaagtact gggattacag gtgtgagcca caatgcctgg ccatgcacat91501gttttaaatc ttcaatttga gttttagcaa atggataggc ccttgtagct accactttga91561tcagtaattg aacatatcca ttatgccaga gatgaccctc tccggtcact ccctgccccc91621taacgcaatc actgttctga ttttcatcac catcattagt tttgcttctt ctagaacttc91681atatgaatga aaacacacag aatgactgac tgtactcata tgtgtttgcc atctttcctt91741cagcataatg gttgaaattt atctgtattg ttgtatcagt agttcattcc tttttattat91801ttagtagtgg gatatattgt gtgaatgcat cacagttcga cgatctgttc tcctggtgct91861ggacatttga gttgtttcca gtgtttggct attatgaata aggatactat gaacattcat91921gtatgagtcc ttctgtgaca tatgctgtca tttccctggc tcctaggaat agagtcactc91981agttttaggg tagttgtata tgaaactgcc aaagagttct ggtttgacac tgaacttcag92041agtgatgaaa tctttaggtt tttttttttt tttttttttt tgtacacctc cccttggaga92101aagatggtac ttccagttct tattaaacct cttgtaaggg gatcaaagaa ttatttgttt92161tgcactgtat tgcatgtttg gggtttcatt tgggaagaaa gagagttgag taatagctac92221tttttaacct tctgaattca gatccaatta aggaaggagc cagaagtcag acaattgatg92281gcctgaatat cagctttatt actgatatag ctgatcagag cacatgtctt agatttgtga92341ccagatgggc tttagacact cacctttcca gtcacaggca ggcttggcca tcccagattt92401cccacacaag taggaagaga aaatctgcct ctccaaaaaa ggccccccat gcagctgagt92461cttcatccaa gccctactga tgagataggt ctgtctcttc caagggaagg cccttgtagg92521tgagggatag aagccaaggg ccagatggtg gcctgttccc ccatgtgtct cccctggaag92581tagaagtaga agctcttatg ggaagcagag gcacgtggga ttcacttcct aaggaaagct92641catgacYgct agcaccccaa ccatgccaca gttcactgtg tttcaaacct tttggcagag92701cgcaattggt attagtgatt taattcagtg ctccatcaac acagaatgat tttaaaaatc92761ctacttctca gaagacaatg gtggcctacc agctgagact tactttttaa aatttggatt92821atgtgctctc tatgtatgta tgttttgcag tttttatatc tgtattcatt cttagcaggt92881tctgaaaatc tacttgatgg gttgccaggc aattgttttg ctattttaat gtgtgttaat92941gtattgtaac attgtgttcc agttactagg aagaaataca aaaagcagat gtgagcccaa93001gagtgattta tcattcttga tgtgctattg ttgtaccttc tttattttaa aaagagcaaa93061ctatgttaaa catgaggtca taacaggctg ctctgctctc tacctaattt tatttgtaag93121taagctacag aaataaaaca ttttacttat ttatatattt tcaaaaactt gaactattaa93181cctatagaga tgagaaagaa tttcaacttg ggtgataatt tctgctttct gttggagctg93241catactacct ggttatgttt aactgtctcc ctgccatatt ggtaaggtca ttattaagtc93301ttctggaaat aaaaggagtg aaaacataca ttcatttttg tggcttaaaa agttatttta93361aaggcagcaa aattgttttt taaaagtttg gttattgaat agtagaataa taaatataaa93421atatatttat tcattttcgt agtcttctct acatgatttt acattctgtt tgatttattt93481ttcaaaaata atgttggttc ttagtgttca aaatgcacat tagttttctt cttggggaag93541tggatgatca atggaagaca tttacttagg ggggatatat agaatttacc tcatatttag93601aatttgaaca cttatgacac ttttctagct attgttgctt tctatcccaa ttgaagttct93661cactgttcca ccttcacaat ttattcctaa tcataaccca gtgtaaatat ttttctgctt93721atcatgaatt ggttgaatgt ctttagattg tttagtgaaa ttgtgccaaa gagaatcagg93781ggttgtcagt cctagatggc cctatgagta ttctgttgcc agttctttcc tgaaaagcag93841atgctgtatc agtttgaatg tgaagtcata atttttgaat ggaatctttg cataccatac93901tgactttcct ttctgctgtt tcagcacctt atgaagctag gcagcccctt gtccagcccg93961agggatcctc atcagggggc ccaggaacca agccccttcg gcatcaggct tcccttatcc94021gggtaagtga tgatgcaggg ttatgccaat gggacagtag agtcaagctt tgggcttcgg94081ggacatggct ttttgatgga ccctttttca ttgcaaggaa gatataaaac tgatccttca94141tgcaaatatt gcatgtcttg tttggttgct tgtataaatt actaaatctt ttctactgtg94201tatgtctcta cgccttctca gtgtctctag aataaactag gctgcttctt gatctttagt94261atatctaaaa tgtccctctg ccttcaccta tctaatcatt agccaccact ttcaggagag94321cttcactttt tctaaagcag cccatcttgt ctctgcctta ttctttatta ttactcattt94381aatgcatagt taactgtttt ccatttgttt tgtgcctcta attacatggt aaacttttgg94441aaggcagaag tagttctgta ttctctctca gtgcctagac caagagaaca gttattgtac94501aatggcagca gctgctgttt ccttatgtaa taaatacaaa agaaatatta tggcaaaagc94561ttctggctat accacagctt agacaatttc agcttccctt tctgcagtgt tttcctttgg94621ataaagagct cttattttgt tctctgtctg cagctgttgc cttgtgtagt gtctctgggt94681cctgtgctgg ggaagttata ttctcaaaga agggtgtgtg tgtgtgtgtg tgtgtgtgtg94741tgcacgcgcg ggtgtgcagg tttactattt gacactaaag aggaagtggg gcagtttgta94801taacatttca gtcatctaac tcgggaataa ggaacatttt tgcacaggca cttctactca94861gaatagaagg gtggtgtcag acactcccat aaagatgtgg gctgctaggt tggaatagga94921taaacaaagt ctaaagaagg tcctggggtt catggaatac cttctgggga tggggcactg94981tgttagatgt ggtgcagaac ttgattttct aaggtgggac tctgtgtaca aataatttct95041taaagataat aatttagcca taggctattt tttatattta atgcttccta caaccataag95101atctagttac tgctgcttct aatagtataa gtgagttgac tgtggttgag agaggtcaaa95161tagctggttt aaggtttcat tcataactag tcagtgacag agccagaatt tgaatccagg95221cctgtccaat gtcaaaacct gctcctttca ttactttgtc ctacattaga gtcccatatt95281acaagaaggc taggttctag agttgctaca taaagaaacg ttgggtgtag taaaaaatac95341tcttgaaaaa tcccttattg ataccacact ggagaaccaa tatgccattt gctcccaggg95401ccagtacagc tcagttttat ccccaagttg gaaaacacat ttgtttcttc agctgagctc95461cagatgtagt agttggctgt gtttagagat cctgtggtac aaaaatatgt gcctttaaac95521acgagaatca cacccactgt ggagtgaggt ttacgttaga aaaggtatca gtaggactgg95581gatttatccc taagctctcc aaaatgtggc tgtgtaggtt cctacagagt gtgtcctggg95641aatacagagg ccagatcctg aatggttagg atgatagtca aactaccagt gcatactgag95701ctttacagca gtgtgactgg gctcactaga ccctttctac ctagcccctg gctggcatgg95761gcatggatgc tcatttggat tggcaccctg tggcttctct tctaatctcc atgttccctt95821ttctgctgac aaacatgctt catgagccct tcactcaact aatatttagt gaacacctat95881taagtgccag gcactgtgtt gagaatacaa aaatgaggta taacactaac agtttctgcc95941ctcatggagc tactggagag atggtcatta accaaaccac cacaaagtaa atgtataatt96001atgatatggg atatcagtag gaattacact gtactttaag gatgaaaatg ggcatggggt96061atgacctagc cttgaggggc aggggctgat caaagaaggc ttctggtggg ggagcaacat96121tcgagttgga ataagggtac atgggtgtta tccaggtgac agggtgggag tgcgggagca96181aggtgacaag atggagggaa ccgcatatac aaagacattt cctatgggtt tgccatttta96241tgaacattat ctctttcagt gcccacagct atcttatgaa ataattagtg ttgttatccc96301attttcagat gtcagaacct gaggtttaga gaccttattt aatttgccca agttcacagc96361taggaagtgg cagggctgga gttcaaacac tggtcaaggt tcctcagtct ctggctcagg96421aaccagatag atggtggtgt cacctgttga gctaggaagc cctagatgaa atactggcac96481tggggaaaga tcacaggttc agtgtttgat gtcaggaact gaggttttga gatagtccag96541agaagctgtt gaagaactgg tagattacat gggtctggag cttgggagag agcttaggct96601ggacataggt atttggcagc aattggctat cagaaggtct catggtatag tgcagaaaaa96661accagacagg tgctcgccca agtccagcaa aactctgtgt gaccttgggc aagttccttc96721tcttttctgt aatacaaatg acttcgaatg gatttaacag cccattgtaa caatcagcaa96781cagtccgatt ctcttaatgt gtctgtggtt ttcatggctg gtgtgttttt ctttcctctg96841ttcttaggtt tccttcttat gcctgttaac ttggtttggc tggactccgt gttccagaaa96901gcaaaatgcc aacctggatt agtcatgtag atgaggggat ttgaagtgaa ggttcttggt96961gataagggat taatgtttac tgggcatggt gctaggccta ggctggggtg actaatcccc97021ctcatgttgt cacgttgatg tgccgatggc aaccttcttt ctcttctaac tagttctctg97081gccttgtatt caagttcttc ctgcctgatt acaagggggc atgtcctctg ggtcctgggg97141gaagccttta gggactccca gccagtgctg ttgcaaccca tggtgttggc attttagagg97201tgcttatgcc aatccagtgc ctaaacattg gaatgatttg tctttttgct tactatttta97261ggaatttcac ttttgttcgg gtgttggatt tttttccctt tgaaatttca gacactaagc97321atatcctatg ttggtagatg attgatagtg ctttgtaaag aaattataag acttattttc97381tgcttcatct gttatcatat gtttagatgt tgttttgtgg tttgtttttc acaaattacc97441ctcaacatct agatgatgta gtcatttgta attttgctgg caaaagattt agaattgcac97501gctttattcc tactgttctc agctgcctct gtttttgcca gggtcatgag aaactttatg97561ttaagccgtt gtctttattg tctccttggc agctgtggac actgaccatc ttctcgaaat97621gctctcatca tttggtgtcc attatgcctg ctctcctggt tttcctcatg cttctatggc97681tgcccctgcc tcatttgttt caatcctcct tcctctgccc tccccttgtt atgctggaga97741ctgctgggcc tacttctcac tctacttact ctttagatgt gtggatgctc atgaccacat97801agcaccatct gccagacatc ttcactctaa tgttccactg tcactttcag ctgacatgtc97861tttctcataa aatcagcttt cactcttcca ctcctagctc cacccaccca tcagcacagg97921ccagaatgct ggtcattgtc tttgacttct ctttctctcc ttatgcctgt tcagtcccca97981agtcctgacc ataccacttt tcaaatagct tgtgtctgtc catgcctctg tacttcttcg98041ttcaggccat tatccttttc actgattgta ttgtattagc ctctctcttg tttcttcttg98101cttccaccct tgccttcttc caattcatta ttctcattgc acacaaaaac aatcttaaaa98161acacaaatct aatcatcctg ctgacttgat tgaagcctgt ccctctctgc ccatggctct98221taacatgtca tccatgttgc tgtctggccc tcgcctacct cctcggccac agcagctcac98281tgccgctttc cctccactct ctggccattt cccggatggt cagtgttctc tgcgtgggag98341ccttactctt caccctcact tttgccttct ttgttttctc ctttgtctca attcatctat98401gcttttttcc ccattaagaa cgtgaaacgt aagtgttttc tgatattttc ttgtatatgc98461gatgtttcct ttttttttca ttcttaaaaa gaaaaagatc tattgcttgt cagctcattt98521atgtgtgcaa ctcagccaga gggatgggcg tctgatcatt attggtttcc catctcagag98581gcaaccaggg ttgcctgttt cttttgtatc attctaggaa cattttacgc atgtacatgc98641aaatataggt attcatatat attaatatgt gtgtgtatgc actatatgta tttatatatt98701tatttatttt tttagacgga gttttgctct tgttgcccag gctggagtac aatggcacca98761tcttggctca ctgcaacctc cacctcctgg gtccaagcga ttctcctgcc tcagcctccc98821gagtagctgg gattacagge attctccacc atgcccagct aattttgtat ttttattaga98881gacggggttt ctccatgttg gtcaggctgg tcttgaactc ctgacctcag gtgatctgcc98941cgcctcggac tcccaaagtg gtgcaatata tttttacctg aatttaggga gacatgtttg99001gcaaatgaat atattcttcc agctagattg cactagaaaa aattatcttt tatttaaaac99061caaaacacat tcacttttca aaccctcagg atactgttgt gatgttttct ttgctttttg99121gagggatgaa agtaacagtt ttctagaaca tattctcaac actcactact cttcttcaga99181gatgaaaaaa tgacaaacag aatttagata cctctattac tctaccattc ctcagggggc99241ttagctatat ttttggtgta agttttctaa tgatcaggta ggggagggga tgttaggcat99301tgttagccag caccatattc atctcgaagt gcataacttg aaaaatgttt atttaactta99361gttatctcct ttcctcctca tctcagtttc ttgccgtacc tcctccccta gatataactg99421caggtactta cggccagaag caggtgatct ccactccttg ggtcctcttg gaacccaaga99481gaaaaggcaa gtatatgaaa gatgattaaa gaaatgaata ttcctgaggc tctgtcagag99541tggtgttggc ctgagtcaaa agataattga tagtcaccac aaataaattg ttcattctaa99601gagctttgat gaaaacagca ggaaatagcc tttgggggag gctgatattg caccgagaga99661tcttcttgtg gcatgaaaga tattttatgg gtagactgga ccaggatgca agcaggataa99721ggagagcgga ggagtgagaa tgactgatgg tttgggatgg gattgagtga ggtgggttag99781agttgagtca tgagagagtt ggtcatctga gtttggaaga agagtgaagc aggtaaacaa99841gcggggcata ctgggtactg tcttggctac tctggttgtc taacctaata ctcagcccag99901ggaattgacc cacagagcag gtcaggcccg cccggattct tttggggttc aagaattatt99961taccaaggca ttacttggta agtaagtgtt catacgaacg tgtgtgagct ctcaaacaca100021cttgttcttc agagcaaata cagattttaa aaatatgtca aacattcaac agatacaaaa100081gggcacagtg aaaactgtcc ctaccatttc cccgcacagc cactcatccc aggggcaact100141gatatgaccc atttattttg tatagtacta tacaaaacat ttatataata ttgtatatga100201atatcttttt tatatacaag tgaaaatagg tattatatat aattacatgt aatatatgca100261aaacacagtt atgctttttt acacatatgg tggcatacta tacatctaac atgcatttag100321cactaacaat aagctttatt aacgttctat agcagcacat aaagtacttc ctcatctttt100381tgttttttac agatgcatag tatttccttg tctgcgtctt gtgtaaatca tttagccagc100441cccttttgat tgacatcttg tacttgcaat cactgcgttg tttgcagata tatgggagac100501caattcctag aaattgaatt attgggtgaa aatgcctata tatttgtaat ttttgataac100561gactttcaaa ttggaggtat agcagcctat gtacttgcca acaatatatg aaaatgtctg100621ttttctcgta cctttgtgaa aacaatgtgc taacaaattt atgggtcttt gccaatctga100681taggtgaaaa atgatagctc actgtagttt aattagcata tctctttatt agtgaagtga100741aatatcttaa gtttatgagt cacttgtatt tccttttctg tgaactgttg attcatatca100801agtgcccctt tttcttttga gtattggttt gtcattctta atggcatgaa accagaagct100861ggtctgtctt ttcctttgga ccagagatat actggagtgt gtgtggaaga tttctccctg100921agtggtgaga ggagatgctg gcacctgagg ctctcgaagg aggttatgga ccccagctgc100981ttcccagttc tggagaaccc caggcctgcc tgcagtgcag atttcttgtt tataaggtca101041tgacagtaaa gttgaattac ttagtttatg ggtagagatt agggctaaaa aaaaatcctc101101tgttgttaga gatggtaaat gtcaacactg tgtgtgtgcg ttgatctctc agcattgctt101161agacagctaa attatgttcc tttcactgaa atgggtggga ggaggagata gaggtagaga101221taaagataga tagagggaga gagtataagt gtgtgtgtgt gtgtgtgtta ggaagagtga101281gtttgatgcc ttttctaggt atgtatgtgc cttattcttg agatcttctt tcaccaaatt101341cattcctcgt ttcatttgag accccagctg gacgtttata ccatctccta aggggtcaaa101401ggttaacatg tcctccaggt gtcctacttc actgagtcta ctatagaacc cacatatacc101461tgaacggaca ctattgccac aaggaggtcc tattttttca tagcctccag tctcctgctt101521acttattttc ttcacctgct tgtagttctc tttctgattt tctagccctt aacttttgag101581gaaggatttc tagctcttaa cttttgggga aggaaacatt taatctcaac tgatactgaa101641gtttattttc tcagtgccaa tcttttctac ttgtttcttt tttctttttt ccttttcttt101701tcattgattg attgattgat tgattgagat ggagtctcgc tctgttgccc aggctggagt101761gcagtggcgc gatcttggct gactacaacc tctgcctccc aggctccagc aattctcctg101821cctcagcttc ctgagtagct gggactacag gcgtgcgcca ctgcgcctgg ctattttttg101881tatatatttc ttttagtaga gacggggttt caccatgttg gccagggtgg tcttgaactc101941ctgacctcaa gtgattcacc cgccttggcc tcccaaagta ctaggattac aggcgtgagc102001caccatgccc ggcctctacc tgtttcttag tgtattcctc aggtgctcac tgtgtaccac102061acttcttgac aataaaagag atgtttagtt aatttagcct cccagaggta gctgaaaaca102121ctggtttgaa ctgctggttc caggcagtac tttgtaaatc tggaaaggat agtagcttat102181tccccaggac cttttggtct cagttatatg ccaggctatg gatatggaag acttctttac102241aactggcacc attttagccc ctgggcccat cagtctttcc cttgatctca ggcRagcagg102301gagtgtcatg cccccccagt tgcctatccc aacaatagtc actactgact gaggtgcttc102361tttggtccca gtgctgtgct ggcccttttc agtcctccca acaatctcac aaggtaggta102421ttattacatc ttccatttgt ccatttggaa atacttaagt ggcttccatg agtcagccac102481catgccaggg gtacactgag gagtacacct atagcctctt gctctctagg tgcttgcagt102541caaaaggagg agatagattt gaatcaaatg atcccaccta tgcatgtgga attaaaaact102601gacaattgcc ctaaaggcct taaaacatgg taccatgtag caaaggaatc cagcctatag102661ctagaaagtg agagaaggct cctttaacaa agtatgtgtt gagctgagct gtgaaggatg102721aatagaagtt aaccaggcta aggaaggatg gtcctgggag gtgaggagag gagtggagtg102781tttcctgtag aggtaagagc ctctccaaaa ccgcacagca gagctgggct gctgagagag102841ctttgcttct gtcacaatca gaaagctaca gcattgggaa actgctgccc tagtgtcagc102901tccaaccaga tccacactac aaactacctt ggcagccaaa tgaaagcttt gttccctgcc102961tgtaacttgg gtctgcatcc gtcttccgtg ggtgtatttg ttgagtggaa cctaagtcct103021atttggaatt ctagcagcaa aggtgttttg acagtggagt ttttagctag Ycagcttgtg103081cagtgctggg acttatgctg acgggtggaa tggactcgtg acccagtctc taccatattt103141cacagggaga gcagatacat ttgaagaact gaaagaaggc agctgtggct agagcctgga103201gtgggaggga gatgaggtcg gagagggagg cagggccctg tgggctatgt gactgccttg103261caggtgggag ctgatggctc agaagttaat ttactggtta gtaagtgacc atgccaaatt103321ggaacccaag tctgtctgcc tctggaacct gtactctttt tgcacctcca gtcagtagcc103381tcattctcca aagcattctt gaattctgtc caagttgtgg atgcatctgt ttcccatctg103441aactcagact gccttgagtg tgaaggggag aacgtagtga agagacaggc agccttggcc103501tgaccagcaa ggacaggggt gggaacgtca tcgcttcgtt aatggaagtc tgcagccttg103561tcagagtctc ctcttactgc tgatttctca tttttcttgg ctttctcctc taagcagagc103621acaaaatcct gataatagtg aaggaaaagt gcccttctgg aacagcaaat cttggcccct103681gaggggagaa ttggaggcag tggttactaa tgagcatttc ttatgttaat caaaatactg103741ttagcgccac atactgcaat cacagtaaaa tgattacata cctggattgt atggctagct103801ctaggattct tttgtgatga cgtgccagat gcttacctgc aaaaattatg tgagaaataa103861aatggtgctc tgtcagtatg atttaatagt tcgccagcca aaatctaaaa ctaaactaga103921tgatggggag gaaggtgtaa tacagcatag tcatagctca ageacagate ctcaatctgc103981aaaattgcag caatgattta ggcattgcag tgaggttctt tttgtaataa atgctctatt104041tttacaggat ggatgaacag atagagataa ctatagagct tgatgatcaa gtgtacgtac104101tttggaccca gtttgtattc tggctctacc atgtgccata ggtgtgtggt cttggtaaat104161tacttccctt acctaatcct cagtttcctg ttctgcaaaa tgggaattac cggatttacc104221tcgcagggtt gtgaagatga aaattgattg tgtgcattag gtacttacag tgcctggcac104281ttagcaaagc aatgaatgct tgttacctga cattcttctg tgtctaagtg atcgtttcaa104341aaagtatagc ttattgtaat aagcagcaac tgtgtgttgc acattaggct cagagagatt104401cctacagttt ggcagtcagc cttctgaact attgaaatta gatccctaag taagttaggt104461aatatgtggt taaaggcatc tttgtgggtt tatttccctg aagaaacaga taaggttaaa104521tttctctgtt gggacttaat tttaactaac ttctctacag aaaaaccaaa ttatttattg104581caacttcctg tgaatctata attattttga aataaagact agaaagaagg agttttttaa104641aaagttctct gacttcagag gtaagggaga gggctcagcc tttatagtgt tggatgacat104701tatgtgataa atgcatataa agttcagctt cagtcagctt aagagaactc aggttaacct104761ttctctgtct ctagactggg attgagaatt ttttcgaatg tttgtttgtt tactcattca104821gcaaataact tattgagtgt tttgtatgtg ttagtgactt ttccaggcat ttgggatata104881gcagtgatca ggatagacaa aattttctgt cctcctgggt tttatgggat agcatggggt104941agggtgagag gcagagcata aacaagattg atcaagcata atttggaatc gtttcatgtt105001aaaacaaaat tgctaagctc tgtctccttt tctaggtcct tttcttcatt tctcattgaa105061cactttcccc acaattttac ttgagctgtc ttttcttttt aactgggaat ctccttcaaa105121tccatggaag atgaacctat attactcaaa cacagaaaga ctactataaa aatgaatcag105181catcctggtc ctatagattg tctgcataga taatcaaatt agaatgagaa agttgtttgg105241gctcataatg gtctaatgtc attggtcaga ttgcaaaacc acaatagctt caggcttcca105301tctccaggat gggctttgtg gttgtgaagg gcagatcttc ctttcctttg ggtcttgctt105361gaaggataca ttcattgagg gaacagctgc tataacaaat aggccccaca ttttaatgac105421tttgcacagt aggtatattt ctcactaata taatagtttg gcaggccgct gtcctcagca105481tggtgattca tgcaccagag ctccttctgt cctgtcactg tgccatcgtc tgttcagtgg105541gcacatagga aagagagtag aggagtgcta cagggtacta gtcctgcttg gaagtagggt105601aggccactgg tgctcacatt gcaatggcta caactcagtc acatggctgc atgtaacttc105661aagggaggct ggaaaatgtc atttctggtt gggcagccac ttagtgacac etetacacca105721ggcatagtag agcatgaatt attggaggag tctgctgtaa tcccacctat cagcactgcc105781accacttgct tctgtgactt ttttttctca gccccatgac tgaccaattt ccttgatctt105841tctttagaca aattgatatt cacttgggct atagtgtagt ctcaccagcc cattagctat105901ccttttcttt acagtagtca agtcttctca catctttgct aatgagatcg ttttgatcag105961caattgcctt gtggtagtct tatttccaaa atatttttgg cggaggaggc atcctcagat106021attcactgtg ggcttagtat gagcttttgc tactccaaat agtttgttta ctaaggcact106081aacaggacac tgttaaaaga tcattacccc tcataggtga aatggctgga ctctaaaaca106141gcatttatag gatgccttcg gggtaggttt atattttaaa cagaaatgtt ctatgattga106201aacaatacac acacccttct taagttctct ccaacaatgc ttccttgcaa ttcatattat106261ttagctttgt ttgattttta agatagtttg atagtagggg taaatgttac cttttgggat106321tttattaata gtattacttt ttaatgagtc ttttgaagtc tttcattata aacagcctgc106381tttccccaga gataataact tgccatgcca aatatttgca tttcacttga aacaaaacca106441aggggttgaa tgttttaaat ataagcaaat ctaattctcc tgtgtctaga gaaactgtat106501aatgaaccat atgtatgtgg cttaagcaga cagatgtgaa agatttctcc ctctctcctt106561ttatttaaaa aacaaaacac tggcagagct caaaattggc accaagtagc aaaYatctct106621gctcagccaa caaaaccact ggagctggag ttgatggggg gaggcctctt tagggtgcat106681gcaaaagtgg agcttttcct ggtcctggct tctacttgga aagctaaggc tccacagctt106741atttgccctc atttctcagt gccttcaggc ttcaaaatga gccattctca ggtgactgac106801ttttcagatc tttgagaaga ttgttgtggt tgattgttgg actggttcag aagacatggg106861tccacgcatc tgggtcctgc ggtgacatgg cctctagctg ctgacagagt ggtgctgagc106921taccaaacag actcataggc ttcctttcac aaagaacatc atagggcagt gtttccacat106981ctcttgcaat acaagacata cctctgaact tcctcaagct tgccaataag tagagctcac107041actacctttt agtatctgtg atgtacctgc cagggactat aggtgggata aagtatctga107101tatggctcct gtccttcaca ttgaaacctg ggtgaagatg tttaggaaca ctttaaatgt107161tcaaactgtg gtacttacca tgactatcat aaagattcat atatattttt ttgagacagg107221gtcttgctct Rtcacccagg ctggggttca gtggtacgat catggctcac tgcagccttg107281acctcctgcg ctcaggtgac tctcgcacct cagcctcctg agtagctgga actacaagtg107341catgccacca tgcttggcag tttttaaata ttttttgtag agatggggat ttgccatgtt107401gcccaggctg gtctcaaact cctagagtca agtggtctgc ctgccttggc ctcccaaagt107461gctgagatta tgggtatgag ccaacatgcc cagccaagat tcatattttt taagacgctc107521aaatcacttg gcttagtaaa tcaatttcaa gcatacagcg agaagcaagg ggaaagagat107581gggggggtag gttagcacac ttagtgcaaa ccagtagaag ggccactcta tgggttacac107641agaactcatg tgccctctcc ctttgtccta aggcctttct aggtttagga gtaggaactc107701tccttgccca cttaagtttt cttggaagag gggttgggtt ctcatgggaa tctaagaaat107761gagagccaag gaatggccca ctggtctgta ggggtgggag ttcttggctg ttcatgcttg107821ttctactgtt aacaggcctt gccctggttg tgttaagttt gctccactgt ccacttatct107881ttaggcttct gcagcctttc ttgccccccc ccccggcttt gtgtcctttc tgtacctacc107941tcccagcttg tgctgatgct tggttctggc gccctcataa ctttaagtcc cacctaccct108001ttctctatac tacagcccca ctgagcatct gtttagggtc tactttctct tctcattggc108061cttgatattt ctcatccaca ttcccaagaa tgagcatctg acctaagccc ttgtcagagc108121atctcaaagg ccagtatact gtgtgcccat gaacccttac tcccactcag gtgacgccag108181gtgtgaggtg ggtcatggtg cccacacaag gctgccaggc ccacctttgc agcagagcat108241gggaacgagc agtgccgtct gtcatggccg tgggtacggc aggtacattg gaggatttga108301gcaggggaag agtgttctgg gaagaagaca aacatcttac ccctgaggag tgagcatgtg108361cagtcggaaa gcctgctgag aggaggctgg ggtgcgaggc ttctgaaggc caggtggagg108421aggtggactt ttctgttagg cagcaggtca tctctgcagg cttttaagca gaggagttaa108481tcaccccctt ctgttcattc ctgatttctg ttgttagagt agacctgaga cctgcctgct108541aagaggccac taatgccacg ctggagtgag gtgatgtcaa gaggagctag aaagcagatg108601gttatggcat tgaggaaaga gcacacatgg gctgacaggg acagctaagc agcagcttca108661tgagcaggag ggaaagggcg tgttctaggg gagcagcagc ttccatgtca gactcaggaa108721tgcagcagac ccaacatcca gcttggagat ttcttctctt tttctttttt ttcaatggaa108781ctgtattttc gcaaaatgtt acagatcact tatagcaaac agaatggtga tggtcaagga108841aactgactct gggctccttt ttgaccgcag cagctatgtg gaagcagctg cagctgtgat108901aagggccgcc cagcatggat atttctgtgt cttcgttcac tgggaagtgg agggacagag108961cttccgaccc acatcccacc ttcttcacga atcatctttg ccaccaaatc gttggctcat109021aggtgtctgt tcttgtcttt cctgcccatg cctggctttt aggttccttt gctccRgctc109081tttctgtcag ggctgacctg gagggcagtg ctcctgtgag ttctccacat tatatcatgg109141ccggtaatct aatccaattc aagctgacat tctccaccct atgttctacc aatggaacct109201ggtcagacat ctgggctcat tcaggaagtg ctgctagggc atggctctgg aatttggtgg109261gagtattttg gaaggttgtg tggtctgttt ggttgtttag tgctcggact ggattttgat109321tttttttgct ttacttattt ttactgagac ggagtttcgc tcttgttgcc caggctggag109381tgcaatggcg cgatctcggc tcgctgcaac gtccacctcc caggttcaag tgattctcct109441gcctcagcct cccgagtagc tgggattaca ggcgcctgcc accacacctg gctaattttt109501tgtatttttt tagtagagaY ggggtttcac catgttggct aggttggtct tgaactcctg109561acctcaggtg atccacccac ctcagcctcc caaagttctg ggattacacg tgtgaaccac109621tgctctgggc ctgtttattt ttataataat aagaggtttt tgaaaaaaat tcagatagat109681gcaacccagt taaaaaaaat cccggtatac aaaatcctgc ctttgaataa gagattaaaa109741aaagatggtc attatttgct gcctgcaggg gttcactaag ggatatgggt aagttccctt109801agctcggaaa aaggataata attaataccc gcttacatct ccaaagtctt gaggtgctca109861aacaagtagg agtgtgtgaa atgctttgta catatcaaaa cactctgtga aatttaaggt109921attaactaat ggataatgac ttcttgaaca tttaagagtg gtttgattta cgtttagtag109981aaactcatct agtatgaaaa tgatacattt attacaaatg attaacaacc ctacagctct110041caacctcttt atgtaacctt ccttctgact aaaggaccca gatgactttg aggaatagcc110101tgtttgctgg gtgcattctc actcagcact tctggaagta tccattctac agttagtatt110161tcatctcatt gtcctggcct tccgaaagca cccttaaccc aaggtgtgag atatcaaagt110221aatacatatt ttacttcaga gatgacatct caaggtttta cagtagctca acattttcta110281gtctcctttc tctgaacatt gtcagagggg agacaggcgt tcctgggaag aattaattcc110341tggaatggtg tgtgtgtgtg ggtgtgggga atgggaggag atggccggcc ttcttttcta110401gggttagtgg gtggaattgc ccgcccaatg attaattccc tccttgacag tgtgttcaga110461agcaggcacc ttgatggctc tttagcagca ataccctcct cttctttcct agcctccttt110521tgtcataaag cctcatcttt tttatttatt tttatttatt tttttttttt tgagacagag110581tctcactctg ttgcccaggc tggagtgcag tggtgcgacc tctgctccca ggttcaactt110641ctgcctccca ggttcaacct ctgcctccca ggttcaagtg attctcttgc cttagcctcc110701cgagtagctg ggattacagg cacgcaccac catgcctagc taatttttgt atttttagta110761gagatggggt ttcatcatgt cggccaggct ggtcttgaac tcctgacctc gtgatccacc110821tgcctcggcc tcccaaagtg ctgggattac ttgcgtgagc cactgcaccc ggccaagcct110881catccttctg gtgttcacct cttggtcctg ttaaggtggc ccttggcaag tacctcttta110941gtcttagggt tcatgggagg gatagaacaa aggcccacgc ctccccctag catcgacatt111001tcccttcgtc tgccaatgcc taccatagct ttgttttctc ccgcactgct tccctggttt111061tctaggctct taaatcctca aatttgactt gcatgccctc tagtggcagg tctgatgcac111121cccttgttac cctgtttatt gtggtaccat aacagctgtt ctcaccaccc agctctgtta111181ctaaatattg tctgtaattc acaaatgccR aatgagaggt tcagtggtcc tgacttgcca111241ctcaagtagg agctggaaat gtgggatttg aattggagct tttgattcct ggatttatac111301agcatttcag tgtaacggca acagtaaaaa agatgtggtt atagttgttc tagtttcttt111361tttgtcattt aaaaagaatc cagattattt tactatccac tcagctaatc tgaaagaatg111421aaaatgtgag gaatagtgga aaataagaat gatcatcttt ttttaaaccc tttattacaa111481ttagaaattt ttttgtcact tactgagtat tcagtatttc atgcctcagt cctgttaact111541ttgaagaata caaaaaatac ataaagcatg atgtctgccc ctaagtttct gttctagtta111601gattctagtt atgaggcagt taaatttctt cacctgagga aaaaaattaa gcataacaca111661tgataacttt tattggtgtt taatttttct acttctcact aaagacaggt tctctgagca111721tttacttttt aatttattct tacagtcctt ttcagtggag agggaactac aggataacag111781cagttacccc gacgaaccct ggaggataac agaagaacag cgcgagtact atgtcaatca111841gttccgatcc cttcagccag acccaagctc tttcatttca ggtaagaatg tgggctccct111901aggcatcact ctcaccagaa catagccacc atcttgagaa atcattcacg caaaatgagt111961atggcagcct tctgtgattt cagtttgttt ttcatctaaa gtcgaatttc ttatcagatg112021taaaccatgc agaagaaaag ttgccaatga cgtttggaaa attatatttc catttcaaag112081ggcttaagtg ggaatttggg ggcaacttgg cccagaccct gcaccttatc agctgaggca112141gttcttgaaa tgctagcgtg tcgccccaag gggccacctc tcattgcttc ctcgttatat112201tagagctcgg gcaaggacag aatgtggcct cctaaggagt tggtgtcaca agtcttttta112261tgttatatgt gccttcaaac ttcaaactca ggctcgtttt ctttggctca aatgtttgtc112321ctccagtttt tctgagagaa attaaaatgg gggaggaaaa ttggccagtg ctttcccact112381tgtcagttct tctgggttgt cttcagatat atagattcct gttgagttct catgtaaaat112441tcaagctcag aggaactggt ttaatgccag tgtggctaca ttttaccagg ctgaaagact112501aacagtgaaa tagtagagtc tagaagaaaa attcataaag ctttcttttt agactttaaa112561atttgaagaa taacaatatg tccttaataa ttgtaacctg ccatattgaa gtgtgggcaa112621agattagttt tatcttgtga aatgaggcac ataattgaaa tatacttgta taagtaattg112681caaacatcat ttactgattg gtggtaatgg ctttattgac ggcagaatag tacagataca112741acaaattaat gttgacgtca tgattaagga aaataagcag aaacccatat tgcacgattt112801gaccacttgc taaccttacc ccgtgctcag gtgagtttct ttacgacatg tgcagatatt112861tagccaaata caaacaggta cttgttagaa atgtttttgt tttcttttaa cttttgtatc112921ttgcagaatt tcagatatag acacaactag tctggctgat acaacaaacc tctgtgtacc112981tatcacatag ccccaatgat tttctcttac ctacagttaa cacataaggt tgtttcagtc113041agtcaatgat ggcggtacca taacagtatc acggagctga aaaattccag ctccattaat113101cacctagtga tgctgtagcc atcatgatgt tgtagcacaa ggcagtgctc acataaatgt113161gatgatgctg gtaaaaacaa atctgtgcta ccagtcgtat aaaagcctag cacatatgat113221tatgtacagt atataatact taataataaa tggctattat tagcttatta tgtatttatt113281atgctatact ttttttcttt cttttctttt ttgagatgga gtcttgctct gtcgcccagg113341ctggagtgca atggcacgat cttggctcac tgcagcctcc gcctcccggg ttcaaatgat113401tcttctgcct cagcctcccg agtagctggg actacaggtg catgccccac accaggctaa113461tttttgtatt tttagtagag acggggtttc accatgttgg ccaggctgct ctcaaacccc113521tgacctcgtg atccacccac ctcagcctcc caaagtgctg ggattgcagg catgagccac113581cgcacctggc cttttattat gctatacttt tattgttatt ttagagtata ctcttactta113641tatataaaaa aaagttaact gtaaaacagc ctcaggcagg aacttcagga ggtattccag113701aagaaggcat tgttatccta ggaggcggca gctacctgca ggttattgtc ctcttctagt113761cacctccaca gtatagaggt ggaagacagt gttattgatg gtcctgacct tgtgaaggtc113821taggctgata agtgtttgtg tttcagtttt taaccaaaaa gtttatacgt gtttgtgttt113881cagtatttaa ccaaaaagtt taaaYggtaa aaaaattaaa aattttaaaa atagaataat113941agaataaaga tataaagaaa gaaaatattt ttgtacagct gtgcaatttt tttaaatttt114001taaatttttg tgggtacata ggtgtatata tttatggggt acatgagatg ttttgataca114061ggcacacaat atgaaataag cacatcatga agaatggggt atccatttcc tcaagcattt114121atctattaag ttgaaaacaa tttgactaca ctcttaagtt attttaaaat tgtacagtgt114181gtttttgttt taagccaagt gttattacaa tggagtcaag aaggttaaaa taaattttaa114241agtttatgaa gtaaaaactt acactctaag gttaatttat tattgatgaa aggatttttt114301tttttggtaa atttagtgta gcctgagtgt gcagtattta taaagtctac agtagtgtac114361agtaatgtcc taggccttca cagtcactca ccactcactc tctgacttac ctagagcaac114421ttccagtcct gcaagctcya ttcatggtaa gggacctaca aaggtgaacc atttttcatc114481ttttatacca tatttttact ctgccttttc tatgtttctt ctttctttct tcctttcttc114541cttYctttct ttctttcttt ctttctttct ttctttcttt ctttctttct ttcttttttt114601tttttttttg acagggtctt actctgtgcc taggctggag tgcagtgatg tgatctcggc114661tcactgcagc cttgacctcc tgggctcaag tgatcctccc acctcagcct ccctagtagc114721tgggactgca ggtgcacatc actatgcctt gcctggctaa tttttttgta gagaggaggt114781ctcactatgt tgcccaggct gttttctata tttagttgtg tttagataca caaatagtta114841ctattgtgtc ccaactgcct atagtattca gtacaggaac atgccRtaca ggtttgtagc114901ctaggagcaa taggttctac catctagcct aggtgtatag taggctatac catctaggtt114961tgtgtaagta caccctgtgc tgttcgcaca atgaggaaat cacctcatga tgcatttctc115021agaacgtatc cctttttcta gaaatggaat ctcataatat gtggtctttt gtgtctcttt115081catttagcat aatgttttag tttttttttt aattaaaaca tttgtgtggg tacataggtg115141tatatattta tggggtatgt gacatgtttt gatgcagaca tgcaatacat aataatcaca115201tcatggtaaa tggggtatcc atcccctcaa gcatttatcc tttgagttac aaacaatcca115261Rttacattct ttaagttatt taaaatatac aattaagtta ttgactatag tcaccctatt115321gtgctatcaa gtagtaggtc ttagtcattc tttctaacta gttttttgta cccattagca115381atccccgcct cgcccctact ctctccctgc cactacccct cacagcctct ggtaaccttc115441cttctactct ctgtgtccat gagttcaatc attttgattt ttagatccca caaatgagaa115501catgcgatgt ttgtctttct gtgactggct tatttcactt aacataatga tctccagttc115561catccatgtt gttgcaaatg attggatctc attatttttt tttaatggct gaatagtacc115621ccactgtgta tacataccac attttcttta ttcatctgtc aatggacaca taggttgcct115681ccaaatctca gctattgtaa acagtgctgc agcaaacata ggagtgcaca tatttctttg115741atacactgat ttcctttctt ttaggtatat acccagtagt gggattgctg ggtcatatgg115801tagctctaat tgtagttttc tgaggaactt ccaaactgtt ctccatagtg gttgtactaa115861ttcccaccaa cagttagcat aatgttttaa aggctcattc atgttgtatg taccagaatt115921tcattccttt ttatggctga ataattccat tatatcagta catcccattt tatttatcca115981ttcatcagtt gataggcata tggtgttctt ccacttttgg agtattaaga ataatgctga116041tttgaatatt tgtgtaccag tttttatgtg aacatatatt tttatttctt ttgaatacat116101acctaggagt ggaattgcta gatcaaactg tactgggagc tgaaagcatt caaattcaat116161ctcagtaagg tctaacattt caagaacagc tgggtcttgc agttatgtaa catgtaaagt116221ggaatgattg cctgatttca gttgatacag gtaaatcaac aatgaaggaa gaaagattcc116281tggataagtt tctcaaaatg ccactaggaa ttcttaaatc tcaccaaaag ctttcccaag116341atgtctgtct tatctctgtg tggtgtgatc aatggtatta attttcttct ttttggtcag116401ttttctatta cttttctatt tgaaatttct atttgaaatc tgccaaattt gttcaaagtt116461ttgaacttcg tcttctaaat ttttggaaaa attttgcaga ttttggtaaa aatgaagcag116521aaattaattt atgtgatttg catctttcat gagcattatg ttattgcttc ttttgcccta116581ttgtaattta catattttta aaaaagctat tcatgtgttg cttttggtcc catgattcat116641aattgctttg aggaataaat atgtttgtgg ttatacagaa gagattgctg aatgaaaaaa116701ttgtgattca tttgtatata aatgtaaaag ctactctagc tgagtaatta atggtttatt116761taaacaatgg cacttgacag gttctgtggc caagaacttc ttcaccaaat caaagctttc116821cattccagaa ctctcctata tatggtaagt acaattaatg ccatatgaca ttcataccat116881catccataac tcatccttta gttcctacct ttgcattttt taaaggctta tgttttgaat116941gatgttttta acagactaca ggaRtgaata caactcagat ttttgttgct gtgaaaatta117001aatgttatat gaaaagtaca atgaacaatg tctggcacgt attaagcact atgagtgtta117061actattattt taaagtaatt tcctaattat tatccccatc tcaatccctt ttcagtctag117121gcagttgtgg gcacatgaga cttaccatcc tgtagatttc attttcagac ttttctgcta117181ggtgtgatac aagtaaccag aggagcttga caaactattt tttatatttt ataaggctga117241gtgaacagaa ggttgaggaa tattttggag ttaggttaga tttgactatt ctagctgcct117301ctgagctttg caaatttggc tggtgtttat cagttgggcc ctaattcaga cttgcagctg117361tggtatccac tgcaccttta tgcactctgg ttccaaaagg gctatgctgg gatggtccag117421atggctctat ttgcattcat gaaaatgctt ggtgctgctg taaactctcg gctgtgtgtg117481ggctagttct tccccaaaac tgggcaggag atcgcactgc tataaatata taattaaaaa117541cattataatg tttatataaa gctttttgca tgaggattta cagaattatg ataggattta117601aaattgcttg tcatgttttg tgcatcttat tggaaaacta ctccaataat actcatgcat117661gtcaatgtat ctgggattct gagctggatt taggttcttc aacaatatat actctggcat117721aatttttttg tttttacaag ccttatatta gttttgtttt tgttttgttt tttgtttgtt117781tgtttgtttt tttggagatg agggctcact ctgtcaccca ggctggagtg cagtgacatg117841attatagctc attgcagcct ctgactccta agctcaagcg atcctcctac ctcagcctcc117901cagctagctt ttcatttttt tctagagaca gggtcttgct atgttgacca gtctggtctt117961gaactccttg cctcaggcaa tcctcctgcc ttggtttctg aaaactctgg gattacaggc118021atgagccacc atgcccggcc cttatattag attttctggc tagggaagaa taaagaaatt118081atcattttga aaagaatacc atttgttttg gcagaattcc cagtctaaac attgttggca118141tgagacctcc tatctttttg ttgagacacg cctatgaaag ctgaatggta ttctttcgat118201ctggcgtatg aagcagtttc ctgacccctc actctgggga acagctattt cagggcccca118261gtgcattcga aattggagga ggaggatgta aatggtctaa aggaaagtta tggagcttac118321cttagaaata tattttcttt caaatagtaa tacctttcta gaataatttt gtatttgtct118381tatctctatg cttgtcctaa cacaagtaca cttgtccctg tctacctaag agatggttaa118441gcttatgggg tcttagatat aatactttgg caaataagtt tcttttgtgc tgagtaaata118501ggtctgtgcc ttttagtggg aaataatgac tgaagttaag cctgtttgag actccccctt118561tcttcctttt tgtcttatct ggttccagtc agctttagtt actagtttta tcctaaaaca118621taattcattc cattttttat tgagcttact tgtgttttcc caggagggag ttctcaagca118681aagaacagtt tggtacaaaa atttacgtga cttgaaaagt tcacggtttt cctttaaagc118741tgaaaaacaa agagcttatt tctgtgctcc aggtaaactg cataatggct aggacaacta118801taaaatggct ggggccaccc ttcctctgac tgttttgaac accatgtcca gggcttttca118861ccccatgcag cctgcattta ctgaatgact gtggacttct gggtatcatt tttatttctg118921tgccatagtt tcttgtccta tgtcctcagt gttgtaagac acaccaacta gggaacctga118981agaatttaaa tttttattta taaaatgcta ctgatgctga tcccatcaat cggcattttt119041cttcctcgga gtggccccca gagatttcaa atggatttgg ctaggagggt ttgggtatct119101ggtagggcca tcaccacaga gtctgggagt tttattggca cgtttgggcc aagtaacttg119161ttctgtaagc cccatggtag gtactcatga catttgtttc atttagggag cttagtgatg119221ctgactgtga tggagccctg accctgcctg agttctgtgc tgcgtttcat ctcattgtgg119281ctcggaagaa cggctaccca ttgcctgagg gcctccctcc aactctgcag ccagaatacc119341tgcaggcagg taaggcctca ccatcaaYtg taaaccttaa gtacttttcc tccaggctgt119401gtaaaaaact tttgtttcct gactttttaa tgattgccat tctaactggt gtgagatgat119461atctcatagt ggttttgatt tgcatttctc tgatggccag tgatgatgag catttcttca119521tgtgtttttt ggctgcataa atgtcttctt ttgagaagtg tctgttcatg tccttcgccc119581actttttgat ggggttgttt gtttttttct tgtaaatttg tttgagttca ttgtagattc119641tggatattag ccctttgtca gatgagtagg ttgcgaaaat tttctcccat gttgtaggtt119701gcctgttcac tctgatggta gtttcttttg ctgtgcagaa gctctttagt ttaattagat119761cccatttgtc aattttggct tttgttgcca ttgcttttgg tgttttggac atgaagtcct119821tgcccacgcc tatgtcctga atggtaatgc ctaggttttc ttctagggtt tttatggttt119881taggtctaac gtttaaatct ttaatccatc ttgaattgat ttttgtataa ggtgtaagga119941agggatccag tttcagcttt ctacatatgg ctagccagtt ttcccagcac catttattaa120001atagggaatc ctttccccat tgcttgtttt tctcaggttt gtcaaactat gcagccataa120061aaaatgatga gttcatgtcc tttgtaggga catggatgaa attggaaacc atcattctca120121gtaaactatc gcaagaacaa aaaaccaaac accgcatatt ctcactcata ggtgggaatt120181gaacaatgag atcacttgga cacaggaagg ggaatatcac actctgggga ctgtggtggg120241gtcgggggag gggggaggga tagcattggg agatatacct aatgctagat gacacgttag120301tgggtgcagc gcaccagcat ggcacatgta tacatatgta actaacctgc acaatgtgca120361catgtaccct aaaacttaga gtataataaa aaaataaaaa ataaaaaaac aaacaaacaa120421aaaaaaaaaa caaaaaaact tttgtttctt aaaatgatgt ccagatactt tggggagtcc120481ttggtattta cagttattgt tctttaactg ctattagact ttcacatctc ataatgtgag120541gaatagggct gatctgggta agtgtgtatt acctgttttt ctcattctat accctctgga120601tggatttgaa ccaaagtcac ttaccctttc atgacagttg tgaaataggc aagtactgaa120661cctcccagct ggtataaaat gaagaggaag atcctgtaga gccaagtgtt aacttggcat120721ttttaggcac acccgagtgt ctccgggtgg ccctgggcaa tggagtggaa tggaactctt120781atacctaatt taaaagtcaa aaaattagcc gggcgcagtg gctcatgcct gtaatcccag120841cactttggga ggccgaggcg ggcggatcat gaggtcagga gatcgagacc atcctggcta120901acacggtgaa accccatctc tactaaaaaa tacaaagaaa aaaattagcc gggtgtggtg120961gcgggtgcct gtagtcccag ctactttgga ggctgaggca ggagaatggc atgaacccgg121021gaggcggagc ttgcagtgag ccgagatcgc gccactgcac tccagcctgg gagaaagagt121081gagactctgt ctaaaaaaaa aaagtcaaaa aattacatgc agtgaaatgt gcaaagtgtt121141gtcaccttaa atgtatgctt gatgaatttt tgtatatgta tacacccatg taactacaat121201cagatgaaga tatagaatat tctagcatat cagaaggtta gaatagaact gttatatttc121261tccttaataa aacagtctcg ctcaaattat tgttttggac cctttagatg ggaaagagtc121321atggtgagta taacatatgt attaattcct ttgggagagt agaaaaatta atagtgtgtc121381ttgccctttt caaaggcaaa tgggcactgg tctgatttcc attattcatg gaagtatagt121441gctcattgag aggccccaga aacagctggt gctttcaaca acctttctca ctttttaggg121501aagggctgtt aaagaaatga gagcttagtc atgggtgatt attagcaaga gagagcagtg121561tctaaagaaa ctttaattac ttattagtag agtatgaatt ttgtgattag atctgcaaaa121621ggccttagat tatcttgtct agccccttcc tcatgatgac accagattca aagtggcaaa121681tgtcttatct aagtttgtct cggggcctct gactctctta aataaaaagt ctgtcatttt121741ctgtgttaaa tagtgagagg agaggaactc aaccaaaggg cagaggccaa gtttgtggct121801gctgctaagt gacaggtaaa catgttgggg gaagagtgac ccattctggg atcttcttca121861aatgtcctca taaaatatgt gaaggggaga gttcagagag caaaacctac taatttctgg121921acttggattt tgctaacaat acctgctttc tgcctattgc aggctgtaac tctagcctgc121981tgcccttcct tagatgcttg gaagactctc atgggtccca ggaattagct tgcaattgag122041gagcatggga aactcctctt ttctctttgg tgtctttctg tctccatttc attctctccc122101tcgtcttgtc ttgtcttgtc tttttctttt cttttctttt ctttctgttt cttctgaatc122161agggcaaaga agcactggca ttctgggggc tatcttgacg cctacatggc tctcattgcc122221aaggcagtgc ggggtcatag aggagcccct tttcttggga ttggagacac atggggcttt122281tgtctctctc taaagggaat taatagctgg caaaRggcca agttctgttc tagggttata122341tttaatacct tcgccgtgtc ctctttcaag ccctggagag agtcctgggt aacctcatct122401cccataccca cactgagttt tctgtttggg ggtcatgact agtactgttt tattgttttg122461atgccatatg gccatggaac ctgtgggaca tgccatgact ccttatttga taagtcactt122521ttgtaggtgt ccaagtgttt cacagactat ttcaaaagtc actgacataa cggctgggtg122581tggtggctca cacctgtaat ctcagcactt tgggagtcca aggcaggtgg atcactggag122641gtcaggagtt tgagaccagc ctggccaaca tggtgaaacc ccatgcacta aaaatacaaa122701aattaaacag atgtggtggc gcacacctgt agtcctagct actagggagg ccaaggcagg122761agaatcactt gaacctggga ggcagaggtt gcagtgagcc aagattgtgc cactgcactc122821cagcctgggt gacagagcta gactccttct caaaaaaaaa aaaaaaaaaa gtcactgaca122881taatttttag acatctgaaa gccctgagct aaaagtatgt taacaggaaa gaagtaatct122941cttgattcct ttcctactgc caatggcttc ttttgacctg cttattcagt agaggggaat123001ctataaaaaa gagtgccaat gagccccatg tttcttcctg gacttttatg gagactgtca123061gcatgcattt atacaggaac aagggaagaa tccatcatat acacttcaat taaacaaaaa123121ttgcacatca tttctagggc ttgtgggatt aatcttaaat aataattata tagttctaac123181agaaaacatg aaagaaccca taaaactcta tcaaactcag tgacggtata ttttaaaatg123241ctttatcacc tatgtttatc attgtatgaa gcttaagaaa atggccagaa ccacctaggt123301ctcctctgtg ctttttttgc ttttctttgc aaaatatgag tgatcctatt catttttttg123361ttcattttaa tggcatctca tttagtgttt ttcttttttc tttctttctt tctttttttt123421tttttttgag acggagtctt gctctgttgc ccaggctgga gtgcagtggt gcaatctcgg123481ctcactgcaa gctccacctc ctgggttcat gccattctcc tgcctcagct tccaaagtag123541ctgggactac aggcacatgc caccatgccc ggctaatttt ttgtattttt agtagagatg123601gggcttcacc atgttagcca agatggtctt gatctcctga cctcatgatc cacccgcctc123661ggcctccgaa agtgctggac aacacgcccg gcctagtgtt tttttttttt tttttaaagc123721ttaactctta cctttttcca acttttttct aattgatcta tattcagtct ttaaaaaatc123781taatccagtt tttgataaga ttggtgaaga gtataaattt ggttgatatc ttcatgcatt123841cataactaga caagcttgtt ttctaaaata ataatttttt ttttgagatg gagtctcgct123901ctgtcaccca gacttgagtg cagtggcatg atcttggctc actgcaatct ctgcctcccg123961ggttcaagca attctcctgc ctcagcctct cgagtagctg gaactacagg tgcctgctac124021catgcctgac agatttttgt attttagtag agacggggtt tcaccatgtt gaccaggctg124081gtcctgaatt tctggcctca agtgatccac ccacctcggc ctcccaaagt gctgggatta124141caggcatgaa ccacccacca cacctggcct gttttctaaa ataatggaac aggaattgta124201tctgatgcca ttttacttgc aattatggaa atagtccctt ttccctaata gtaaaacttc124261aaggacagag ttgtattagt tggtggctta cccgctagcc ttttgatagt ttcatcaaaa124321gtctcctttt gatagtttca tcaagagtct ccttttgata gtttcatcaa gagtctcctt124381ttgatagttt catcaaaagt ctccttttga tagtttcagc tgggtggggt ggctcacgcc124441tgtaatccca gcactctggg aagctgaggc aggaggatca attgaggtca ggattttgag124501accagcctgg gcagcatggt gaaaccctgt ctccaccaaa aaatacaaaa attagccagg124561tgcggtggcg tgcacctgtg gtccccagct actcgggtgg ctgaggcagg agaattgctt124621gaacctggga ggcagaggtt gcagtgagcc gagattgcgt cactgcactc cagcctgggt124681gacagagtga gattccattt caaaaataaa ataaaataaa aataaaaaaa taaaaaaagt124741caccttttga taggtgattg ggtgggttct ggtagaaatt tttgccatag agtttgttaa124801aaaataccta tggagtactt actgcgtgag aggtaccaca ttgttcactg tggtcagaat124861gctgtatgac agagctccta ctcccatgaa cttgtgggca gttgatacgt gccatgaagg124921ggaagtacag gtgcagcaag agggtgcatg atcttaggga agatttcctt aaacgtttgg124981gctgagatgt gaaggctgag ctaaggacta cataggtgaa gaagagagaa gcttaggttc125041tgggaagggg gaacagcagg tatgctgggg aggctctgag agggggcttg gagcatagga125101ggagtgggaa tttcaggcca cagtggctgg cgtgcacaga accggggtag agaggctggg125161ttgaagctga ggaagaaggt ggggaacata tgacgggcaa taatgtcacc ttattttagg125221cagataagtg acacaatcag atttgctatg tgaaatgatc atgcaggtta ctgtccagag125281aacaggtttt gaggggcacg ggtagaaacc agaagactaa gttgtggtag ctggccagtc125341aggagcagcg gggtgcttgc aatggggagg tgacagtgat atggagagat gtggacacct125401tggagagaga tttagcaggt aattgacaaa attatagttt tggaaagtgt tagctttctt125461cacaggaaga tccgctcatg ataaagcatc agtcatataa taccagtcag agccttgatt125521gatagactag aaagagaaat cggtgaaatt cagaccagag taagcaaatg aacaaattaa125581ggtgcttgta ctcaactcca tgtggtgctg atactcagac cagtacttcc ttacttactt125641actgtatgta tgtatgcatg tatgtattta tttaaattag atagacacgt aggtaggtag125701gtaggtagat acaaagacag atgaaacagg gtctcactat gttgcccagg ctggtctcaa125761actcctgggc tcgagcaatc ctcctgcctt ggcctcccaa agtgctgtca ttacaggtgt125821gagccaccat gcctggccaa tggccagtac tttactgcag aaggatattg gcatttcagt125881ccctgatggg agtgggaggg tggggaagga ttaaaaacaa tgccaatcca cttttcacat125941ttgattcaaa tttttttaaa ccttgcatta caattttatt tcctaatagc ttgttgtgac126001aaatttgatt tgtagcttct gtttattatc tgccacagtc tgcttttcct ctttggaata126061ttgacaaatt ttctagctgt tgagatattt ctttcctaat tactttgtga caaagtagtg126121aagcagaatc cgcttctaag acatagcctg gtgtgaagta attagcttac caagtagcct126181gatctttcct cgtgtgaatt cactcaacta ccgaaataac agccagtgtc actattgcat126241attgaaagaa acccttaatg tgattctttt tttctttgtt gtaaatcagc aagaatttat126301gctcttattt ttatcattcc tgccaaccaa ggagataaat atttcgtaaa atcatgttgg126361gtgcacatca ttataagatc ttgcctattt ctaacaatag ttaaatgatg tatttcctta126421gttaatgtcc ctatacatta gaaactacac tagtaacata gttaagactt acatattgtg126481atagtgatgg ttattgtgga aaattactat gctataacat gaagtgttca gageacatac126541ccttcagcca gactgctgag agttcaagtc cctgctcttc catttatcag ctgtatgacc126601ttgagtaagt tgccctgttc tcagtttcat cacatgtgaa atggggataa tagaagtact126661gggttgtcca tataaagaac tgaaagtact acctcagcat gatgaatgcc atatgtggtc126721tatcagagct agctgatttt gttgttatat ttaacaagca cagaaaaRct gatgggaaac126781actgattact atggctatag gaaatattct gatatattct ttttgtttct tcttagcttt126841tcctaagccc aaatgggact gtcaattatt tgattcttat tctgagtcac tgccggcaaa126901tcaacaacct cgtgacttga atcggatgga ggtaaaagat cttcatatgc taataaataa126961ggatgtgtat ggagctaaat ccattggcct atgttcttta caaattctgg aaaatgaact127021ttttttttgt ttgaaaatga gcttttataa catttccttt tcatacttgg taattattaa127081tactttgatg ctcagccttt cagatctaga caccagatgt atctataatt tttttccctt127141ttctccctaa aactggatgt ggtaaagtaa gtaacataat ctctagacct tctagatgtg127201aaatgtagat atagccaaac taagacagaa aattctagcc aagttattta ctttaaccaa127261gctttaattc cctagtaaac aattttgatt Statttttat cagttgggca ttaataagct127321atactgaatc agaactgttt tgagaagagg tttgagaact ttatWtgagg gccccaaatc127381acattcggaa ctccctcagg gtttttagct tttgaaccaa atgaggactg atttggatga127441tccccagaag gaacttggag aattccccca gtgttgggcc atggatggaa gggcttagcc127501aagtgcttag gaagccttag ctatggatca ataaatagga gttgttatag ttattgttgt127561catcaccatt gtcattatca tcatcatcac cattatgatt actgtgatga caatttagga127621tacctgcttt tcccttcctt aaaagatctg gaaatgttta ataaatggag ctttattcac127681agactccata atagtcagag actattagag tcctctgacc ctagttgaaa ccactgcaag127741atgatagaag tcaaagtcag ggaacaggga catttcttta atttcttgtg actgtgcatg127801atgatttatt ctctcacatg aggccctcat gcctgtagca tgtgttatgg atggcaccgt127861ctgctcccct gtgagccagt caagcctgcc tttccagtgc ccctctgact aacatctctc127921tgccattttg cttctctttc tttgagttgt gtcctccatg gtactttcct cagatttcta127981ctcatgggaa aaagacataa agaaaagtca cttcaggagg aatgacagat gtgcatttga128041agaattttgg ttaaaatttt ttttttctgc agaatgctta tgacaaagag agagataaaa128101tagtttgcca cttttcatgg ccctcagttt actttggggc ttctagcagt gtttaaagag128161ctttgtttat caagccagga tggatacatc tatggctctc tgactatgtg tgtgtgtgtg128221tgtgatgggc cccctgcttg cataaaatga tgaagttatt agttggtctt ctttaaaacg128281gccataaaat ctgttacaca tgcacacgtc tgttttactg ttacacacac acacacacac128341acacacacac agtctctctc gcttgctctt tctctctctc taatgtcctg gatggcgtta128401catatattca cctttgtgga ttttatatgc agctagtttt atatactttt gcatacttta128461agtggcctcc ttggggaggg gagattaaaa actacctagt cttttttttt tttttgccct128521atccaaagaa gaaagcaaaa caacaacaaa aaatcccctc ttttccaacc aaattgttat128581ttttgaggat gctatggttt tagacaatga atgcagtaaa cagaattcaa ggacaaagta128641atagggttta ttgatggttt gttcatttaa gaagtacggg tcagattgca ataatgattc128701aatatttcat gttcttctaa actattactg attgttcttt ctactagtct tttaatctat128761catgaatgca tggagtttaa ttagattttg gaaccagccc tgtgatcgat gactccagta128821taggtggttg tgttctgaac aaagctgcag ctgatttagg tctcaSgtca tgagaaattg128881gcttagtgat acatcttttt ctaaagacag cagagctgtt agccatgtac ttatttccat128941taagaattac agatgataat ttcaagtata tttcttttta tgagtgtctc ttacattact129001ggatttcttt ctttttctaa acagctttat tgatagagac tttacatacc atacccattt129061aaagtataca attcagtaat ttttattata ttaacagttg ttccaccatc tctacagttt129121aattttagaa gaacatcatc agcccaaaga gaaacccaat actcatgagc agtcacttcc129181tattccactg tctctgccag cctctgataa tcactagtct attttctgtc tctagatttg129241cctattctgg acatttcata taaatggaag catatagtat gtagtgtttt gtgaccgtat129301tctttcactt agtataatgt tttcaaggtt cactcagatc ataacatgta tcagtgcttc129361attccttttt aatgctgaat aatattgcat tgtatggata tgctacattt tgtttgtcca129421ttcatcagtt gatggacatt tgcattgctt ccacttttgg ctattgtaaa taatgctgct129481atgaacatag atgtacaagt ttttgtgtgg gtgtgtgttt tcggttctct cgagtgtata129541cctaggagtg gagttgttgg attgtatcat aacttaactt tctgaggaac caccaaactg129601ctttccaaag caactatact attttacatt cttccactag cagtgtatga gggttccaat129661tttcccacat ccttgttaac acttattgtc tgcctttttg attttagaca ccctagtgga129721atgaagtggt atctcattgt ggttttgatt tgagtttcct aatgattaat gacgttgagc129781atctttttgt gtgcttatta gccatttgta tatcttcttt cgccaagtgt ctcgtccttt129841gtccatttta aaattgcatt acttgtctat tgaattgtaa gtgttcttgt gcttttggca129901tcatatctaa gaaaccattg cctaatatta aatcacgaag atgtacttcc atgttttctt129961ctaagagttt tgtttgtttg tttttttgtt ttgttttgtt ttttgagacg gaatctcgct130021ctgtcaccag gccagagtgc agtggcgtga tctcagctca ttgcaacctc tgcctcccgg130081gttcaagcaa ttctcctgcc tcagcctcct gagtagctgg gactacaggt gcgtgccacc130141aagcccagat aatttttcta tttttagtgg agacggggtt tcaccatgtt ggccaggatg130201gtcttgatct cttgacattg tgatccaccc acctcggcct cccatagtgc tgggattaca130261ggcgtgaacc actgagcctg gcctcttctg agagttttat agttttagac ctttacattt130321aggtgtgtga ttttgagtta atttttgtgt atgttctgag gaagaggtcc aactttattt130381ttttgtatgt ggatttctag tcgtcccagc accatttatt gaaaagatgt ttttttcccc130441attgaattgt cttgcaccct tgttggaaat caactgacca tagatgtaag ggtttatttc130501tgggttttca gttctattct attgatctat atgtttatga ttatgccagt accacagtgt130561cttgataatt ttagatttgt agtaagtttt gaaatcaaga agtgtgaatc ctttaacttt130621tttcttcttt ttcaagaata ttttgctatt ctgacttcct tgcatttcca tatgaaattt130681aggacaagct tgccaaattc tgacaaagat tgcattgaat ttgtagatca gtttggagag130741tatggccacc ttaccaatat taagtcttct aactcatgaa catgagatgt ctttccattt130801atttagatat ttaatttctt tcaacaattt tttgagattt ctttttatct acattgccta130861cttctaaatt atgttattct ccaaaggggc ctgatttata atgcaggtgg gaaaagacaa130921tgaaagaaat gcactgttat gatacattgt ctgttctatt ggagatagta gactttgaat130981agaaaattat atctggatac ttcgttttaa tgagtaaatc ccagcactat gtgaggccga131041ggtgggcaga tcacgaggtc aggagattga gaccatcctg gctaacacgg tgaaaccctg131101tctttgttaa aaatacaaaa aattagccgg gcgtggtggc atatgcctgt agtcccagct131161actcaggagg ctgaggcagg agaatcactt gaacctggga ggtggaggtt gcagtgagcc131221aagattgtgc cactgcactc cagcctgggc aacagagcag gactccatct caaaaaaaag131281aattacctga gtgaagactg ttaaatctta gaccagtttt ctactgagta atacttcagt131341ctctttcttc agaacagttt aaaaacaaaa gtattgctta tctctctggg gttgtttagg131401ttctgagagg taaaccaatt aactttttgg aacaccagat ggctccaggc atttactctc131461cttccttcaa gataaatgtt acaatttatc tttatttaat ttgtgactat gttttaaata131521taaacaatat caaaagacat atgttgacaa gtcttcctcc catctaccaa ctcacctttc131581aattcttaca tatctttcca gagtttgtgt atataaataa agcaaatgtg aatatatatt131641attcttgaac ttaatcttat aggcattaga gtgcatattc ttttccagct ttgaggtata131701attgacaaaa aatatatatt tatgatgttt tcatctatgt atatattgtg aaatgattac131761tgcaatcaag ctagttaaca tacctgtcac ctctcacata gttacctttt tttggtgtgg131821tgagaacatt taacatcaac tcccttagca gttttcaagt ataccataca ttgttattaa131881ctatagttag aatgtgtgtt aaattgagaa tttacaagaa gtaaatttac agtaagtaga131941ttttaaacgg tttaaaacta attcatactt gatagatttg ggtattcata ttagtgtgag132001gacttccccc ctgtttgcat taggagaaaa ttttaatctg ctcatcagta aaattctttt132061atcccacaac aagaaggctg gtttaatcag gtttagacac catctgcatg acagatcttg132121gcaaacaccc acattgtggg gccacttgtc agaacttctt tgtgtttgtg aaaaaataca132181taaagaacag gtttgttctt ggccaaatgg accagagttg ctatgcaatt ctgtggctga132241atcccagtct gaattcagat ttaatacatg tgccttaatc tgaatcattt ttttcctctg132301tgtatgtaaa atttcaatac aaattaacat atacctcact aggaacctaa aaaaaataag132361gtagtattaa atgcagggtt tactgcagat ggaaatggtg gcaagaaata atagctttta132421agacattttc ggctgggcgc ggtggctcac gcctataatc ttagcacttt gggaggctga132481ggcgggtgga tcacgaggtc aggagatgga gaccatcctg gctaacacag tgaaagccct132541tctctactaa aaaatacaaa aaattagcca ggcgtggtgg cgggtgcctg tagtcccagc132601tactcgggag gctgaggcag gagaatggtg tgaacccggg aggtggagct tgcagtgagc132661cgagatcgcg ccactgcact ccagcctggg cgacagtgcg agactctgtc tcaaaaaaaa132721aaaatttttt ttcatcatat gtgcgcatca cttaatttct gttccaacca gtttaattca132781tttacttttc tttttcaaag aagacatctg ttaaagacat ggctgacctt cctgtcccta132841accaggatgt aactagtgat gacaaacaag gtaggagaag aatgagtttt cttctttaac132901cagcgttcta cctgttacct tatctactcc tcttggaatg gtggcttcta gagcatatga132961gggtggttct cagcatgtaa acacatttgg atagactaag tagttcaaat cattgtcata133021aaacaatagt tcacaaagtg gtagcaagag cttgagtgtc cagagtcttc caactcattg133081gccccttagt cattgctgac cctcagccta ctcttaacac cttacactgg aatctgatcg133141tagcaattag gaaatgacag gaagctgaac Raattgagaa catgttgtgt ttgcggatgt133201gatcccagac atcagtgtaa atgcacattt gcatacagct ctaatccatg aggaagttct133261cttctggaca cagcttctgt ccttagtgtt tatctgtttc ttggctttgt gagtgtggga133321gtgtgacatg gagcccagct acaattttcc tatcttactg ggcagctgag aagcagtgca133381gcacagcatt ccgtgttaga gctactccca gggaagagtt ggcttgggaa tatctagaag133441tgtgggaggg accctctccc ccacctccta tactttgtat agtgctttat aatctaccag133501gtccttgctc atatactgtc ttattcagtc ctcacaaaac tgaatggagg ctggtaagtt133561ttatcacctg aaatttagag atgagtaaac tgaaattcag acactatgaa tcatttgcct133621agttattaag aaaaagattt gggacaagaa ccaagatttt ttctaagcct caagccagta133681ttctgcctgt tatttgatat tgatttacta cccatgaatg agaggcttat ttgaagggac133741taaaaatcag tgaagctatt tcaggtttta ttgttaaggc ctgccattga ttaggcattg133801tatgggggct taacatgaaa aataagtttt ccattaagtg acagtaattg catgttctct133861atttaaatac ttgcttggga gagacaggta gaactggaca ttgtctgcag tagcgtgtac133921tctgaccatt ctcctaactg cacagtggaa taactgggca aactgtaaaa atattgaagc133981ctggatttca ctcccaggga ttctcatgtc atcagtctgg gtgtcaggaa tgaaagaggg134041agaaactccc aatacttcct ggtaatactt ctcctatctg cttgaggtgc tgtctctgga134101aagttgaaaa tacatctttg ctctttgtag aaggagaacc tatttccata ctccaggagg134161tctttattgt tttagtaaat gctatagcaa cttttagagg tttgtgagac ctgtttttgt134221aagtgatgac tagcgattct ttgtttgaat ccaatttgcc tcagaaataa cattttatag134281ttttaatttt ttttcctctt ttctctttct ttttgcttaa aaaacaaaac acaactagct134341ttgaaaagta ctatcaatga agccttacca aaggacgtgt ctgaggatcc aggtaggaga134401aaaccatttt tgtatacttt atatataatt attttaaaat aaaaaaacag caagtcagtg134461agtagaagat tgatacctct gtgatagtgt gtctgtttct ctctatttat ttgtcttgta134521ttttattaca aggaaacctt ttggcttttt accctttctc tcttttcagg tatgagaacg134581gggtgatatc ttttcaagtt tttaagtttc actgaaagga tcacgaagga tcactccgtg134641tgaatatagg gtcttgttac taagtgcaaa attgagaagg cttgaggttg agatgatttc134701aacaagccat tttgggtttt gtgtagagta aggaacttat agggtagatc aattggaggc134761ccagagttgg tattttccat ttccaaagca gaaatctcaa atatttttat tatgcttcac134821caccaggaag gcaactatat gtgcttatat atggagtgga ttccaaggag aaaaataaaa134881gttgtccagt tttttggaga gtcagacctg tgtctgtgga gtcatggaga atggctgccc134941ctgtgtccgc agctctctgt tgatggagcc atcattgaac caagttgaat ctagttaaaa135001acaaagtggc tacagaagac attactattt ggagctctag ttataaggac atgaaaaaag135061ttgttgcttt ctcttttgct gttttatttc ttgtatattt ttaataagtc aatcatgttt135121gttgtgataa gctagctatc atcctaggct tatttgttga acattggaac agaggaatca135181atataatgtt gaggcttatg ctgtggtaca atattttcca aatgtcaagt tcattcagag135241tttcaaagag agatgggtgg agggagagga aggagttggg ggagtgggag aaaaatatgg135301tagggggaag gagagagaga gagaacatga gaatgagaat aaatatgaat gaatgtggat135361gatgaatttc aattatggaa gcctgagaag gtgtattttt aggcagctcc ccagttggtt135421ctggtacata gcctattttg ggaaccacca gctctggaag gtgatgctac aaaatagaaa135481attaaggtta tagatcaacc atgactccaa aagtgatggg aattcccaag cttaattttg135541actcttgaaa cagatttgaa ttcattcctt attgggtgaa atgctctgtt aatgtaggcc135601aacgcctggg gtcagttctt tctagctctt atttctggag ctaaagtgga tgcattccag135661tttttgtaaa cttcatgttg tagtattagt aaaagctgtt taaatttgga gtgttaaatt135721ggtgcttctc caacagcctt tgaggatcat aacttttcct actttttttt ttttttttaa135781gggattttag cacaagatca agtgtcctct ggggcatgag ctttagatct catttttaac135841attccttcat tgactaggac atggcatcac caaataaaac ttgtcaggtg tgccacagca135901tgagaagtat gtcggaattg atttctcgtt gtgcctggca gcatgctatg tgttttctga135961tttgttcttc ccactactgc ccctctctgg gtatgaggga gttcctcatc ttaaacatag136021ttaatccctc cttttctggg aaatgtttgg attcccagtg ggagacacaa gcgtttcaag136081gtggtgattt agaattctgg atggctgtcc ttgccatgtg gacctcagtt cagttgctgc136141tttttggagg gttaatctct aggcctgctc ctctttccat tcccatttcc tggaaagatg136201acatgccttg agtgttctag gaaggtggca gtccttttcc ctctttccag ctacacgaga136261agctatgatc cacttgttgt aattgctgag agtgtctatg ttagtggagc cttaagagat136321tttttaaacc aatatcccat tccaacctaa ttcagcagag caccatgcac aatgRtggtc136381tttttcttga tgtttctggc ccatgttgag gtgttttcag ctggttcacc ctcacagtgc136441tgtctttaga ctgttgttgg tgattctagc agctgctcca ttgccaacta tcaccccatg136501tttttattgg agtagagcag ttctgtcctt Scattacctg ctcctacctg cacctccttt136561gctgtcctag aagaaactaa atttagaaac cagatttcca ggcttaaaaa attaggagcc136621tttatttatg cactctacaa tagtaatatt ttgcaattct aaagtacgtt cagtttttaa136681attactttca ccttatctgt ccctgtgaga aagaaactgc atttttattg cctttgtttt136741ctaggtgagg agactgaggt tcagggaagt aacagctagt tagtgtcatg gccaggagga136801gggccttagt ctcaggactc ctggcctggt ctctagtcct gtctctccac tactcatcca136861caccctcacc ccatctgtct gtctgtctat ctctYaccag cagggctcac ccctccttcc136921acacccatcc ctaccagcaa aaagacaaag ataatggcac ttgacaaagg aattaacagt136981caccatggtc tgatttgggg aacagtattt tatcatggcg gggacctctc ctacccatgt137041atttcttgaa agcagtgggg cctgagggag aagagggcca cggagccagc tgcaggcagc137101ctgcgtagag ggcttcccca gctctgtggt gagtctggtc cactctgtcc acccagcccc137161aaggcagcct cctgagctgg gagagaggct tcctgaggtc gctttctttg ctgcctgggg137221tttgtaggag ggagggaggt ctttttctgt ctatatccag gtgtgggtgc acgataatgg137281gcaaaagggg actcaaactc cttgggcttt ccaggctttc ctgcactctg gcagcgtttc137341agctgtttga agttggaaga tctctgtgtt ctcactacaa caagccaaaa tgcttgggtt137401gggagaagtg tatctcttgt gcagctgacc agtagtttgc tttgtgcctt gttagcattt137461atgaggattt tcacctcccc tctctgtcct gttatctaac ccaaggattt taatgatcat137521cataaattca ggctgcttca taccaaggag ggtttttatc tttggagcaa tgtaaaagag137581gatgagataa tgcagtgaga cattttcttt ttggaagaat tcatgcagac aaagaaatta137641cattcttctt gttctgtgtt actagcacct tttctatgat tttttatgac attttatgat137701gaaaaatttg caatgtttgt aacacctgag catagctact ggttcctaaa gttgtcttcc137761atagaggatt gaaagcataa aaagaggatc agtgtttttg taaatgtttc taattactcc137821tcacaacttg tgatggggaa gaaggctgta tgggcgttgg tagtgagcaa gttagtttat137881gatttaaatt tgatgaggaa aagtcatatg gctgggaaaa gattcagcga tattgttcat137941aattctctgt atgatttggt cacagacatg tgggacacaa ggatattttc ttacaggaga138001agctgtatag ctcatttcca cttacttgtg agagctttgt ttttttgtgg ccatgctgct138061ctggtagtgt attttttttt ttttaaaacg tgtagttaga tagacttctt cctatccttc138121agggtagact agttcaaaat ctattactag actatcatga tagaacaaga aaccctgaag138181tgtgaatcca tcattatata aaaagacaac cccctttcaa gggggttgtc tttttatact138241ggaatgtctt cattaattaa cttggatttg agaatggcct ctcaaatagt gcttgaccaa138301aggaaaaata tgctggtttt tctttccttt taatcgtttc tttagatttt gaaataaaac138361aaggaaatca tgaaaatgtg ggagccgtcc agttctccac atgggcactt ttacctcaca138421tttacctKtt gatgtgttct gtactagccc aggtgctgca tataaacttg tttaactgca138481gaaatgaaac ccaagcaata tctttgtttc agcaactccc aaggattcca acagtctcaa138541agcaagacca agatccaggt agtgttcgtt taatttctgc tttaatgccc tttgtgccgt138601gcccctgcct agaaataaga accacaaacc aaaacccaac ccatgatttc ctcttccttt138661agatgtaaag cttaggatat gagccatttc gctcgtgcat ggcctgatgg gaacatgcag138721taatgctggg agccttggtt tctgtggtca ttagtgctag aaggtgacca tgaatgccct138781acggcccctc aacgtgaagt ggattgtgtg agacaaccca gttgttgcaa gatgataact138841tataggggga agtagctttg gaaagaggaa aatatgggta aggtaacaga cacgaaaatt138901tgctatcatg acaaggtgaa caccaaaaag taaaaagtct gggtaataaa atctacagta138961gggaaagaaa atgcttctga taacttcagt gaagcttgca gtttcaggta agcgatgttt139021atggtctgtt tgagcctatc ttagtgcccc tStaagcttc acatttttta agaggctagt139081acaagagaga aagccagtaa ggcagagctg tggttgatgt gagactcaga agagaggtgc139141agaggggaaa tacagagaca ctaatagaat taatccatta aggaagggaa atggtattgg139201tactgtattt cactggtaga actgcagtgt aagataggat aacgaggata ccatRtgtgt139261gttttctgtt ttgctttggc tgttagtgac atttttccct cctttgccta ctagacacaa139321tcctgaaaaa gtaaatttaa gtctgatcat attttatatg cttgaaatac tgatatttaa139381ggaacctcat ctctttgtaa gtcaaagagt tatcctttag tttatatgtg cattttccaa139441attggggttt tgtttccttg tttatgtaag atacagatat ctatattttt atatttatgt139501gtatttaaaa acaatagagt aattcattta ggtaggaaga aaaggtgtag gcattctttc139561tgtggtcaac acatttactc atcctttgta aaaggatgaa tggatctttt acaaactagt139621ctgtactagt tgtgggatat gctttcatag taggaattgc ctttgaattg cagaaatctg139681ttatgtgtac tatgttaacc ataaatcgaa cctcaaagtt ttagcttctg ctgacttcct139741ctctccctga aagccagtga aattataaaa tctcatagcc acacgttctg ttattaatgt139801gtctggctaa attttacata ttatcctttc agagattgcg cttttggtct aatttggagg139861attttatttt cattacttct cactgggaag aagagaattg aattgttttt atcctccttt139921ttgaaataat aaagttttgt agaaatataa gggaaagagg gaaaatcacc attttaacac139981cactgcattc atcattttgt taactccctg cacatttatg ttttaattgg ttgcagcagt140041catggttata caatctcact tattttatat tcagtaactt tccatatgaa gatataaaca140101tacaggttat atatgaaaat aaaatgaaaa tgaggaatgt gcgctgtggg attgtgtgct140161taacagtgtg cacgcataca cattatttac acacacatta tttctttttt gagtgaaaga140221tgtcactaaa gctatatgtg gcatagaaag ctcttttcat ttctagctag ggtctctaat140281tgtctttttt aagtgaagtg ttcccattta agaaccttgt ccttccgtat cagttccctt140341tgattcttgt tgaaagagcc agagcattct tccctgctag tttctgcctg ttgctgcctt140401gcctgtcttg ccagttatgt tacagggacc accccagaac acgtcacatg ccattatggt140461ggctttatta aagttacaac cttgtaagac agacagaacc ctcaactacc acaaagaatc140521cttgagaaat ttattcagga aaataaaaac tcacaatgtc aaaaacagct ctgtcaagaa140581ctgaaggggt ttaatagtaa ctgatatatt Staacttcat aaatgccact ctaatagcta140641agtacgaaga aggttttcag aggttaattt gtctctctag gatgtacctc tgactcagag140701gtcgttttct gctttgacat gaagctagtt gagaagctaa ataaaacaga tgtatacatc140761tacatcttag ggccttttcg tgatcatcgt attgggaacc cgtccttaaa cctattccta140821agcagtgtat ggagttgaat tttatttgga aagtaagtag gtctccaata aatattctag140881ttgctatcac caaagtattc ccccgccaac tcataggcag tatgagaaca ttttaaaaat140941gggatataga aacagagatt ggagtttgaa tcgaactgtg taactttcta tctgaccttt141001tgcaagttat ttagttcctt tttaaaaaga tgatattacc agagttacaa tctcagaagt141061tgcctagtta attatgcctt gctatagatt gaggaagaag gttgaaaatg agacttttgc141121tttctcaggg aaagacaagc tagacccctc aatctgattg atgtgtgttt tcctttccca141181gcttcgtttg tggcatattc ccctgaacat tgaattctta ccctcagctt atctgtccca141241attccacatg agcctgtcat cacatttgca agtgaacagg agaaaagttc tctgatcagt141301ttataccatg aaataccatg aagcccttct gttagaacat ggaaatgcct aagctgaggg141361gctccatatt attcttatta acaattccaa tagaaatgga tttgaattta cacaaagttt141421tttttggggg gttgattcag atctaaggta taataagaac aaagaggttg aaaagatccc141481aattcatagt tggatttgga caattattgt acatagtggc attcaatggg taatagaatt141541tatggctgct ttggtagctc atcggttctt tgtagtcact ttgacttcgt gggtattttc141601catttcaccc caacttcaga gagtaaagct ttgctatttc gcttctgacc ttcccttatt141661ttgctacaga tcttactcta gcacctccat agaagaggcc atgaaaaggg gcgaggaccc141721tcccaccccg ccacctcggc cacagaaaac ccattccaga gcctcctcct tggatctgaa141781taaagtcttc cagcccagtg tgccaggtga agtgcccctt gccccctgag cccttccatt141841tcctgttgga gccagcggat gtgtgtctgc cacagtggcc tctgtgagtc cggagaaaga141901gcagacctgg cagtttcgca ttccgtgaag gtcttcacgt ggctatggta ttcttgggag141961agaaagcaat tctagacaca tatgtagcac atgtgtctgg agccttgccc acacttgtta142021tggatcaaga attaggaaat tctctaccaa gaggataaaa atgcttcact cccatgcagc142081ctgctgagtg tgtggctggg gtcttactct ctgcatcatt ggtaatttgt gaacaagata142141gatSgctgtc tgtcatttcc tcagatgcct gtgtggtctt ctatgaaaga tcttggagat142201taaggcctag ccagaacctt ctcctgcatt ttacacttat ggagaagatc ctgtgtcctg142261taggatttaa tgctgagggg actctgaatc tgactgatgg ttgggtggtt ggtggagggt142321cctagggcaa gggctccttc aggtgggctg ctctgggctt ctggggatgg gatgttcttg142381agcccgcagt tgtgtagtgg ggctggaggt gggggaggcc cagaggggcc ctgtctgctc142441ccttggcttc ctgtgaagta gaagataccc tcctaaggaa cctaaggaat agaagatggg142501aggcatgcct ttgctttagt gaaggtgata ggatacggtg ctgtcccaag tgttattttg142561ggagcagcaa gggcctttcc actgactcat gcaggcaagc tataaataca atgatttcct142621ggaacttttt ctccatctat gtgcatttta aatcaggcca gcctcagatg ctagtgattt142681gcagattatt atcttttctg ctaaagcaag aaaagacttt ccttggctca cacattagca142741tgacctctgt gtctgggcag agctcagcat gttttatttg ttctaataaa gtattctcct142801cacagtaaag ttatagattg ttttttaata tactaaaatc tggaactctc tatcaaattt142861aattctgtgc catgagttca tcagacaaaa atttgccttg aatttcctct catactttga142921tctgaattct ggcataaatt tcatagtgac tggtatgtag ggtgggagta accacaggtg142981atgatgctat aaaaatttca catggctggc ggattatgac ctggaagatt ttcctatgga143041tgcgagaaaa ttaaaacaat ctgggaagct atctcagtac ctttaccttg caagaaggga143101ggctggaaat gctcacaggt ctctctcagg ttgtgtgacc cagtaaggag atgccagctg143161tactggagaa ttcacatgct gtggttttta cgacactgca tcacatagtg gaagagacaa143221aactgggttt ggagctcagc tcagctgctt attcactgtt tgattttcag caagtagctt143281aatgtttatg catctcagat tcctaatctg tacagtagag ccaatactcc atggagttgt143341aaggttttgg tgagataata aacataatcc aaatatgggt tgaatattcc ttatccaaaa143401tacttggggt cacaagtgtt tcaggctttg gatatttttg gattttggaa tatttgcatt143461atatgcttac caattgagca tataataaaa aaatctgaaa tgcttcaatg accatttcct143521ttgagtgtca tgttggcact aaaaagttta gggttttata gcattttgga ttttggattt143581ttgggattag ggatactcaa cctgtagtac tgtatttggc ccacaggcct tagtaagcat143641gtaaggccca tattcccgtg gcctgcaagt atcaagaatt taatgcctga aatggcaagc143701ctccaccgag gaggataaca ccccgacttc cttgacccct agcagacaat tctgtgattc143761ctcctaccca gtttctcaga aggctcgcaa aaggactgaa ccccagttgc ctgccgtagt143821acaccattca ttgatgtact ttttcttgtt ttgcttccct cctgccctgt ctcactttcc143881cctcactcga ataatatacc tgttctcaag ttcatatgtc aggggcttct ttagagaaaa143941cctgaattaa gacagctttc tttgatatct ctgtgtcaat gtgacttcaa gatgaaaagt144001attttggttt gataatgccc tgacgatctc ccttgtggta gggcctgata aatctaagat144061ttagacaaat aatagctagg tgggtgatgt ctctgttcat agcctaattc cattgtcttt144121tcagtacaaa tttatacaga tgaaatttta atgagcagtt taaatttatc acatgaagaa144181tagaacattt ctgcagtatg aaaagcaggt ggtataattg catctacttt atttattttc144241atgttggtta attctcacag aaaagtataa gggtctgtct aaattatata aaattttagt144301gtattgcata ttgctagaaa ttatcgaaat gctgacaaac tagaaatgca tgaaatagag144361cttaaaccta ccagaagagt ctactcactg agaatcatgg tgtacattta ggcatccctt144421tgctgcactt atctttatgt atcactttag ataaatggga tcacactatg cttgctgttc144481tgtatcctga tctttccaat taaccatctt tctatttcat atagaggatt gcaccgttcc144541ctttaaggtg tgcatgatat ggatatacca tggttgactg tgctttgatg gagagatctc144601tcccttcttc tctctgcctt ttctcttccc catactttcc tcttaagctc acccaccacc144661ccacgacccc agcccccgtt cctttctcac ccttgcaaac aaacctctct ctctctccac144721cccgaccccY gtgtcctctt catacccctc tgtttctctt tttgtccttc tcccccatca144781ctccttcaca ggtatgagcc tcccctcact ctccccttgc caagctcccc cacaccacca144841ccttgttctt ataatataaa cgaggcagag gttattaata atttcatgtc cgtatctttg144901tttcatcatc taattatttt cttaggttaa atcctgctgg ttggagttgc agggtcaaag144961gctgtagatg tttaaagtgt agctatatgt tgccaaaggg tgaggtggaa cacctgtcaa145021cttgctccat catcaacact gtctgaaagt gtttcctcac aatcttgcca acactgcttc145081atgttcatct ttttaatcat tacctgtgta ataggcagtt tcatgtccct gattcctagt145141gaggctgaga atctttttgt attgttattg gccataaata ttccttctct tatgatttgc145201ctgctcagaa ggcttggcca gcttctgttg ctacatctct ctttgcctaa ccacggggac145261cacttctttc ttcttcattt tgcctacgtt ctggacctcc ctcctgtcag gcacttggga145321atgtgagggt ggccatgggg aagcctacct cacctgctgc tgtgctgttc cttgtccctc145381cagccccgcc ccccatatcc cctccttctg ccaactttat cacttaagac agactgcctg145441ggtcaccctg tttatgacaa tagaagaata aggaaaggta aagaaaagag gggaaaataa145501tgtgagatga agcaagatgt ggacttgagt tgaaagggaa agataaaatg aagcccttgt145561ttaataaaat gagactcagg attgtgaaaa gcagtgattg aagctaaggt gctgatacag145621agtggcttaa agtggagtga tagtgtggaa aggaaatagg atggagagaa gatgcaaaga145681aaaatgaaag gaaaatgaag ggaaagatgg gatggttgtt acataggagt gatgatgttt145741atccaaagga ataagcagga aatagaataa aaaattgtag aaccaatgga aggagagtcc145801acagactctt ggtgtaggat tcaagcagca tttaaaggaa tagcttagaa gtaggtggca145861ttaatgcctt tacatttatg aaaataggta acatgatatg gggaaaaaga acaaagggct145921gatttagaat gtaaatctat aaaaatgaat aaggccaact ttggaaatat agaacagttg145981gcttcaactt tggtgtctgt aaagagagtg tgaatgtgtg atttatagta gcctagatga146041ttccagggct ctctaggtac tggcatcaga cccgaggcat gtgttttcat aaggagaagc146101ggatacacta tgatcgctgt gtgtaacagg gagatcacca tttgtaatag ctgacccagt146161tcttggagcc attgcctcac agtcccacac agctgctgtg tttgagagat agcaaaaatg146221tgcaagaaag tctaagtcag tgacaatgca agtatgcaaa acctagaggc agtcagacct146281ggaatttgct cagtgatgtt tggattttat gtctctatat ttatagctat gaaacatctt146341ttattactga gattcctgta agaaaaatca cactaagctg gtacctaacc tccttttcct146401caatgaaaca gttattttca tgaggtaatt tttttttcat aaaaaggggt ttcttctgtc146461cacggagtca gtaagaacta cccctctctt ctactcttga ttcagacctg cctcttctca146521aagccatgcc tgaggtttgt agcctgaggt ttccatggtg acagtaaaaa gccttagctc146581tgccttgttc cagtcattga gcatctgcct ggcagaaggg gcagagatgg tgtggaggag146641acatagccat tcctttaaag tttcagaccg ggcattttca tcatttccac tcaaatctag146701ctgggggacc taggcccatg gatagaccta gttgccaggc agctgggaaa tgtagtcctt146761ggcagggcag ccatttccca gaaatacgtt tcttactgca aaaaaggaga aggggatgtt146821gatggtcagt tgtttttttt taaaaccaca tcccatatgg ccaaatacat tgagattcac146881acattaaaac cattgtgatg gctgaacaga acacatccag ggctagtgtc tgtccatggg146941tagctaattt cctatctctg ctttagatat actgataata acaactcatt ccatggttaa147001gagactatgg tacatccatc ctatggaatt gtatgaagct gttaaaaaga acaggcacgt147061atactacatg gagagctatc tcggataaat taagtgacaa aagtatgatg cagaatgtgt147121atgtttataa atgtgtagac aatttcagga tgatgctaaa gaaactggta atgatagttg147181cttctgggca aggggaacag tatgactgag gacaggactg ggaaacaagc ttactttctg147241ttgaatgcct atttgtatct ttttaatttt ggctcatgta cagatgtcac ctattataaa147301aaatttgctt ttcactcttt agctgcaatg atggtatctg aaagtaccag agatgtcttt147361cctgcctgag gttacctatc tcaatcttcc ctacgattct gtattcaggg tatctttatg147421accatctgct accacctcct ggaatgcagc tgaaatgaca gaaRtgttcc gagcactttt147481tttttttttt tttttttttg agatgaagtc tcgttctgtc acccaggctg gagtgcagtg147541gcgtgatctc agctcactgc aagctccgtc tcccgggttc atgcccattc tcctgcctcg147601gcctcccaaa tagctgggga ctacaggcac ctgccaccac gccctgctaa ttttttgtat147661ttttagtaga gacggggttt caccgtgtta gccaggatgg tctcaatctc ctgacctcat147721gatccacccg cctcagcctc ccaaagtgct gggattacag gcatgagcca ctgcgcccag147781ccccgagcac tcttttaagc tctgatgcgt tcttcagatt ccatttcagt gcctttccaa147841cactgaggag tagttctgga ggacctccca aagattacat tgtgagatgt atagttgggt147901aaaacgtcat ggtaagggct ctgtcttaac ttggaagtgg ctgtgtttat tgagtggcga147961attgagaagt ctttcctcgg ttttctataa atgaatattt gtttctgtaa cttaagaagt148021aggtaaactc cttgggagta agagatgttc ctaataccaa acttaccttt caggttctct148081aaaagatgaa tttggccaca aaagcagctc tgaccttata tatttcctta ctgtattttt148141cctgtgagtg ctacttgttt cttttaaaat ctggtaggac tatgggattc ctgacccacc148201ccaaaccttt gggcccacag tgcacagcta tccaagggtt caatttggca gtctgggcag148261tttttttgtt ttgttttgtt tttttgtttt ttttaaatga aagcgttatt gagatgtaac148321ccatatacta taaaattcac ctttttaaag tgcacaattc agtgatattt agtatattca148381tagtgttata tataactatc agcactatgt aatttcagaa tattttcatc ccccYcaaaa148441gaaaccccat acctattagc aatcattccc attcccccct tttcttcctc tatcccttgg148501caaccactaa cttactttct gtttctatgg atttgcccat tccggaattc tgtattaata148561gaatcatata atatgtggtc ttttgtgact ggcttcattc acttagtata atgttctcaa148621ggttcatcca tgttgtagca tgaatcaata cttcattcct tttatggctg aataatcttt148681cattgtatgg ataaaccaca ttttggatat ttgggttgtt tccacttttt gactattgtg148741aataactctg ctatagacat tcatgtccaa gtttttgtgt aaacatattt tttctttctc148801ttgagtatat aggtagggct ggaatttctg gttcatatgg taactaactc tatgttgaac148861attttgagga gctattgaag tattttccag agcagccaaa ccaatttatt tcattagcaa148921tttatgagga ttccaattgt gtttcttcac gtcctcaaaa cacttgttat tgccttttta148981attttagcca tcctagtggg catgaagtgg tattgtgatt ttgatttgca ttttctaatg149041gctaatgatg ttatacatct tttcatgtac ttattggtca tatttatgtt gtctttggag149101aatgttcaga atctttgctc atttaaaaat tgggttgtct ttttattgtt gatttgtaaa149161tgttctttat atattttgca taaaagttcc tttacagata tttgatttac aaatattttc149221cttcgttctg tgggttgtcc tttcatcttt ttgatggtgt cctttgaatc ataaaagatt149281ttaattttga tgaggtctaa tttatgtatt ttgttgcttg tgcttttgct gtcatatcta149341aaaaaccatt gcttaactca aggtcacaaa cactcctctt ttcttctaaa agtcttatag149401ttttaggtct tacatttacg tcttcgttct gttttgagtt aattttttta tatggcatga149461gttaggagtc caaattcatt tttctgcatg tggatataca gttgtcccag taccatttgt149521tgaaaagact attaattatc catttaattt tcttggcacc tttgttgaaa atcaattaac149581cataaacata aggatttatt tctggactct cagttctgtt ctgttggtct ataggtctat149641ccttatgcca gtaccacagt gtcttgataa ttgtagattt atagtaagtt ttgaaattgc149701gaagcgtgag tcctcctgct ttattctttt tcaggattgt tttggttatt ctgagtccct149761tgcatttcca catgaaattt atgatcagct tgtcaatttc tgtaaaaaca aaaaagccag149821ctgggatttt ggtatggctt gcattgaatt tgtatatcag tttggttagt attgcggttt149881taataacatt aagttgtcca atgcatgaac acaggatgtt tttccattta tttagatctt149941taagtgtctt tcaacaatgt ttcagtatac aagtcttgca tttcttttgt taaatgtatt150001cctaaaagtt ttattctttt tgatatacat tatacattat ttcttggaga tatccatgtc150061agtactgata attttgtatt tcttttatat atcatctata tgctggctgc tctgttaaaa150121accgtattat gaaaaatttc aaatttatgc aaaagttgag aacctccacc cccatattat150181tttgaagcaa atcccaaata tcatataatt gtatatataa actctttgta tatctctcta150241agagataagg tcttaaaata cttgagcaca atacctttgt catacataac aaaattaaca150301ataattcttt aatatcattt aatatttggc cagtgttcct atttcctgaa ttgtctcaag150361tttgtctttt tatactagtt tgttccattc agaattcaat cgagggctac atgcacattg150421cacttgtttt ggcatatttt aagtctctta actgataggt tcttctctca gtcttttttt150481tcctctttgc ttttcattta ttcataaaaa gtcatctacc ttgtagactt tttcacattc150541tgtattttgc tgattgtttg ttcttccatc ccatgtattt cctataaact aataggtagg150601tctaggcttg attagagtta gtttagattt ttgacaagaa tactacctaa agatgctgtg150661tacttcttac tgcaccacat taagagatct ctagtgtcta atgcctttat ttttgtggtg150721tttgattgat taatgggttt acgtagtatt ggcctgatcc atgtgttatg ataaagatcc150781cagcgacctt tcaaataatg gttttatcag ctattgatca tcattgccta catccagtgg150841tgtgtcagtg aatgtttaac aaccggccac ttgccaattt ttatggtgta aacactcctg150901ccatggctaa tttcattcct actgatgtgt tactgaatgt ggagtttgga acagatgcac150961acagttggct ctcacaagct ggcatgagct ttttccagca aaccactgct tagatccatt151021atttcattgg tagttgcaaa atggcgatac tctatttttt ctacctttat taattgggac151081tatggtttat ttaagcattt tcctattgtt ggacatttaa tttgtgacta ttatgaagct151141tgtgagctga aggagatcat aaaacataat tctcagtaca tcagtaaatt ctttatatac151201atctgaattt atagaattgt atgaaaactg aaatagactt cagtaatggt gaccagaaaa151261gtgtcttatt caaccagtat ttaatgagca tttactgggt acttggcact ttggaggatc151321ccaagaggga tatggatatg cccccaaact ctgctaatca atgtcacagt tgtgtgtgat151381tgttagaagg gtttagagtt tgggtggttg ctaactagaa tgatcagtgg atttgtgggc151441agggggtgca cagggctggt gagctaaact ttgaaggata tgacttttca gtcaaatcct151501attctttttt gacatctgtc ttgatttact tcttatgact agtttcctgg gtgtctacta151561tagtagcctc aaatctggct tgcttgcttt gttctttcct tcataaagca acaggattaa151621ttttctgcag cacacctatg atcacttccc tactcaaaac tcttcaggac ctactatgta151681atcaccaaat ttctggtgga actgaacccg aagccatgtg attatattag ctcagtaagg151741gacacagtac tgtggcctgg agtgcctggg gaggagtgta ttacaggaag gaagtagaga151801ctagaagatt caagttggct ggctggccaa gaaaaatgga gactggtgga gaaggagttt151861gtagacggtg tctttgggaa gtcttaggag aacatcccat tagggtggta gggttaagcc151921agattgcaaa agattgtggc ataagcaagg aactttggtg tggagaagtt gtcatcacaa151981agtcattttg aaatgtcgaa ggtttagcag ttgttctgac tgtatgtcac attttaaaaa152041ataatggtag tggccaggtg cagtggctca tgcctgtaat ctcagcaccc tgggaggccg152101aggtgggtgg atcacctgag gtcaggggtt caaaaccagc ctggccaaca tggagaaacc152161ccatctctac taaaaataca aaaattagcc aggcgtggtg atgtgtgcct ctaatcccag152221ctattcggga ggcttaggct tgagcctggg aggtggaggt ttcagtgagc cgagatcgtg152281ccactgcact gcagcctggg caaggacaga gtgagactct gtcgatagat agatagatag152341atagatagat agatagatag atagatagat agatagataa agatacataa gatagataga152401tagaggtaga agagagccag ttccaagatg atagactgtt tttatagatt gccttcctcc152461taacacacat ttgtattctc tctctctccc ccaccgcatt attcagcgct aatgaaagtg152521tgatggaaac agctgctagt gaatgaatac taactaatcc tgaaaaaaag atgtgtactt152581cttgaaattt tagtttgatt tacctgaaaa catatacaga aacacaacta gtgatttcaa152641ccaaccctag agttttaata gtaaaatgct taggaggaat gcatttgtgt aaatgtaata152701tgagagagga aaccagacac ccatctttga ccaagattga atctcatgca tttttgtcac152761ttaagtctgc ccatgtctcc atccaggacc tagttaaccc ttattgcgta tcctatggta152821caggaaaaat gagttattag taaacatgga taaattttag aatcatgaag tgtcaatgct152881aaaagagaac ttgaatgtca tcttggccaa ctctcacata ttaccaacaa ggagactaag152941atccaggcaa tataatggct ttgttctcca tcagaatctt tctttccttt tttttttttt153001tttttaagat ggagtctctc tctgttgccc aggctggagt gcagtggcgt ggtctcggct153061cactgcaagc tctgcaggtt cacgctgttc tcctgcctca gcctcccgag tagctgggac153121tacaggtgcc caccaccacg cctggctaat tttttgtatt tttagtagag acggggtttc153181accatgttag ctaggatggt ctcaatctcc tgacctcgtg atccacctgc ctcggcctcc153241caaaatgctt ggattacagg tgtgagccac cacatcctgc ccagaatctt tcgaagttgg153301gaaatctttg tttagatctt tgttgcagca gatttggtat caaagaaagc tatctacatt153361aaattaattt ggagggcaca ccctactact tttctacggg caataacact aattaatatc153421taaagttaaa ggatgtatta aagaatgaat atggtaacct ttgcaaggca ctcttcaatg153481tgtttttttc tgtgtttcct tctatgtgat ttaaaatgtt agtagatttc tctgaggtac153541ttcagggaga taggaaagga gcttttgccc teatagatac ttggaaataa tatatctatt153601ttcttttttt aatttcaaaa atgaaattat tattaaaagc tctaaatata gcaagaatat153661aagacaaata gtatactgaa catccagctt caacaattaa tcaactgtgt cccatcttgt153721ttcatatctg tcttccccca cattattaat tatacttttt tctaccctta gcttaaaaaa153781aactttattg aggtatatag atcataaagt ttatccattt taagtgtaaa atttaatgat153841ttctagtaaa tttatcaaat agtgcaactt tcaccataat ccagttttag gtcattttca153901gcactccaat aatactgcac atacctgttt acagttaatc cccatgccca gccccaggca153961accactaacc tattttgtgt ttctgcagat tttccttttc tgaaYatttc atataaccga154021atatacaata tgaggtcttt tctgtctgac tttgtttact tagcatgatg tttttgaggt154081tcattcatgt tgtatcatgt atcagtactt aattcctttt tattgctgaa tagtattcca154141ttatatagct atatcacatt ttgtttatcc agtcttcagt ttgacagaca tttgagttgt154201ttctgctttt tgtctgttat gaataatgct gccattaata tttgaataca agtctttgtg154261tgaacatatg tttcatttct cttagctaga tatatagggg tggaattgct gggtatgtgg154321taaatttatg tttaactttt taagaaatgg acaaattgtt ttctaaagta tctgcaccat154381tttacattca catcagaaat gtatgagggt tactatttct ttatatcctc acccacactt154441attattatct ttttgattat cctacatcct agtgtgtgtg aagtgatatc tcactgtggt154501tttgatttgt attcccctga tgattaatga tgttgagcat cttttcatgt gcttattagt154561tattcctgta ttttatttga tgagatattg tgttagtctg ttttcacgct gctgataaag154621acatacccaa aactgggcaa tttacaaaag aaaggtttag ttggacttac agttccacgt154681ggccggggaa gcctcacaat catggcagaa ggctaaaggc atgtctcaca tggtggcaga154741caggagaaga gagcttgtac agggaaattc ccatttttaa aaccatcaga tcttgtgaga154801cccattcact atcacgagaa caccatggga aagacctgcc cccatggttc aatcatctcc154861cactgggtcc ttcccacaac acatgggaat tatgggagat acaagatgag atttgggtgg154921ggacacagag ccaaaccatg tcagatgtct actcaaatca tgctcattaa aaaattgggt154981gtttgtctta ttgttgagtt gtatattctg taattcttta tatattctgg atacaaatcc155041tttatcggat agatgatttg caaatatttt ccacagtctg tgacttgtct ttttgttttc155101ttaatgatat gttttgaagt acagaagttt tgagttttga caaagtgtaa tgttaaattt155161tttccttttg gattgtgctt ttgatgtcat atctgagaag tctgcctaac ccaaggtctt155221gaagattttt tcttgttgta aaatttttat agttttagct cttacgttaa gatcttgaat155281ccattttgat ttaatttttg tataaggtgt gaggtaagga tctaagttaa tctcttcgta155341tatgaatatc cagttatccc agcactattt gtcaaaaaat actctccttt tcctgttgcc155401ttggcacctt tgttgagaat cacttgtctg ttaaatataa gggcttattt ttggacactg155461cattctgttc tgttgattta tatgtttatt tttatgctag cactatactg aatgtttttt155521aacacagggg agtcatacat aatcttgcaa agaggtaatc ccacgactca gaagtcttta155581agtatttgct acaatggcat attttttaga tagtatatag tagcttattt acttcatttg155641tggtttgtag catgttcaga aggatccttt gagctccttg aggttgcagt acttctttgc155701tttgaagcct tctgtcatgt ttattccctg aatgtagaat accctctgtc tagctgttta155761tcctccatct ctgcattttt ctgctctacc tgccctgtag gttcaaaatg tggtactgat155821tactgctcga tagtagcctt gtgtccttgg caaggtcatt ttatttctcc aggtcttaat155881tttctcatct gtaaagtgaa agaatcagat tttatctcac tacttttcta aactctgtcg155941ttaatatttt ccttgttgaa cagttacaag gatgtgtaag ttaatctctt ttaactggtg156001aaaaagtata ttctgttcca tagagaatta aatgagcttg tatttcgggg ggtggtgtgt156061ggtcatcagg ggatagaggc aggtatttct ggaaaggtgt tcttggcaaa cattttgtta156121catacactgg tgtatatgac atacatgtcc atcactgtat gctgggtgat caagagatat156181gtataaggtc atacatgctt ttatgattag aaaggagaga aagaatgaac aatttctgtt156241acaaagaaat agccattttt atgaacttgc caagaagagg aatggtcctt actgatgaaa156301tccttaaagc tttgcatgtg gctctttact tgattgatat ttaaaagtat aggtcccact156361gggaggctcc tttcaacatt ttccattcct tgagtgtttt tccccccttt ggtattcgaa156421tgcttttatt ttcttcacat aaaatattcc tagttagggg ggatgggagc ccctttccac156481agctgaagtt attccaagtt ttgttccccc tgaagaatcg gctctctgtt cctgaagctt156541tcaatggcca catctgctgc tttatttcca ctcaataagt tggaatatat tttcatttag156601ttattgatta ggggctatgt aaaaatctga aggaaattag taaatagtaa tgtttatgaa156661taacatttta cctagaaatt tctaagcgta atgaaataag aggataaaaa ttgttagagg156721ttggagtttt ggatgaaggt accaggaagc agagatccaa taaatcagta tcagaatctg156781catgtctttc aataaactta agaaattgtt ttaaacacta ggaattatta gtctgtcttc156841tgaatttagg attaaatgac agtttctttt cctgaagttt ttaatgtggt tttcaggatt156901ggggtattta ttattttcct ttggattaca cttataaaaa tttttggctg gacaaagcct156961tgctgatacc tagtatccca ctttattaac tataacactt gacgaataaa cctgcaattg157021cttgacgaat atggtcactt tttaaagtat gtgggaacaa cattcttctc tcgtcttctt157081gccttctttt ccctcccagg aaagcagttg ttactattaa caagtatatt gtcatctttt157141gcaactgcac actagattat tcagctattt tttttctttt ttttttcctt tgccttccac157201catgcgatca aagtattttt taaatttcac tttgcatgaa ggtgtacttt ttcttttgac157261tgattagggc atactggata aaatatttgg atccaatttt aagtcaagag cagactgaaa157321atattgtaag agaagatttt taaaattaaa tttaattcca attttcaacc actgtgtgag157381tacctactaa acataaggca gtgtgtttag gtgctgtggg agacaaaaag atgaattaga157441cagaattgat agaatgtagc tagattaata tctcccacca ttaaaggaaa ggaacacaga157501attataaatg caaggaagag atttttcagc agcaaccgaa caaggaaaaa tgcatgagtt157561aatgctatat atatatatat atatatatat atatatataa tttttttttt ggaaagagtg157621gcaactgagg gaaacctgag gctttgggtg cacttctgac tctgcctgct tccttgcaga157681agctgtgtgt gtgagtaggc aaatcttgag agttctcatc gagaccctag gtctagagca157741aagtgaaatg aatgagtact cgtaattttc ctttttcttg actaagtgga aatgacttca157801ggaaagagag ttggaaaagt aaataatcat agctgatatc ctgatattgt agaggggaag157861gcccagagct tcatgctgaa ctggggacaa actgaagtct gagaagttcc tccaaggcca157921ttgatagaac tccaagactt aggagaatgc actctgctgc agggttttaa aatgttgatt157981attttggtat ttccttcgta attctaaata gcatatttgt gtaacttctt gtagacttca158041atttttgcca tcatcttccc tcccccaKta tctttctcct ccacccttcc aatatagtta158101tattataatt ttggttagat taatatttag tgtttatatt attattgtta tagccatgta158161agacatatag taaattactt ttttttttta cttgcacagg tttttgtttg ccttggagtg158221aataatcaac ttgtttcttt tcaaagtact tgcccttgat tcaaccacag acgtcttcaa158281ttttccagag ttcctcttat gaaattctca cacctcaggt gttctgtcag tttcagcttt158341ttgaagcaat agaaactgat tgtcttcaat gcctggtaca tagttttcat catgggatct158401ctctacactg tcatccaagg attcccttcc tctttttccc atgttggatt tcttttatta158461ctgtatcctg tgtctttctc tttctttgtt tactttcttg ttttggtgaa gcacatcctt158521cagcagcttc cttagaaagg gtgcatggag ctcaatttga gaccctgaat gtctaaaaat158581gtatttaatt atatcatcat gcaggctgct agtcttggat atacaatcat cagttggaaa158641taagttttct ttagaatttt gatggaattg tgccaatgga ttctagcttc taatgatata158701tgaagaagtc taaaaccatt tggacatctg atcctttgta tatgacttgc tctcttctcc158761tatgccctgc tgaaaacttg taaattattt tgactctagt actttgaaat atcatgatat158821accttgtcat gagtctgttt tcttccattg tcctggatat tcagtgagtc ctttaaattt158881agaaattcat attcttcatt tctgggaaat tttattgaac catttaattg attatttttc158941ctgctgtttt ctctgtttct ctttgtggat cttttatttg gctactaccc tattttctgg159001aagatttcct cagattattt ttgacctttc tattctcttt ctgctatctt ttaatttcca159061agtattcttt ttgttttgtt ttgttttctg aatgttcttt gtaaatagca tcccattatt159121tcatcttagc aaatagcatt tcttatgtct tactgagact ttctctggtg actctagtta159181aaatttcatc tgtcactctc taccccattg tctcttttct cagtagaata tgagcttcat159241aagggtatag cttttttttt ccctgttttt cccatggata ttttcccaat accttgaaca159301atacctccca catagttagt gtttaaatgt ttctggaata aacttaatga tcattctgat159361gatattaata atatatattt taacgttttc tcctagtaaa ggtctagtct gtttctttga159421ggtcatttct tcctgttttt attgatccct accttaggtt agaacatttt ctgcagatat159481ctggtaatac ttagtcatcc acttatattt aataatgaag gataaaaagt tgcttggaat159541atatgagcaa attgatggaa gttgccagct attggcttca tggtagaatg gtctggccag159601gccatcctgt tggagatccc ttgatatcaa ctctttagat gtctcttcct ggctgatctg159661attctctgga acacattctc ctaatctcta gaaggaagca gcagtctggg agcagaaggg159721aagaaggctg gaagaatctc agcatccaaa gtgcatattt ttactgaatc tgtctgtttt159781cagcatggcc ctcatgcctt catctgggcc tggtgccttc cctgaaaaga aaactatatc159841tagcattttg ctggagtgag aaacatgctg aaactgtgca gaatgtggga gaaaaaactg159901ctcctaaact gcttcaatca gtcttttttt ttttttttaa tgaggatgga acataatcct159961atagatttta agatagttta aagaactatt tgttgatcac ttactgtatt ttgagttatc160021aggtgagaca gatataaggc atagctacta cagcagaggg gactgtaaga gtagggcccc160081tgaagccaaa aacagcatga agaatagcta gaaccagatc ctgaagggtc ttgtcaaagt160141atgccctttt cctgaagggt aggaaataaa tccactatta gttcattgct ttaggttttt160201attgctttcc tcattttacc cctacattca gaggtgtttg atgccaccaa ttcctgagaa160261ctggggggct tctgtggtac aaattKgtta ctccctgctt tctcagatct gtctagtcag160321ctattagttg ctcatctgtt ttcccagttc caaatttaaa caagagcatt taaacagttc160381tcttcatttt tgtgttaatg agttaagagt gagattagtt acatgtattt gaactgctgt160441ctatactcaa aagtctttgg ataggacatt taataaaacg ttgggtcata cacaagaaac160501aaggaatttt acacagtatc aagccaagat tttagtttaa agtggtccca ggttgatagt160561accctcagat ttctggcaga agcgatagag ggaagaggaa ttcactttcc acatgcctca160621aagttttcct gtcggtaaag ttccaaggaa tttgagttta gagtcacagg ttgctaagca160681cactaggaaa tgaagcacaa tgagcaagaa caaacaggaa atacaaagcc ctcagatact160741gaaattatat aaatatgttg aaagaaaaga ggaagaggcc gggcgcggtg gctcacgcct160801gtaatcccag cactttggga ggccgaggcg ggcggatcac gaggtcagga gatcgagacc160861atcccggcta aaacggtgaa accccgtctc tactaaaaat acaaaaaaat tagccgggcg160921tagtggcggg cgcctgtcgt cccagctact tgggaggctg aggcaggaga atggcgtgaa160981cccgggaggc ggagcttgca gtgagccgag atcccgccac tgcactccag cctgggcgac161041agagcgagac tccgtctcaa aaaaaaaaaa aaaaaaaaaa agaaaagaaa agaggaagaa161101aatgaatggc caagaaatta taaaaatgaa aaaaacattt tcaaaaaaga ccaaatagaa161161cttctagaaa agaacaattc tgtaacataa actcgaagac tggattttgt tagacattaa161221gaacgactaa agagggaatt ggttacttgg acagtagatc gtagaaaatt gtctagaaca161281cagtacagtt agaaagctct tgaaggagag gggagagaaa aatggtagaa atacaacatt161341taaagaatca ctgagaattg accaaaatta atgggttgct acatggttca gaaattccag161401tgaagctcaa gcaagtgata aaatgatggt agagaaatac atgtgcagac ataggataaa161461gtttgatatt tcagagtacc caaaacaaag aggagatctt aaaagcaacc agagagaaga161521tgggccatct aacaaaagag tgatttgact gagagctggc ttcttaatag caatagtaga161581aaccagaagg cagtggaatg ttaccgtcaa tgtgctggga gaaaatagct gtcaatttag161641aattctatac ccaattaaat cctctttcaa gaggggaaaa atgacttttc aatcaaataa161701actaagtttg ctactaatag acttctcaat taaggaaatt ctaaaagaca tactttagac161761atcataaaat aaactgtaat ggaaattagg aagcttttag cactgagtga taatatcaca161821aataaaaacc tatgatattc tactaaagag gtacttggaa aaaatttata accctaaatg161881cttacattag aaaataaaag ctacaaatta gtgaactaag caaccaattt aagatagaaa161941aagaacaaga tatgcccaaa gaaagtaaat ggaagaagat aagcagcatt aataaaacag162001aaagcaaaca tacaatacag aggctcaaaa aagttaaaag ttggtttttt gaaaagagta162061acaaaactga caaatgccag ctgagattag tgaaggaaaa ctggagtatg aggaatggga162121aaagacaatt acacatgcag agattaaaat aataataaga ggggggtatg gacaccatta162181tgctaataaa tttgaaattt taggcaatga ataaatgcct agaaaaatat catttatcag162241aatcgactta agaattagaa agtctgaaaa gtcctataac aattcaagaa gcagaatcat162301agtttaaaat cttgtttaaa gaaaactccg ggctcaggtg gctctcctgg gaaaattttt162361atttgatatt caagaaaaaa tttccaatct tatacaacct atttaagagt ataaacaggg162421catggtaggg gtttgggggt ggtgctctcc acctgaatga agcataatag aggaccatat162481gagaaaggaa atcacttgca ggtctcactc atgaagatag ttgcaaaaat cctaaacaaa162541atatcagcta accaaatcta gcagatataa aaaacttacc aagttgagat tatcccaggt162601ctgcaagggt gttttaatat tagacaatcc agaatgtaat ttaccacatt aacaaattat162661aaggaaaaaa taataccata atctcaatgg ctagagttac agctctgatt aaaatccaac162721aacaaattat cctgaaaaga agtctcaaaa taaggatagg aattttcata ataatctatt162781tttttcaggg attatatatc tatagtttta ttaagacaaa aactgagagt gtggtgtgaa162841gtttacattc aaacaaagtt ttcaaagaaa tctaacacat gcctaaaaag attttacaat162901gtagctctag atccaagtct agataatatc aagaactgat ggatctcatg actcaagaca162961gagcattttg ggtatcagtt acttcttagg atttgttaaa aaatggtttt gtgtgtgtgt163021gtgtgtgtgt gttttaaagt gaaccactgc ccagtatgaa agtttaatct cctgagacca163081aggcttttga aatcactcaa ctcttgagtc aattcagtga aacttgtgct gtcagtgact163141gaactctgcc accaatggtt tcagagttca aagctcaaaa gagaatggct ccaaaagttg163201tcccaactaa gagcatgggc cctttacttc cctcctgtct tatctcctct accaccctct163261ttcccttccc caacacctca gccagtggct aggatgaaca agctgatatt tataacttta163321ttattggaaa agaaagggtc ttctaatttc aggaattagc acctctgagg cagaatgatc163381tgcttgtact gtatactctc caataaaaga ccttccttcc tcagtcaatt tccatctcca163441aaatggcaac tctggtaact tattaactgg cttagtcctt tgtgagacta agtacattaa163501gtttaggcag ggaacaccat ggcttataga tttcagacat tcacagcttt taaagtctct163561cacagtccgt atttatggtg ccagtgacat ttttttcaaa ggtaaaatat tctggacata163621gaagcaaatc attagttttc tgttctttgc attgcttcac catgtcccct atcaggcccc163681aaattttaat caaaatgatg ctacccatct ccctttccac cctccccatt cctccctctc163741ccacataata caccagtaat agccaaaaac tacacatKtg ctacgctgta aaaatgtaga163801gttaacacta ttgggaagaa agttgtgcgt tgcggagatg ctctttgatt atctacagta163861tttttttgct ctcccacaca cccagttctg caagttttgt ccttcataaa gggccctttg163921ctttttctca gcaaagtgca gaaaaggttg ccttgaagca cttatccccc ttcccctcca163981ctgggacggg tgtgcaaact atacagcttg gaatggcttt aaaacacttg ggctgctgca164041gggcacagaa acgatactgg ctcttcaaag ggcatcttct caagctacct ctatggtgtc164101aagacaagta gaacttcttt ccttcccacc tgaaacccac agtcagatgt gatagaggct164161ggtactcagg agagttaatt cttgtcatca tcctctttgg ggtaggaatg ccacgtcagc164221ctttcctcat agaaggatat gacaacccgt gggcatttga cattgacttc cttggcaggg164281acgaggtcag cctcatcaga atttttccct ttcatcagga acatgagctc tccactggag164341tctgtagctc caataatctg ctccggctcc aaaccctgag caaagcctcg tggcttttct164401gactcttctt tcttcttctt tggtttgctc tcctctccct tatcttcaga atcagaatca164461gctttgtgct tgcctccctc tgatttgtct atctcatgtg ctgttttctg tgacggcaga164521aactcatgag gtcagggcaa tccaagttct cttctgactc ccatgtgttg tcctcatctg164581agaatccctt ccactttagg aggtactcca ctttgccctt taccaatcga cggtagagaa164641ctttttccac cacatattcc tgttcctcct cttctagcac ctcctccact ttcttcttgt164701tttgtttttt ctccatagcg tgtgtgtaaa gggtgatgct gctcagagca gtgcccagga164761gccggaaagg aatcggtgct gcgctattgg cgcattgggt cggccagggc agtggcgctc164821agaaaagcag caagggtagc tgcaaaggag ccccttgctg aaatggtgta ccgcaggcct164881cggctaatgg ccctcctctc agccgaacaa aagagccctg cactctgtgc tgcccactgc164941ccactgcctg caccgccttc ataatctatt aaagggtatc tacaacatac ctacagcaaa165001caccaacacc ataccttatg gtgagctagt aaatgctccc ctttgagatc aagagaagac165061aagaatgccc attatcacca tttaaattca acttttacag tagataattc taactaggta165121agactggaaa aataaaaaac gtggcaaagg tactgaaaag aggaaacaaa gctgctctta165181gtatttgttt aatatggttg ggcacagtgg ctcatacctg taatcccaac actttaggag165241gccgaggtgg gaggattgct tgagcccagg agtttgagac tgcagtgtta atgatcatgc165301cactgcactc cagcctgggc aacagagtga tatcctgtca agtataatat ttttgatata165361aatgtggggg cttcccccca aaaaatgctc tcagaattta atgaggaaaa taacctatgc165421caactcttgt ttttttttct ttttcttttt ctttcttttt tttttttttt gagatggagt165481ctggctctgt cacccaggct ggagtgcagt ggtgtgatct cggctcactg aaagctccac165541ctctcgggtt cacgccattc tcctgcctca gcctcccaaa tagctgggac tacaggcgcc165601cgccaccatg cctggctaat ttttttttgt ttttgtattt ttagtagaga tggggtttca165661ctgtgttagc caggatggtc tcgatctcct gacctcRtga tctgcccgcc ttggcctccc165721aaagtgctgg gattacaagc Rtgagccatc gcgccaggcc tctatgccaa ctcttacaat165781gagaSattca gaatgaacaa cggacacata cctgtgatac tgttttgttg cttgaagctc165841aattgtgaga aattaaaaca attccatatt aaatggcctc agtcttttac cctatttctt165901cttcatattt ggggcaattt ctgataaacc acaataaatg atgttgtagg tgacatgctt165961ttatctaaaa gtgtattcag aaatagaccc acacatcatc ttttacaaga gcagggcatt166021tgtctttaaa aaaaattaaa tcatacaaaa gaacttaatg agatccgaat caggatctaa166081atactgtttc taataactct caaggcagtg agtagggaca gtcagccaca gggctcacct166141ggttggttcc tctctctcag gggtcactgt cttttgttgc ctgattttct agtgccttga166201aaactgttgt ttcatatttt tttttctatg tgtttttggt tgttttaaat gggtgggtaa166261atctctctgt tacttcatct tggctggaag cacaagtcaa tttaattaaa tggctcatca166321tttcttgtat agtcctcttt tttgttaata aggagttgtt ttaacacacg ttaatacaat166381tgcttggcct ttcccttaaa gtttgcgtag atttatttta ttttatttta tgttatgtta166441tgttatgtta tgttatgtta tgttatgtta tgttatgtta ttttatttta tttatgaatg166501agacagggtc tcactctgtt gcctaggctg gagtgcagtg gcacaatcat ggctcacgga166561agcctcaact cctgggttca agtgatcctt ccgtctcagc ctcccaagta gctgggacta166621caggtgtgca ctaccacgcc tggctagttt ttgtattttt tgtagataag gggtttatcc166681atgttgccta tgccagtctc aaactcctga gctcaaggga tccgcctacc ttggcctccc166741aaagtgctag gactacagca tgaggcacca tgcctggctt gtttcatttg ttttgaaagt166801aactgttaat tttaaaggtc caatctgtga ggatctaaag agtaggagcc ctgagatgaa166861taattcctga aggctttcag cattctctcc tgctgtgtgc tttgttattg acagctgttc166921aaggtctcac cacctctgga gttttaaccc accgatgtgt ttcataaatt ttaatgaaag166981acagatagag agaaatcata gataggccaa catgattgta ccatatccat gcactggctc167041aaaatgaaaa tctctgtgat attaccctga gttacatttg tggaacacac caaactccgt167101cttctagctt gtctgatcca gctagctaag ggaaatcaaa agtaatttgc aaaatctttt167161atgttctttt atgttcttgg tacattgctc tgatttattt tttaaggcac agaaaattgt167221ccaaatgcat agtacaaatt tgcaaataaa ctacatatag ttatttggac gatggtgttt167281ttctaaggag ttttgtaaag aatgtggttt aacttgccag tgtgtaatgc taaacttatt167341tcttgtgtgt tattatgaac tttatttata atttcatagc agctgtttat tttcattgct167401aaggtacaat tcagatgttc ttggcttaac tcagtattct ctatctcaca aaatttaaaa167461ttagactgat actttgtaaa gattcttcag atgtggccag attattttag Raacagcttt167521agttatccct gataataaat attagggaga gaaaaagaaa ttgaggtctt tgaagaatcc167581ctaaatatca tttcagtcta tcactgtcag cctccctcac tgctttcaga tccattccca167641ggagcattta cagggtggcc aagcttcctc atttgggttc cagctctctt agtttctgta167701gcctactcat tgagctaatg agcagtgtag tggctgagag ctagggagaa atatgagaga167761aatcaaagaa gaccctttct gacttttgcc ctagattgtt tgaaacgaaa taacattaga167821ataatgagga attgaccacc tttcccttgc tgcgttttag gtgtcttttc tgtaccattt167881tattatgtcg tcttaatcat aacagcctac ataaatactt gagtttgggt taggatgaca167941atgaaaaagg gattagaaat aacgtgtact cttttttctt aagccttcag aaggactgta168001aaaacttaaa gatgttaaat gcaaatgcca ttgccattcc aagttaacaa gcataagagt168061tttttgttgt ttatttttgg ggggtgatcc ttaatagtat gtgtttttct tttcagctac168121caagtcagga ttgttacccc caccacctgc gctccctcca agaccttgtc catcacaggt168181aggagacata tttgatttat gtaaactgtt cggtggtagg gaagttgtcc tatcgctgtg168241cttctttgca agggttgatc gccacagatc tggtccatga gattcccaag ctgataccct168301gtgaagagag cagagagtgg tttaatttat tctcatccag ccagctcccc agcctcatgt168361ggaagagaga aagtttcYtc aataagccac caagcataca tatactagtt cactcttggc168421tgtccagggg tgcgtctgtt ttcatcccca gagaagggta gaagaagggg cctctctcta168481attggaggtg gagacatggt ttttctgtat agtatgtgca atctgggtgc ctatattttc168541gtcagtactt taggagctat gttaattcta cataaatgaa aaataaatgg ttatcaacat168601ggggttagtg ccgtggtttt ggcctatggt gctttctgta ggatggtatt tttcctcaga168661acaaaaagct gagatagcag gagggggaga acatgtcacc aagatctgtt tcacccctaa168721attaattctg ccttatttac atgtgccact atggccttat tcatcaacac tttgcagagc168781tgaaaatgag aagcaaacag tggagtgtgg tatatggtgc tacagatctt tccttttgtt168841aattgctgct gaagaaactt tctatcttta aagacccttt ctgatttttg gcagaaaatt168901ttgggaaatt tctctgtaca cagtccctgt ttctacttca ttgttctggg tggttcagag168961ctatccttcc atacggggac atctcaggac tcatgagtca ctgaaaccac ttttaacaca169021tgcttgtcct tctttcagtg aatgaaatgg tttttagcat ccactcaacc ttgagatgca169081aatactatga gatattggct tcctgcttta gtgcgaaggg taataaaaag cccatttctt169141cctgacctgc atataaacca gcgagaactc ctttccagga tgtaacattg gccttaagag169201atattggttg aaccgtcaac aagtatttat tgagtccctg ctgtatatct aatgccaaac169261cttagttttg ccctaagcct ttcattggct gtaatttctt ctcattcaaa ttattagctg169321tgtctgttga gcatctccaa taaatcaggc attgtgctaa gtgctttcag tgcattattt169381aatcctcaca aaacccctct gaggtagata caggtactaa tattatccct tttttttttt169441taacagataa ggaagttgaa gtatatgggg gctaagcaag ttacccaagg tggcatagct169501agtaaatagt agagccagct cccaaattgc aatctgtctc ctaaaggctc ctcctgtcac169561catcactgga gtctgagtgt ctatgtgtac agccagacag cagatccatt acacttgcaa169621tggaggcttt catcacaggc ccctaaggag gaccttgatg agatgaggct cctgctccac169681accaacagca aacctggact tggtcacaac aaaatgagca ggcatttgct agctgctagt169741tcaagcctgg ggcttgagag ccacctggtt gaatgcatgg aagacctgtc tgaattgact169801tttgtaacca accttctagc tttctcttcc tgcacaaatg ccagtagtta ctacccggtc169861ccaaaatatg ctgctgctat cagcagtctt catctgagct gcttgaggtg ggcttcttgc169921ctgcttttgg agcttagtca ccaaagatct gactggggga tctgggaatt ctctagtcac169981tgttaatgaa tgagtctgct tctttgtcta ccagatttcc catcacaata taaacagtga170041gcccagtact cagcctatca gctgcctaac tatggagatt tcagatttca ggaacaggtg170101tgtaagaaat ggaggcagaa gatccacatt ccatttctct cccctattcc ctcctttcac170161ccactgtaag tgggacacac atattcagat atatgaagtc ctgggcaata aacacatggt170221tatatttagt ttttgtgcca tgtcaagttg gtattgaccc tatgagcgtc ttactaacca170281ctttgtatgg gacacctata tattctagtc tttcccaaac tgcttgtgat aaaacagtct170341ctgggagata ttaatagggc tctgtggtac agatttgcag actgctgccc ctctacccct170401ctgggagagt cgctgtgcaa gtgagctcac taaaggcttt ggaggcctac agtaagtaaa170461gctagtaaat tttgttaaag ccagcaattt cctagcttct gtgaccattg atatctttat170521ttataaaata cctgttagga tacaaaatca gtgtgcaaaa agcacaagca ttcctttaca170581ccaagaatag gcaagcagag agccaaatca tgaatgaact cccattcaca atcactacaa170641agagaataaa atacttagaa tacagctaac aagggatgtg aaggacctct tcagggagaa170701ctacaaacca ctgctaaagg aaattagaga ggacacaaac aaatggaaaa acattccatc170761cttatggata ggaagaatca atatcgtgaa aatggccata ctgcctgaag taactttttt170821tttttttttt gagacggagt gtcgcactat cgcccaggct ggagtgcagt ggggtgatct170881cagctcactg caagctctgc ctcccgggtg cacgccattc tcctgtctca gcctcccgag170941tagctgggac tacaggtgcc tgccaccacg cccggttaat tttctgtatt ttttagtaga171001gacgggattt aactgtgtta gccgggatga tctcgatctc ctgacctcgt gatccacctg171061cctcggcctt ccaaagtgct gggattacag gcatgagcca ccatgcccgg ctgcccaaag171121taatttatag attcaatgat attcccatca aactaccatt gacattcttc acagaattag171181aaaaaactgt tttcaatttc atatggaatc aaagaagacc ccatataacc aagacaatcc171241taagccaaaa gaacaaagct ggaggcatca tgctacctga cttcaaacta tactacaagg171301ctacagtaac caaaagagca tggtactggt atcaaaatag acatatagac tagtggagca171361taacagagac ctcagaaata acaccacaca tctacaacca tctgatcttt gacaaacctg171421acaaaaacaa gcaatgggga aaggatcaaa ggtcaaaata aagggctcta ttttaaagtg171481aaatcaggtt ctgataaatt tatcatttgc cctagataac attgttttaa gtttatcaag171541gttgaatatt caagtggaat gcttgttttg tgtaatgtat ttttcctttg ttaactttgg171601cagtaccttt tactgtatgt tccctagtta tagaatcggt tcatagattg ctcagagcta171661ctcatatctc tggattcatg tattatgaag aaacaattct gtttcagacc agcttgtgtt171721gattgaatct caagttttgg aagttctctt gtgaggattt cttaaatttg aaaactacta171781ttctcaagta gttagggacc ataggcacac actaccaaac ctggctttcc aggttttaag171841tatgtaagaa aaatgagttt gtacatttgc aatatgcata cagagatata taaagattgt171901ttctacatac accaaaagca tatattcttt tatcaaatga gagagatgaa gtataggttt171961tcagcataac atgcagttta gctggctcat acttattcaa gcaccactgt gatatgaaca172021tctggattaa gttatacaaa tggggccctg gagttaggaa aaggaataag ttaaaagaaa172081catcttttta tttttcatac tataggtaag atcaagaaga acctagtgac tatatacaaa172141attaagtagt ttgtgtttca cattttcctg tggcaagact agagactagg gcatggtctt172201ttggctcagc tcagaagagg tgactttttt ttgaaaaaag acttatttat agcataaata172261accaaaaagc aattaaagta aactatttca tatagagcag tgcattcttt tgtttgtttt172321tgttttgttt tcttaagacg gtctcgctgt gtcacccaga ctggagtgca gtggcatgtt172381cagggcttac tgcagccttg acctcccagg ctcaagtgat cttcccacct cagcctaaag172441tgttggggat tacaggcgtg tgctactgtg cccggcctag agcagtgtta aagtgagagt172501taaatgtatt agcccactgt ttgtatcatt ttaataaatc cagtaaattg ccatctgtag172561agtttgtgtt tattcttttt tttttttttg agatggagtt tcgctcttgt tgcccaggct172621ggagtgcaat ggcgcgatct tggctcaccg caacctccac ctccccggtt caagcgattc172681tcctgcctca gccttccgag tagctgggat aacaggcatg caccaccacg cccggctaat172741tctgtatttt tagtagagat gggggtttca ccatgttggc caggctggtc tcgaactcct172801gacctcgtga tctgcccact tcagcctccc aaagtgctgg gattacaggc atgagccacc172861gtgcccggcg agtttgtgtt tattctgtag gtcagttctc tgccaaggga ctgtttggta172921cttcagtagc taagtccttt taatagtgtt gatctcatga cagttttctt atggcaattg172981tgatagagcc acagcttcag tgtaactata ataaagaaaa gattttgggg gtgcaagaca173041ttatctttta agttttctat tagcaccaca tttgataaat tggggttccc tgcagactaa173101tgtgctcaga attttcctat ttctgccact aatttctcac cttatatgta acaaagtttt173161cctcactagc aatgtaatga gtactaaaaa tagcactgga aggtgaaagc actgtgggag173221aagcatttga tgaatacctt taagcttaga gagaccaaga agactcatgg aaggggtgag173281atgaccaatg aaattagttt taaagaaaca acagaacttt tttttttttt ttagctggct173341aaaagataca gagctttact gcagatttaa aaaaaaaaaa aaaacccaaa tgcagagaaa173401caggcttgca agaggtcctg gtatgtttga ggagctgcct gagatttgga aagccagatt173461gtagggaaaa tggctgtagt gttctggaag tggccatggg gagcaagcct cctgctcact173521tccagttcca gttctggtag ggatgtggga gagaaacagc ccctaccaaa ggagttctgt173581ctggggagca gaaactgacg ggaggacagg gttttagcct agtggttctc acccttggct173641gcacagtgga atcatctggg ggagctttga acgtgctgag gcctggagcc cctcccagag173701attttggcat ggggtgctga ggtccggcca ctgggaggtt taaaaacctc tccaggtgat173761tctgaagtgc agccagagtt gagaagcact gaattagggc aagaggaagg aatgctcaga173821gaagaggttg agaatatagg gaattttatt gatgagttct tatagggcac aatgaaaggg173881gttctggagt tggggaggca tgaggtcagc ttaggggatg tactcagacc ctatggggac173941aagaactcag atcacaaggg gcctgggagt caagggattc ttgtggtgac tgatgtgaac174001agggctagag gatgtgacag gaaggtcctg ctggaagaac tagtcactgt gtgcctcagg174061tcctgtgtgt actactgtga gaggctagcc atgaacacag atggcctcaa gaggaaacca174121gaacaactgt ggtccattga gttaattctt catctaactt cataatctgg ctagaatcag174181gtcctgaaag tgatattgaa cccaatagga atatggcagt catgctactc tttttcaaag174241actaattttt tagacacaac tttttataaa atactacatt tgttttattt ggaagtaaaa174301actgcagctc acttaaactt aaatggcagt tcgtaggccc tgaatttgtg aaatattcaa174361caccatttta aatactctat ccttttaatt aatatcctgt catttaaatt attctttcca174421caagttaata cacagttggt gattcaaaat agtaaattct tctgggagaa attaagagtt174481ttagaccaca catacttttt tggggcaggg gaaggaattc ctggtgatta tgtaagtctc174541ttacagggct atctaatcca tttttaagtg ttaattttat taaagcataa catacatact174601gaatctattt ttaatacata ctaaatcaat ttttaagata gtaaaatgaa gtataggttg174661agtacccttt gtctgaagtg cttgggacca gaagtggttt agatttcact tttttttttt174721tttttttggg aatatctacg ttacacttac ctagttgacc atctcaaatc tgaaaatcca174781aaatcctaaa tgctccagtg agcatttcct ttgagtgtca tgttggtact caaaaagttt174841tgggaggccg aggtgggtgg atcacctgag gtcagaagtt cgagaccagc ctggccaaca174901tggtgaaacc tcatctctac tcaaaaaaaa aaaaacaaaa attagccggg cggggtggca174961ggcgcctgta gtcccagcta ctcaggaggc tgaggcagga gaatctcttg aacctgcaag175021gcggaggttg cagtgagcca agaccacgac attgcactcc agcctgggca acaagagtga175081aactctgtct caaacaaaca aacaaacaaa caaacaataa aaaaagaaga ggccaggctt175141agtggctcac gcctgtaatc ccaacacttt gggaggctga agtgggcaga tcacctaagg175201tcaggagttc gagaccagcc tgaccaacat ggtgaaaccc catctctact aaaaatacaa175261aaattagcct ggcgtgatgg tacgtgcctg taatcccagc tactcgggag gctgaggcag175321gagaattgct tgaacccagg aggttgaggt tgcagggagc cgagattgag ccactgcact175381ccagcgtggg tgacagagca agactctgtc tcaacaacaa caacagaagt tttgggtttt175441ggagcatttt ggattttgta ttttYggatt ttggatactt aacctgtatt atattgttgt175501cagctttcaa atatctataa gaaaatagca agaaacatta tttaatgtga acaatagaga175561aatgtcactg cttttaaaat tgcatgtttt ttttatcaaa ttaaaactcc gtgaataaac175621accaaacatc atgaaatctc ttgctgtaat atattcaaat gacctcatgt aatatataag175681tcattgaatc accttaagat actgcctgtt aaagcatatt tatgaacttt caaaggtata175741tacaacatcc taatttttac tcatcaaaat acgaatttaa tatgtatcaa gatagtttct175801attcatgcat gtattagaaa atcagaaatt attccaaagt gaatttataa acgtatatca175861cttaagatgc caccttcatt ttcttccaaa gtgttacttt gttctgtcct acagtgtatg175921ctgtagatta gctgaaaatc acaacatgtt aatgagtatt tcagtgatta aggaaagaag175981atataaacca aggctgtacc ttcatttgtg atgatgaaag aacatactga aagctgtacc176041acactgtgct gaatgtagta atatgaatgt gtctgtctgt ggtcttatcc ttagtcctgc176101tactgtgtag ctctctggct ttgtgggaat taagtaagtt agtctcatgg aectatcgec176161ccttctataa agtaagaaga ttggactaga gggatcctac tagtgctaag atgtctgatt176221ctattttgta ttctctgttc aagaattttt tttgtaatat agataaaact tcatgaaaca176281cagttgaggg acaagatagg ttggactaat tttctctgct cctccctgct aagtacaact176341aaaaccccca gaaatactat aatggaaaga ggattttttt tttttgagac agggtctcgc176401tctgttgtcc aggctggagt gcagtggtgt gatcatggct tactgcaccc tcaacctcct176461gggctgaagg gactcagcct cctgRgtagc tgggactata ggtccttgct gccacaccga176521gctaatgttt ttattttttg tagagacagg gtcttcctat attgcccagg ctggtctcaa176581attcctgagc tcaagtaatc ctcccacctc agcctcccaa agccttggga ttacaggtgt176641gagccatcat gcctggccag aacagacttt gaaaggttta aagaggaaag aggactggca176701tgacagtgaa ctagagactt caagtagaca gaagaccaat aaggaaatag aaagcttgaa176761caataccatg gttcaacttg atataacata actatagaac acccagcatc agcacaatag176821acatttttct caagtgcact tggaatattc ttcaggatag accatatgtt aggccacaaa176881acaagtctca ataaatttca aaagattgaa attatattaa acatcttctc tgatgtcaat176941ggaatgaaac tagaaatcaa taacagaagt aaaatggaag attcactaat atgtagaaat177001taacatactc ttaaaaacac aaagaagaaa ttataagtga aattagaaaa taccttgaga177061tgagcgaaaa caaaatcata tcaaaaccta tgggatgcag tgatagcagt actcggcggg177121aaagttttag ctataaacac ttaacattaa aaaagaagag atccaaaatc aacagttagc177181tttacacctt aagaaactgg aagaaggctg ggcacagtgg ctcacgcctg taatcccagc177241actttgggaa accaagacgg gtggatcacg aggtcaggag atcaagacca tcctggctaa177301cacgggaaac tccgtctcta ctaaaaatac aaaaaaaatg ttattctggg catggtggcg177361ggcgcctgta gtcccagtta ctcgggaggc tgaggcagga gaatggtgtg aacccaggag177421gcggagcttg cagtgagcca agattgcacc actgcactcc agcccgggcg acagagcaag177481actctgtctc aaaaaaaaaa aaaaaaaaaa aaaaagaaac tggaagaaaa agagcacact177541agatccaaag ctagcaggac agaaataata aagagtagag tggagataaa caaaatagag177601aacagaaaaa caRcagagaa aatagttgaa aagttgattt tttgcaaaga tcaacgaaat177661tgacaaaact ttagctagac tgaccaagag tataattaag catgtgttaa cctctggtca177721aaataaaggg ctctatttta aagtgaaatc aggttctgat aaatttatca tttgccctag177781ataacattgt tttaaattta tcgaggttga atattcaagt ggaatgcttg ttttgtgtaa177841tgtatttttc ctttgttaac tttggcagta ccttttactg tatgttccct agttgtagaa177901tcggttcata gattgctcag agctactcat atctctagat tcatgtatca tgaagaaaca177961attctgtttc ggaccaacct gtgttgattg aatctcaaga gttttggaag ttctcttatg178021aggatttctt aaatttgaaa actattctca aatagttcgc actgaatgac aaattcaaaa178081ggcaaaggaa atggccatag tgataagtgc aaggagaaga gagggtgaga attagtaatg178141aaataccagc attggaatgg aggagaaaag tgtaagcctg tcccccttcc ttctttttga178201aagaagtRct cagcattttg ctctttatca ataagtgtct ttagaattaa tcctagtaac178261taattcaaaa gaaaRgaaaa gttctatgat gacatttatt ttagtattat aaaaataaga178321acaatacaga ccaccttaat atctaggaag aaggcactgg tcaagggtaa tgtatatctg178381cctgattaga atattaggca tctattaaaa tgattacatg acaaaacagt agaaaagagg178441ggtaacagct gaatacagag gtgtttcctg attgtaccac ataagtatat ggtagacctt178501tgatggtaag gttttctcat ctctgaggat gaccttgaag gtccatggtg catggtaatt178561ggcccctttc cagtggtcgg atttcgaatt acttttacat tagccactgt tccctcccca178621gtagggtttt gagattctta gctgcttagt gaatttctac taactctttt taggggtcta178681aagggatcgc ttaggtgact catttataat ttacctatct tgtcttataa cttgttttta178741taagttttta tattttggga gatgatgtat gtgtagtatt tatgtgaatt gggatatctg178801tgaaggtctc cagatggttc agtcctaaat gtcaagtgcc tactgttaaa tacttggaga178861agactgctaa gtaattggta gaagtttaaa tggctgtttt ggtggggaag ttatgcagtt178921tttcttttaa aaattttgtc ttaaaattaa ggattgggga aaagaaaaaa aagtcctctg178981atatgacaac tagaataaaa agtcatttga agatttacca gaataaaatg acctttggaa179041actactttta gagaccaget ctcagaaata gtgataagac atatattgca taattacatt179101gttacgatga ctctagcaat tgtgaggttc aaagaaatag tgtactgaag tattatcatc179161tagtcatttt ttaaatctat gttctttgga tagtaaactc ttattgccct cactcttaaa179221tagtaattct gctgtgtaca gaattttaga ctgaaagtaa tttttcctca gtattttgaa179281tgcattacct cctcttctag catttattgc tgccactgct ccggattgta ctgtcaatct179341attttttatt cctttggtgc agctttgtct ttcctctggg attgctttta aaagtttatt179401tttactttct gaaggggtgg gttgcccctc cacacctgtg ggtgtttctc gttaggtgga179461acgtgagact tggaaaagaa aaagacacag agacaaagta tagagaaaga aataaggggg179521cccaggggac cagcgttcag catatggagg atcYtgccag cctctgagtt cccttagtat179581ttattgatca tttttgggtg tttctcagag agggggatgt ggcagggtca taggataata179641gtggagagaa ggtcagcaga taaacacgtg aacaaaggtc tctgcatcat agacaaggta179701aagaattaag tgctgtgctt tagatatgca tacacataaa catctcaatg cgttagagag179761cagtattgtt gcccgcatgt cccacctcca gccctaaggc ggttttcccc tatctcagta179821gatggaacat acaatcgggt tttataccga gacattccat tgcccaggga cgggcaggag179881acagatgcct tcctcttgtc tcaattgcaa gaggcatgcc ttcctcttat actaatcctc179941ctcagcacag accctttacg ggtgtcgagc tgggggacgg tcaggtcttt ctcttcccac180001gaggccatat ttcagactat cacatgggga gaaaccttgg acaatacctg gctttcctag180061gcagaggtcc ctgcggcctt ccgcagtgtt tgtgtccctg ggtacttgag attagggagt180121ggtgatgact cttaactagc atgctgcctt caagcatctg tttaacaaag cacatcttgc180181accgccctta atccgtttaa ccctgagttg acacagcacg tgtttcagag agcacggggt180241tgggggtaag gttatagatt aacagcatct caaggcagaa gaatttttct cagtacagaa180301caaaatggag tctcctatgt ctacttcttt cgacacagac acagtaacaa tctgatctct180361cttgcttttc cccacacttt cRgtgttctg ttttgatggc gctgcgatgg gtttaagcgt180421agatttaaat ttgtttatgc caggtgacac ttggcatgca ttttccttct ggaaaatcat180481ctttaatttt aggacattta gtgatatctc tgtgggtttt gcttctctac ttatttcttc180541tgttttcttc tggaactaac tatatttctg tcaacacatt taaaaaaaac tctagtaatg180601tgtctttcat cttttttttt tcttttttga gacagggtct tgctctgtca cccaggctgt180661gcaatgggat gatccatggc tcactgcagc ctcaacctcc tgggctgaag ctgtcctccc180721acttcagcct cctgagtagc tgggactaca ggcgcatgcc accacacctg gctaattttt180781tgtatttttt gtagagatgg ggttttttca tgttacccaa gctggtcttg aactcctggg180841cccaagcagt cctcccatct caacctccca aagtgctggg atgtacagat gtgagcaccg180901caccttgtca ctttcatcta tttatgttgt gatctttctg ggcttgctct ggatgaaatc180961ttcatcttcc aaattaaaaa tcctctgcat aactatatag cctagaattt ttcctattct181021gttattaagc ctctatcact attctttgat tctaggtttt ataatccatt cttttttata181081acaacctgtt cttgttttac ttttccctat ttttgtttta tacccatcta ttctttttta181141ggatctattc tccactttta actttgagga ttctaaaata cttactttaa agttattttt181201aacttgttct atttttgttt ttgtcaggag cgaatttgcc ttttttattt ggctgttcta181261ccatgctttt tggaattttg ttttgtaggc ttattttgag ttggagttca atctcattct181321cagttctctg agctcctcac tttctgccag ttcagtggct gtctctacca actattcctg181381gcatccatgg cctccagtca agagctgatt cttatacttg aaatacagta cttccatctg181441gtgcaggtgt caggggtctt gtagatggca cccaatggtg cttagcccag aaccaggctt181501cagtgttata tctgcttcag ttcctttcct aagtaggcaa cccagagctt ctccacaggc181561ttcatcttgg ttttgcagct tcacaggtag caaagtgata ccttagcctc tggcatcagg181621gaatgagctt ggtaccagtc ccctgcctca tgcgggacat ttatttattt ttatttttta181681ttattattat actttaagtt ttagggtaca tgtgcacaac gtgcaggttt gttacatatg181741tatacatgtg ccatgttggt gtgctgcacc cattaactcg tcatttagca ttaggtatat181801ctcctaatgc tatccctccc ctcttccccc accccacaac agtccccagt gtgtgatgtt181861ccccttcctg tgtccatgtg ttctcattgt tcaattccca cctatgagtg agaacatgca181921gtgtttggtt ttttgtcctt gcgatagttt gctgagaatg atagttaaca gcttcatcca181981tgtccctaca aaggacatga actcatcatt ttttatggct gcatagtatt ccatggtgta182041tatgtgccac attttcttaa tccagtctat cattgttgga catttgggtt ggttccaagt182101ctttgctatt gtgaatagtg ccgcaataaa catacgtgtg catgtgtctt tatagcagca182161tgatttataa tcctctgggt atatacccag taatgggatg actgggtcaa ctggtatttc182221tagttctaga tccctgagga atcgccacaa tgacttccac aatggttgaa ctagtttaca182281gtcccaccaa cagtgtaaaa gtgttcctat ttctccacat cctctccagc acctgttgtt182341tcctgacttt tttttttttt tttttttttt ttttttgaga cggagtctcg ctctgtcgcc182401caggctggag tgcagtggcg ggatctcggc tcactgcaag ctccgcctcc cgggttcacg182461ccattctcct gcctcagcct cccaagtagc tgggactaca ggcgccYgcc actacgcccg182521gctaattttt tgtattttta gtagagacgg ggtttcacca ttttagccgg gatggtctcg182581atctcctgac ctcgtgatcc gcccgcctcg gcctcccaaa gtgctgggat tacaggcgtg182641agccaccgcg cccggccgtt tcctgacttt ttaatgattc atgtgggaca tttaaatccc182701tgttattagg aactctcagc tgcctctgtc tgcttctgca ccctctgcct gcttctgcac182761cctctgcctg acctaccact ttgtgatttg gccccacaga gacctttatc ttgcttctga182821gtgggactga gtcttttaaa ttttaacagt tcatgctaac actctcacat ctgtcattgc182881atatcatttt atgtaagggg aagttgagat tcagagagca gaagttaact acttgggcct182941cgcagttggt aagtaagcca gcctgctgta gaagaaaagg gctggaaaag gtttctgggg183001aagtgctaga aatttaataa gcatgtagag agagaaaaat cattcattat acatcacaaa183061aattaactgt agatctgcta agtaaaagtt gattaaattt ctacactctg tgcccaccaa183121taattttcta aatgtgaact ctttcctaca ttaatctcaa atgcttattt ttaaagtgcc183181ttccccagtg cagacttaca ttcatcttag ccaaaatggc cttcacattt ggtctgtggg183241acatatttga tgtagcaaaa ggccaagggt aacccttatt ctacccccta aaagggaaca183301gaaagagaac ttttgcataa taaataatta tttatgcatt tcaaaattga ttttaagagt183361acacaaacca tagaggcttt gcgattcttc atgtatttta ttaatttatc aagcacttca183421acagagtgcc cgatgctctt ctagatgctt tacaaatatt aactcattta agcctcacaa183481caaccacgtg cagtgggttc tattgctatc ctaacattgc agataagtaa acaaggcaca183541gagaacctca gcaacttgct tgaggtcagt gacagaaaga aagagttaag gctatgattc183601ctaccttggc tatttagccc cagtgtacca gctcctacta tactactatg ctgtgctgtg183661aaaccacgtc tcacaattat tttattaaca atcaaactga cacactatct ttgtcaggga183721cacattttat ttcatggaac tagttaaaaa ttagctttaa ctcacttctg gtgacttgtt183781ttctttctta ggttcttcct cattatatta tgcccatgct attgttatat aaagctcact183841ctgctggtat cattcttctg gaagatgcca taaattcatc taatcatttc atcagatgtt183901aacggaatgt gctcactgta ggcccacagt ttgctgagaa ctataggagc acaaagattg183961tgatgtgatc cttatcctgt agttccttta tcctgcgtca ccctctcaca gcagtcatgc184021ccaacactat ctattctata gtctctcatg gttcttacaa ctgagtcatt ttctgtgtgt184081ctcatttaaa cccatttctt aatgctttat tgacacaggt tgtaagcaat aaactggtca184141ccatgtttct aataactacc aatatagccc agttttcccc tggggggctc agttatggtg184201gttttctagg tttYtgagca atggaatcct catttctctt ctctgctgtc ccctatcgca184261ctgtgacttt tttttttttt tttttttttt tttgagaaag agtcttgctc cgttgcccag184321gctagagtgc agtggcgcga ttctggctcg ctgcaacctc cgcctcccag gttcaagcaa184381ttctcctgcc ttagcctccc gagtagctgg gattacaggc acatgccacc acgtccggct184441aatttttgta tttttagtag agatagggtt tcaccatgtt ggccaggctg gtctggaact184501cctgaccttg tgatccacct gccttggcct cccaaagtgt tgggattaca ggcgtgagcc184561actgtgcacg gcccacactg tgactttaag gaagcactcc tgagggggtc aggggaggag184621cagatgtgga ttgggaggat gcccaaatgt gctactgtgc taatgggaaa ggacaatgga184681ggacaaccag gggttgggct cagggaggca gcttgacaga ggatttaaga gcatgggctt184741ggggtttcaa atcctatctg cctcttactg gctttgtgac tttgggcaag ttagtctctc184801tgtgcctcaa tttcatcatc tgtaaaatgg ggatcatggt agtatccatt ttcatagggt184861tatgatgatt aaatgagtga agatctctaa agcacttaga atgatgcctg gtgccatagt184921caaccctcaa taacatttag ctgttataat tatcatcacc acaggcctag tgaaacagag184981gtggggtcta gttggcaaac acagaacaga ctccagatag ctagaaatta gttgcaacca185041gcccagaatg agtgctgaag tggctcgcta aagcacttgc agtcaaagca cattgttacc185101ctcatctgtg acctttcatg ggctgtctct ctgcctatgt caggggttgg caaactatag185161cctacaggcc aaatccagat ggtcacctgt ttctataaat aatggaaatg tgaaggagaa185221tcaattggca agaggcacgc atagcgcatc ttcaaccttt tccatcgggg aacatgagtc185281ttgagaatcc tgccatagat caaaagaatg gcaatgagaa gaaccagtgg acaggtggat185341aggcctcccc atcagccttt aaccctcctg ctcttatctc aaggatcata tgcccgtgtc185401tgtggtttct gatttttcct tgaggacctt tgaccgagat gatctgtctt tcagacttta185461ctagcaaaat tctgagcagt atcttttata caactggcag cttcaaaggc ttgcagtggg185521taggcctccc caactctgtt tcgcaagata attaaaagaa gccagactcc ttcatccttg185581aaggagagga agaagaaaac aagcctgtcc agagaaaatg gggtgaagga atttttctcc185641ccaacttcct cctaaatacc ctgctctgcc catcaaattt tgtaagtatc acccagtggt185701gtgttggtaa accttctccc ctgcggggtg ggggaaagcc ccaatttgca gccgttgttg185761ctgtctgtgt tgtgaatgct cccaccatgc tgactgcaag tggagctggg aagagatgtg185821taccatcggc tcactaacac tagcatgctc tgtggtagac agcttccctt gcctctgaat185881ccttgtgtca ggattctacS tgccaggcaa caagagcttt ttttctttat gtatcatgcc185941agtgatttta gtctagggca ctgttttagc ccaatcccag aatgtgctat tagaaatggg186001gccatatttt tgaagttgct aggggcaaaa cttccctctt gtttatgttt aacttgtgcc186061actaggtggc actttaagga cattacagga aatcggctct actggatttt taaaactttt186121ttttttttga gacggagtct cactctgtcg cccaggctgg agtgcagtgg catgatctca186181gctcactgca acctccgctt cccgggttca agcaatttag ctcactgcaa cctccacttc186241ccgggttcaa gtaattttcc tgcctcagcc ttccgagtag ctgggactac aggcatgtgc186301caccacaccc agataatttt tgtattttag tagagatggg gttttgccat gttggccagg186361ctggtctcaa actcctgacc tgagttgatc tgcccacctc ggcctcccaa agtgctagga186421ttacaggcgt gagccactgt gcctggccca aaacactttt tattgaagta taattgcata186481tagaaaagca agctgcacat ttaatgtgca actcaatttt cacaagcaga acacatgcat186541ataatcagca cccaaatcaa gaaacagaat attactagca ccccagaagc cctccttgtg186601ctgttttcca gtcgctatct cttattctta gggttaccac tattctgact tctaacagca186661taaattagtt tgacttgtgt tctactttat ataagtggaa ttatgcggta tatactctgt186721gtctggcttt ttattaaatg atgtttgtgt gatttattca tactgttgtg tgtaattgag186781atcattgctg tatagtattc cattgtatgt gcaaatataa cactgttttc tactattgat186841ggacattttt gtacttttca gatttgggca attatgaata gtgctgctgt gaacactcgt186901atccaagttg ttgagtggat gagtgttttc atttctcttg gatatatacc taagagtaga186961atttctaggt catggtgtag gcatatgttc agctttagta ggtactgcca aacagtttta187021cgaagtgttt gtactgattt acactccatt aatggtgttc ccactccagt tccatgtcct187081catcaatgct gaatttattt tatggatggt tcctaactct taacataatt tttatagctc187141cctttctggc cagctatttc taattgaaaa attttaaagc cacattattc tgatggacat187201ggctcatgca ggaagggcta tgtcaaccca aaaggggaca tttatcacag tctaagtgtg187261aactatcagt aagtcggaaa ggatgttact ctaatactcc atcacactga tacacagagc187321aaagaaacca ggaaaggaat tcagattgta agtgtgataa aatatcggtt cataggccag187381ggctctgagg cattttgaaa tttgtatata catttgttat tattgccagg cctatatatg187441tcagagattg gggcccaaca gaaagaattt ctaaaggcct cataatgaag tgcctttcaa187501ttgtgctgct ggtgggtgcc tgttttacta agggtctctt tggtttggca agaggcatgc187561atggggcatc tttcttcaac ctcatccctg gggagcgtga gtctctagag aatcctgctt187621tagatcacct tacctgtgag gctcatgttg ttgactctgc agatggtgtt ataatatggc187681tttaggtatg tgatatagag agtacagttc agacttgact cagatttctg ttccattact187741agaaattgga catagaagaa caaaacccag gtcatggtca aaatatttgt cttttagacc187801aaaataaagt gctcatagtc ctttagttgt aacagtaagt ctgtctcatg gatgttgtcc187861ataatctaat gggttccaag aatccccaag gcattctgcc atgtgactct tgaatccagc187921aactagttgc ccattatagt cactggtacc cactttcacc aaagttgttc ctgagcccct187981ctgtcagtgt tttcctgagt gtctccagtg ttggattggt ctaccgaggg gggtgcccac188041cagagatgtt cactgcacac tgcctgagat atccatgtta aagcagccgt gaaacatgaa188101agtctgaccg ggcttttatt tttaaaattt tctcccccca atattagtag cggtaatatc188161cagtggcaca gatttagact gtggatagcc tgctttgtgg gatctgctta tattctaaat188221tggagccagc attttctcag cctaggtttt catctgtttg gcaaggagga aaatgccctt188281atgttggatt ggctgctgct tgcagtagga ggaactatgc ctgggaaacc aatcataatt188341ttaatcctaa tgataatagt agctgctgtt gtcaaacagt tactgtgtaa ttgtgcttag188401cacactagac agatattttg cttaatcttc acaatattgt gagttaggtg aattatccca188461ctttgcagat gaagaaattg cagctcagag agactaagca tgtgaccagc tcacacaatg188521agcaactgca aagcctgggt ttgactccag agcctatatc ctgtaactgt tctgtgctct188581gtgggtgcag ttaccaagga atgaattact gacctcttct ccagtggaac atgaaacagt188641attgggttca ttgctggctt attccacaca ttgattagag gacctgttgt ttgctggcat188701gcagcttagg tatcaagaaa atgataggga agcaagacag atcctgccct ccaggtgctt188761accattcgtt tatcatgttc tgtctctcac aggtgacttg agtcatggat tttggtaatc188821attcttgctg agaattcagt ctcttggacc ttcttgaggc tctatttacc cctctctctg188881agcctggttg gggaatgatg gggtggaaca gtggtgtgct gggccattgg ttctacatct188941tctgtgactg ggtttagcaa atggctttct gttgtgttct tgcctagcgg agatggggga189001caggtgaaaa ctaggcacca aatgacattt agaatactga cagaatgctg gaaaggccct189061ggaggcagca gcaggattgt cttcttgtct gcctagaatt tagctgctgc ctttgtgatc189121cccattcttg gaatccacct ggatccttgg atgataaagc ctgagttcgt ttttctgacc189181tgccaatatt gcaaacctaa aaaaaaaaaa gttacctatt atccacacct tggcaaagtt189241ctacagatgg acttcttttt tttttttttt tttgagacag agtcttgctc tgtcacccag189301gctggaatgt ggagtgcagt agcgagattt cagctcactg caacctccgc ctcccgggtt189361caagtgattt tcgtgcctca gcctcctgaa tagctgggat tacaggagcg tgtcactacg189421cccagctaat ttttgtattt ttagtagaga cggggtttca tcatgttggt aggctggtct189481caaactcctg gtgtcaagtg atccacctcc caaagtgctg ggattactag cgtgagccac189541catgcccggc cagactattt tttaagtggg atttgacttt ttttttggtt gcaaatttga189601cttttaaaca aaatcttact gctgaagatt ttgcccagct gcaaatattt atctatagaa189661ccatgtggca tataccccca ttccctctat gatgaatgga tgagaaaaca attaagaagc189721ctagttagtt ttgcaagagc acctgtggcc ttgcccagac tagggccttg gagacttggt189781tttctcatct cttaaatgaa gagtttggac catcagtggt tcccagttga tctgggctgg189841ctacataaaa atcacttagg gtgcttgtta aaatataagt ttctaatgcc ccatccctaa189901gggattctga tgagggaagt ctggaatagc atctacaaat ctgaatttat aaaaagttcc189961ccagataatt atcctgctct ggttatattt gggagccact gggctagagg gtctagaaca190021aacataatta agttagggag aaacttctga tgtgcttgaa gatttgggga atacctatat190081gcctggagca ggttccatat gggcgaacta acccagttat gttgcagagg taaattggag190141ccagaggatg gaagaccttt aactggctga gggattgtgt tgtgtgccat aggctttggg190201gagccagtga agggctttga gaaaaagcat tttgattaga tggtgcttta ccagattaat190261ttctcagtgg tgtgcagggt ggatcagagt gcactagaac tgaaggctgg aaaaagaccc190321agtgaagtct ttgcagtggc ccaggtgata agtgtgacga gtacctggac tcagatggga190381ccagtgggac ctttgtaaag ctaacctgcc tcagtggaac tggaatctga gacatactgt190441tttaaactcc ttaaaaaatg cataatttat taatgtcatg ttcatctgcc taaactatta190501taattaacta tttttattgt gtttttgtcc aagaggtttt aaaggagccc tgtaaataaa190561ttcatgcaaa gaagtagttg gcagcagcaa cagctgctac cgcagggaga agggaccccc190621ttctggccag gcagcaacct cattctcacc ctgtctccat actctaagcc aaagcgagag190681gggaggggag tgcaacaggg gcccctggag ttggatctgc caaccacagt ggagaatgca190741cctccccaag ggttaggtgt ggaatatggc gtttggcatt aagtcaaata gaatattctt190801cctttgttcc tctaaaatcc agtctatagt caatgtttct caaagtatgt tccacagaac190861attagttctt caggatgtta atagaagtta catgagaata aggagatgaa accaaataaa190921tttgggaact gatgggctac gcaaaggcaa gtgacttcct ttcagaattt cagagtgtgt190981aatgttctgt gtgctcagtg aatctccaag Rgggagatag accaggcagt gtttcctaag191041cttatctgta gattcttctt ccccctaggc caggggctga caaattatgg ctcataggcc191101tcatctatcc caccactttt tttttgtaaa taaagtttat tggaacacag ttgtgcccat191161tcatcgtatt gtctgtagct gctttagcac aacaggagag ttgcagagtg gctacagagg191221tagtctggcc ctcaaagcct aagatattta ctatctgccc attacagaaa ggtttgctaa191281tccctgctct agacaattca tggaactggt gcttcctggt accccctttc agagatgcta191341ctagaagaag atatacatgg gttgttctag cactgcttta tgataaatct gcctcatttt191401attctgtttc catactgcca tcattcctgc ctcttgcttt aaactacttt atctctcaac191461catttaccca ttatccagga tccaaacctg ggttccatgt gtccttatct tttcttgcac191521tcacaacatt ccaaaaatgg ccaagggtgt cttcaatttc cagtctgggt agctcttaag191581tcagcccgaa cccattccta ccaacctggc ctacatgacc gtcatctttt ctttttcagc191641attcgaatta ggttcctttt ccccagatat gagttacctc tagtcctttt tactcactga191701atctagaata acctttgtaa aatacaaatc tagtcagatt gattccttac ttaaaatcct191761tcagtggtcc ctgtggccct caggataagc tcagatcctc aggatggatg tgtgtcagtc191821tggttgggca aacagaaact actcctggta tttcgtactg gaagagattt aatttaagga191881attagttact taagaaaagg agtagatatt tccaggcttc tgcctgctgt ggcatagaga191941agggcatgga aatgggggat ggtgttgagt gtttagcaaa tgacccagaa caaaccaggt192001ctgggaaacc atgtgttatc ccctgctgcc cttctccagc ttcagctctc ccatttttct192061cctgactcct cacccttcct ccaccctgca agtccccaaa ttctagatat atcaggatgc192121ctttttgtcc ccccagaaac ttaaaacaac tgacatttac gaactcacag tctctttggg192181tcaggaactt gggagtgact tagctgggtg gttcggggtt acatcctctc atgaggcttc192241tgtcaagatg tcttccaggg ctgcagtcat ctgaaggctt aactggagct gatgatccac192301ctctaagatg gctcactcac agggctgttg gcaggaggcc tcacttcttt gctgtatgga192361ccccttgtta ggactgcttg ggtgtcctca caatgtggca gctgactgtc tccagaacaa192421gtgatatgag agtgagagat agagatagaa acagagacaa caaggaggag catactagag192481tgctttcagt cccctctcct gggagtacgt tgcatagtgt ctcaggtcag cttccctaga192541agcagaaccc tgagatgagg attcttcaaa agtgatttgt tgaaggagtg ctctttagga192601aaaaacctag aaggaggaga gggaagaaaa gtagggaagg ggaaggagcc aagcaatgat192661atagtctcag ataaaatctg accttggcct gatccacaca ggcaggattc tggtacataa192721ttgcataata gagttaaccc tgtttgagac aggggactgg ccttttatat ccttctctaa192781gtcagtcatt ataggctgct ggaggtgggg gtgggaggta ggctcccatt agttgagaat192841aagtctctag agaagagggt ggctgtgagt cctcagtagc taacactcag agcagctggg192901ggatggtgtg ccagccaggg aaaggtgacc caggtggggc caacacagca cctactacag192961tgggcctccc cRtacccatg tcccaagcac ctaccatggt ccctgacata ccagaggtac193021gaaaaaaata tctgtgtaat tgaataaatg aatgcataga aatgcttagc ctctctggac193081ctgtgtccat ttctgggaag tcagctcagc agggatcata ttgtcccatc aaccatggga193141atggtcgaag acccaggaca gtttgccaca tcatgagtgg cagcactgtc taagtccagg193201attttcattg caaaattaga gtgaaagtat atttgccatt tgtgaataag gaagtattaa193261tattacaata ttatatattt ttaaaaggaa ctttagctca catttcagtg agctctttgc193321taaaagtaca ttttcctggt ttgtcacatc ctgactgggt acatgttgtt acacatggaa193381ccgtgagggc agaactgcca tttgttatat taattgtaat ccatatatct gtttttttcc193441cacaagaatc agtcactgga tttcagttca gaaaaccaaa actgtcggac aaagccatat193501tatcgtgttg gcagagttgt tccccatgac cccaaataca cagaggagga agagaaccac193561tgaggcccct tgttttcttg gattatcatt cttccaaagt tggtaactaa gaaaagtctc193621aagaaatagc aaagaaacct ccccacctta ttctgctttc cacatcccaa gcctaacata193681agcctcaaac tatttttatg aaggaaagct tattgtgaag agggtctttt acatgcctaa193741atctcatctt tagtttttct tatcacacag ccggttatct ttgcccatct cttttcctgg193801gataatgtct ggctctgtca aggctgtgcc ttattctggg accttcctaa tgcctgtgtt193861ccaggtccct cctgagattc gctcctccga atgcttgata gatggatcaa tttaggggat193921tgatggtcag atttagcctg acataacacc atgcagtggt ctgttaccag acatagagct193981ggaaaaccca ggcccttgca tacaagttca ggaaggtaaa tttgcaaatg aaaaacattt194041taagacacat ctcacagctg cttcctcatc ccccacccta ttgtttgtgt ttttagagat194101gtttaagaga aaaggaggca ataacaaaga cgctctgctg tgctttggaa tcctcattta194161tgttgaaaaa gggtccattt atcatctcct tggatatttc cacaaggaga ctttgctcat194221ctcaccaggg aatggagtgg tagagcaaac acttctctgt ccacctgatg atggaggaga194281accaactaac tgaaaggcag agagcctgaa attattatcc tctctctgcc tgcctgctgc194341aagatattgc tgcatacctg tggtccctgc aggaggctgc atgcgtagac ttgggccaga194401acatatatta ccttccaagc aatagatttc taggtaaaag gcaagaaggg gcctcaggtt194461caagttgcat tcttcagtgg ggttcacagg aggccctgca ttcagggggt tactgggccc194521tccttcagct gttaataaaa ctgacttttg gctcatagag cttggagtat gcgtccattt194581gtatgaacct ccagggaaag cagggtgagg gcttgtgttc cctagactcc tggcagtgaa194641tggctcctgg acagctttgc ttggttgttt cccaggtact tcaaactcaa tctgcctaaa194701gatgagttcc ccctcactcc tgcttctcct gtgtttccca tcttggggat tgaccttagt194761gttcttgtgg tagccctggc tgaaattatg gatgttggag catgggaggc cctttgtgaa194821cttacttttg ggtctttctc accattccca cacattgctc ttttagccca ggcctcagct194881gagacateac ctgctcctga gtctttgccg acccatgaga ctgggtgctt ctgtattccc194941catacttgcc ctccacagtc cactgaagca agttggcttt gtcacttccc tgttgctccc195001actagacttc aagcttctct taagcaggat ctgtatcttg ttcatccttg agtgccagtg195061ctagcacagg gcctgcctgt agtagctgct taataaatat gtcttgaatg agtaacactc195121aggcctgaga atacNtgaga ctgtatggca tagtggttat aagcttagct ttggaaatca195181aggcctgtgt tcagactctg ctatttgtgt gattttgggt gagttcctta acttcttgat195241cttcaatttc ttcaccaata agatggggat aataactact tcctagggct gtgatgatga195301attgcaatga tccatgcaaa gccctttgat caatgtatga cacatagtaa gtgttgtgtg195361catcatagta cctattagta acaacaacaa caatatagtt atcgttaatg gaagggagaa195421agctataaga actagttgga actagaagtc aggaatattg agtgcttgca tagtggattt195481agaggagatg gggaattgct tgggataaaa tgtaggaaca gatgggatta ccctgggaga195541gaatgaacag ggagggaaaa atgggtggag caactaaaga cagaaccttg agaaatccta195601catttgaggg ataagaggag tcaaaacagg aaaaagagtg aatgactaca gttaggagat195661tgcagcttct cagatgccag gggcaggagt ttaaaaaaga tagtcaaagg tgctatactg195721ggcagaagat caatgaggag aagacgagaa gaggtagcca taacgtgtta aatgggggta195781tgttagcgcc cttgggaact agccatttca gtggagtcag aggaagaagc catgctctga195841aggaggttag agaaggccac gttagggatg aagtgggcat actaagtcag tttgttgtgc195901tacaatgtgt gtttctgtaa tgaggattat ctgtgggtca gtagatgact ggtgtcagta195961tgatgtagaa gtcacattgg ttcatatgta gggttcattc tcagcacatg aatgttttgg196021catcactggc tactagtggc tgaaagcaga ctaacacacc tgaaggcaga agactggggg196081aaatggggga catagagagc tgctcatgag gcgcagtcac cctgtgttct tccacttgac196141tgattggcac ccactctgtc ttgaagtgct ttggggacat tccttgccaa tcttggtgag196201tttagtcact gattccattc cactcagcac ccttgaccct tctgataact ccttcgacca196261cagtcYccct tcctttttta agaagggcca tatttactag agtttttttt tttttttttc196321ccaaactgtg tccaggttta ttaaagatac tttttgtgaa cagtcatggt atttcaggca196381ggacatgagc gacaatcatt aacagtatac aactttcaga ctccctcctt caatggactt196441ccaaaatcag aaagccatgg taaaacccaa tgaagtcttc atttggtacc ctgaagaggg196501aaagttcaga gtgagggttg acatttcaca ttcaacgtgc tgtttaacaa cttttcatga196561gctgaccctg actttcagga aatgaaatga aaatggcaga atttatctga agatccacaa196621tctagaaaca gagcacgtgt ctttcaaggg ttctcgctgg aaagtccaga ttgccagcct196681gactggtaac cagttattgg gggccagacc ccaacaggta tctgggtttg agggagttaa196741gtgtatactg aaggcagaga gggagagggg gaaataaaga ggaatttgtt tctccacacc196801acaaggcctt tgtgcgaagg tgactgtgtg tgtcaacgtc agacagtccc tctttctggg196861agccaagaag atgtctttaa aactagaagg gaaaggtgtt ttctccacat cagtccagct196921tcatagacat tctttttctt ttttttcttt cctgagacga agtcttgctc ttgttgccca196981ggttggagtg cagtggcacg atcttggctc actgcagcct ccgcctcccg gcttcaagtg197041attctcttgc ctcagcctcc tgagtagctg ggattacagg aacccatcat cacgcctggc197101tagtttttgt acttttagta gagacggggt ttcgccatgt tggtcgggct ggtctcaaac197161tcctgacctc aggtgatccg cctgcttcgg cctcccaaag tgctgggatt acaggcatga197221gccacggtgc ctggcctcag agacattcta ttagtgacat ataccccttc tccataaaac197281aacaatgacg tgttccgtgt gctaacaaca tggcttaaaa taaaggcata aaaacaattc197341tgcattttta taaaacttaa taaaaaaata atacttcaaa ctgcacagtc accagaaaKa197401cacagttatc aaaaatgcac acacttccct tggcatcccc agcgccttca actttctgtg197461cctgctctgt tttggcctct gcattttctg cagagttatt cccctccttg ccagcatggg197521ctttttcctt tttccccttt accttctctc ccttctttgc aggggcctct ttaggcttgg197581gcttcggagg agcaggtttg gcagacaacc ttgcagatct tctctgtggt ttgtccttac197641tttggtttta tctcctttag cgtccccttc agcctttctc ttgggcatgg tggaggcgac197701tgcagtggaa tgtaggtgct ggatctgggg atgcagcagt gcgtgggctt tggttggcct197761gggggatagt tcttacctct tcttcttcac actgctcata gagtatttct ttattagaaa197821tttaacattt aaaaaattca gtgtttttaa tgatgttgat tgtttttgtt ggtgctcaag197881ttaaaatctt tgttttcaac cctgtttttc cgataagcct tgttactttt agagtttgtt197941gttgcgaagc attagttttt tagcaataca tatgttcaat tacagaagaa attcccatga198001ttccttttcc taaggttgaa caatctggga aggagggatg ggaaggttaa tagcagcatc198061tatgggaggt ttttcccaag gatcagcaga cttaagcatt cttggctgca acttggccta198121aatcagtttt cccttggtga ctatcttcct aagtgctctt tattgtatat ttaggtgatt198181gtggttttgc atttctgtcc tctctctgat ctcttgctac agtctgaaca agtgtcggag198241gccgagttac tcccacagct gagcagagcc ccatcccagg ctgcagaaag tagtccagca198301aagaaggtaa ggtcagcctg aggatgctgt cagatggcgt tgcgagtccc accccgggct198361tcttatctct attagctgat ctattgtttt aaggtttctg tgtcctgaaa tgaatgtcaa198421tgatccagat ttcctagaat gtgggttctt tgtgaagatt ggggtcctga agattcctgg198481gcttgaccct tgatgcattt tgtacatgac agtacatact caacctcctt agccccccag198541ctaaacatgg tggccttgtt tatggcaaga gctaaaaggg ctgtggggag ggacagatgt198601ctttagcaaM accccatcca cctgctaagc ctctgtgaat acctcttggg gtggggtagt198661ctctcctttt tggtctttgt accatggttt cctcttgcat cgtggcattt tcacctggca198721tctgcccagc catgccattg ggtcacctgt aatttgctca gtaagtaccc ctcactcatc198781atgctctcca ttcatctcct caccctcttc ttgtccttct cccaggctct tctttcagag198841agagtgggcc Ytgggctcag gagcacaggt gaggtagagt cctccaaaga agcagaccaa198901ctgtgtcctt ccagaataca ctgtggctct ttttcttgac tttatgtagg aattggggat198961aggagaatga tgagcatatg actaagacct ttaaaaactt ccaaagctta ggtttttttt199021tgtttttgtt tttgtttttt tttcctgaga cRgagtctcg ctttgttgcc caggctggag199081tgcagtggcg cgatctcggc tcactgcaag ctcccaggtt cacgccattc tcctgcctca199141gccttccgag tagctgggac tacaggcatc ttccagcagg ccggctaatt ttttgtattt199201ttagtagaga cggggtttca ccgtgttagc caggatggtc tcgatctcct gacctcgtga199261tccgaccgtc tcggcctccc aaagtgctgg gattacaggc atgagccact gcgcccggcc199321aaaatttagg tttaaaaggc ttcaaagtga ggcaggagcc tatcttgtct tcagttagga199381aaggccccat ggtgtgagat gcacacaggg tctgtgtgag tttcaggtga caagaagtaa199441tagacttctt gaaacattta aaaagaaaaa gaaaaaaggg aagaccactg gactctgatg199501cctgtgtccc cccagttctt gctgagccat gggtcagtaa atagagtacc ctcctgatct199561tggggttggg gctaaaggca tgtgggacac tggaaggaag agtggaggga acattgaaat199621gtttaacttt ttaattttta ttttgtttta aatccacagg atgtactgta ttctcagcca199681ccatcaaagc ccattcgtag gaaattcaga ccagaaaacc aagctacaga aaaccaagag199741ccttccactg ctgcaagtgg gccagcttct gcggcaacca tgaaaccgca tccaacagtc199801caaaagtaag tagaccacat aaacaagagt gaggtaggaa tctcgctctg agatgtcgac199861ggatatgtaa atgtgttgac atttacgtgc gcaccaacag aaggatttca tggttgtaag199921tttatcttgc tcatctctgt gatgggattc aagtctccca gtcacataga aggctttata199981agctatagga cttaacctga cctaaagggg caggcgagtc ctaggctgtg gtgtttggtt200041tggtttgatt tctttccctt cctctcacct tctgttgttt cctactcctc ttccttgtga200101gggcccaaat tttcctggca agatgaaagt attgtaagtg gcaattttgc ctctctgcct200161gtgttatctc cttttccttc agggcaaatg gctagctcct tcacccacag acacactttc200221agtaggtgtg gtatgtgatt cttttaaaat ctccgtaaag agtttctgtt gttggtcgcc200281tcagcacaga atggatacat gcagaacttc cattatattt atctaaccag ctggaacagg200341tattggaggg agggtgaaga cacctgtgga catggccagg gactgagaca atgatgctct200401ggacaaacta tccttctctg taaaactttg gctttgagaa gagccatcac ttgcttcaat200461ttctttgtag tgaaattgag ggaggtgttg aggacagaaa cctgctcctg tcacttgact200521gcaagtgagc ctgaccggtc actaacataa atgctcaaga cccatcatga ccctgctttt200581ggggtcttgc aggtctcaga tgctgctgca gattttcttc tgcttacacg ttgtagaaYg200641aaaagcagca ggttctgtgg gagatgttag caacttgtct ttgaagccaa agtcccattc200701tgaatctctg cagccctctc cttcagaaga ttttcttttt ttcaaatgga caggtccctg200761gtggtatcag tccataaagg aagtatattt tgttcatcac attcaccgag aactttgacc200821atcttccagc ccctgttaag cctggaaggt gaagaatccc tatcagggac aaagatggct200881caattttctt ccatgttcta ccccagcact ccactttaat ttcagcttta tgttcatttt200941agtcagcagt ggcttaacag tctccagagt agatgtcagg gcatctggtg gagttcatac201001agagatcttt tctttttctc ttttttataa cagctttatt gcaatataat tcacatagca201061tacagttcat gtttttaaat gtacaattca gtggttctta gtttattcac aaagtcatgc201121aaccatcacc acaatctaac ttttgtcatg cctaaaataa tatatatcca cgcccattag201181caatcactcc ccattctctc cttccccctc agcccctggc aaccacgaat ctcctttctg201241tctccatggc tttgcatatt cttgtcattt cacataaatg gaataatgaa acgtggtctt201301ttgtgactgg cttctttcac ttagcataat attttcaagg tttatccaag ttgtagcata201361aatctgaact tatcttttta tggctgaata atattccatt gtatggatag accacatttt201421attcatccat tcatcagtca atggacattt tggttatttc tgcttttttg gctcttaaga201481gtaatgccac tatactgttt gtgtacatgt ttttgtgtgg acttacattt tcatttctct201541tgggtctgta cctaggagtg gaattgctag gtcatgtggt aactctgtct ttaatatttc201601gaagaactac cacacttttc atagtggctg caccatttta tgttcccgtt ggcaacgtat201661gagggttcca atttctccat tttgtcaaca cttatctttt ttttttatta tagccatcct201721agtgagtgtg aagtgtgatc tcatggtttt gatttgcatt tcttttgtgg ctaacgacat201781tgatcatctt ttcatgtgcc tattggccat ttgtttatct tctttggaga aatgtgtatt201841cacacccttt gcccactttt aattggatta ttaatctttt tacttttgag ttatgaattc201901tttatatatt ctggatacaa ctcccttgtt agatatatga gaaaattgag ccctctttgt201961aaatattttc tctcattctg tgggctgtct tttcactttc tattttttat ttttattttt202021tagagacaaa gtctcactct gtcacccagg ctggggtgca gtagcatgat catggctcat202081tgtagcctcg aattcctggg ctcaagtgat cctttccccc tcagcctcct gagtagctgg202141aactacaggc atgagccacc attcccagcc aatttcattt tattttttat agtgatggag202201tcttactatg ttgcccagtc tggtctcaaa ctcctggcct caagtgatca tcctgcctca202261acctctcaaa gtgctgggat tacaggcctg agccactgca cccagccaat tatcaatatt202321aatcagatct gtacagtggt cacagtccat tctagttcag ttgctctttt ctagaaccta202381tctcccagta atggcacata tggctgtttt tctaggagaa catcagttgc tcttttgctc202441taagaattcg gtgtttcttg ctgtgggatg aaaggttgta gattatccct gtggtaactg202501cactgattta tcatcccaac catggacggt tttgaaaagt aaagagagtg ctaatgtact202561tatgctggac agacagtttt aaatggagac tgtaatgggc gcagtaggat gcgtggtcac202621tctggttatg cgtgttctgt gaatttgttg ttaattggga aatactggta ttaaaacgtt202681ctgtattttt ctcaagagcc acagatatat gctggtccat gcacatacac gagtgcacat202741ttatgcacac acacacatat acttaggcat tcatgcctaa taatacataa ggtagaggta202801ggtagtctgg ctaagttaat gtagaatttt tgctaaaaat aatgttcttg tattattttc202861cttcagactt gttcacaggg gacacctgca ggtcttctaa gtgctattaa ttcttacaca202921aaaagggcat tccagttaat atggtttgtc aggaagaaat acttattaaa atgtcttgga202981ataaatggaa cacaaatgtt tgaaaaggcc catttcacag cttttattca agttataaaa203041tgggaattac ttcaggtgta aataattcat cccaataaat ttgtttcatt tggtgtctat203101cttcctgacc gcctgtgaat atgagatttt atttttggag ccctgtgttg ttaaccactt203161gttagtttaa cacctctcat gaatcagagt tctgaccctc aagggtccag acctctaagt203221cctttttcac tgaagtgttt gtgcttttct ataggcagtc ttccaaacag aagaaggcca203281ttcaaactgc tatccgcaaa aataaagagg caaacgcagt gctggctcgg ctgaacagtg203341agctccagca gcagctcaag gtaaggaatg agtcccagat ttggatgacc aggcttttcc203401ctggctttta aaaagctttt ttttaattta ataggcatat aataattgta catatttata203461gggtggggta catagtgatg tctccatacc tatcatgtac agtgatctga tgtgtccatt203521ggctgaccat ttagttccat ccctgttgat aagacttgcc tcttcgttct caaacccttt203581ttggaaggtg gagaattact gtatttaagt ttcatgggtt ggttcttgca tagtcagagg203641aagactagtg actttaatat gtaggcttca tattgtgtat gagttaaaag gaaagacaat203701ctattacgaa ggctgtttga agcattcttt attatccaaa gcttctgggt aagcccttaa203761ttatttacat cctctctgaa agaggatgta aatacttatt tcagtagaag gaagtcacct203821tggtactatc aaagcccagc ccctcctctg gtgccttgca aatggttcca tttcaccttc203881tcaatgacat cacccctgtt tctcttcatc gttactctcc cctgtgttag tttctcgggg203941ctgctataac aaaatactac aaatgtggtg gcttaagata accaaaaatg attctctcac204001agctctggag gctagaactc tgaaatcaag atattggcag agctatgatc tctttgaagg204061ctctagaaga tgatccttcc ttacctcttc ccagcttctg atgttgctgg cagccctggg204121cattccttgg cttgtagctg cgtcactcca atctctgcct ctgttgtcac atggccttct204181tcctgtatct ctctgtgtct gtgtccaaat ttctcttctt attgtatcct cttataagga204241caccaatcat tggattagga cccaccctaa tctagcatga cctcatctta acttgattac204301atctgcaaag aacctatttg catataaggt cacatttaca gatactgggg gttaggactt204361aaacatatat tttgagagga catgattcta cccactatat cccccagttc tctactggat204421cattcgccat ccacatgtaa atatgctcca gtattgtcca ttacacttgg ccaattactg204481cctcatttct ttggtcccct tcacagcaaa gtttcttagg agtggtttta ttcattatct204541ttactttcaa accagtaatt ttctcttcct tccacttcag ttggcttcgt cctcctccac204601tcacacccgt cttgtggcta aatccaatag ggctgtttta atattattat ttttttttca204661gaagaattct acacaattga tcatttcctc ctctgaaatg gcttttataa ctggtttttg204721acataccata ctctcctggt tttccttctg cttcactggt gcttcccatt ttcattctct204781gctgtctctt ccttccctga cttaactttt ataccccaac cccatttgcc ccaatatttt204841agtatgaaaa ttttcaaaca tacaggaaag ttgaaagaat cataagtgac tgcttgtata204901gacatcactt cagttctaca ataacatttt actatactta tttttaatca catatctacc204961catttatcct tgtgtccatc agttcatctt atttttgaca tattttcaaa taaattgcag205021atatctgtat actttccctg aatattttag catgcgtata attagctaga gttcaatata205081tttttacagt tttttcttat gatgtaaaat gtttatacaa aatgcatgca caaatattat205141tattattatt attattatta ttattattat tattattatt attatttgag atggagtctc205201gctctgtcgc ccacgctgga gtgcggtggc gtgatctctg ctcactgcaa gctccgcctc205261ctgggttcac accattcttc tggcttagcc tcctgagtag ctgggactac aggcgcctgc205321caccacgccc ggccaacttt ttgtattttt agtagaggtg gggtttcact gtgttagcca205381ggttggtctc gatttcctga ccttgtgatc tgcccgccta ggtctcccag agtgctggga205441ttacaggcgt gagccaccgc gccagcgcac gcacaaacct taagtgtaca tttgctgagt205501ttgtaacaag tacatgtact tacggaatcc aaacccccaa aaagatagag aatattacca205561tcaccccagg aagctccctc atgctttttc ctagtcaatt cctgccctac ccacacagag205621gcaatcacta ttctaatttt tataccatag ttttgcctgt tctataactt tatataagcg205681gtgtcttaca acttatactt cttgtaaagc ttctttcact aagtattatg tttttgagat205741tcatttatat tgttgtgtgt atcagtaatt cattcatttt tattgatgag tagtatccca205801ttgggtgaat atgttacagt ttggttatca attctattga cagacacctg agcggtttcc205861agtttttggc aatttggaat aaaactgcat tttttaataa gtctctgtat ggaaatatgt205921tctcatttct cttggattgg gattcgtagg tatcatggta agtgtatgtg attagtttct205981gagaaactgc tagatcattt tccaaggtgg ttgtagcaat ctgtactcct accactaatg206041tatgagagtt ccaattgttc cgcgtctttg ccaacatttg gtgttatgat tcttattaat206101tttagctagc cctgtagtga tatagttttt atcatctcct tgctgactaa ttatgttgtc206161cacttttctt tgtgattatt tacctttcat atcttacttt atgaaatatc ttttcaaatg206221ttatttttct ccaaagtagt ctatagattt attacaaaac caataaaaat cccagcagga206281ttttttgtga gtatagacaa gctaatccca aaacttatat gtaaaggcca aggagctgga206341attgcttaaa caaatttgaa aaggaagaac aaagttggag gaatcacttt actgtcaagc206401tccagtaatc aagatgctga tgcacaggtc agtggaatag gatacagagt cctgaaatag206461accacatgaa aacagccatt tgatctttga caaaggtgca aaatcgattc aatgagaaat206521aatagttttc aacaaattgt tttataacaa tttgtcattt gcatttaaaa aaataacctt206581gatctaagcc tcacatctat acagagatta acttgaaatg agtcatagat gtaaatgtaa206641aacatagagc tataaaattt ttagaagaaa acagaaaaat cttcatgatc tggagttaga206701aataacacca aaagcacaat ctataacaga gaaaaactaa aaaaattgga cttcatcaaa206761attaaaacct tttattttgc caagcacagt taagaaaatt aagagacaag ctgctgagtg206821ggagaaagta tttgcaaaac acgtatttga taaaagactt atatctcaat tatataaaga206881actcttaaaa ctcaataata agaactctca aaaccaaagc atctcaattt aaaaatgggc206941aaaggatttg aacagacact tcactaaaga atatataaag atggcaaata agtacatgag207001aagatattca acatcattag ccatttggga aatgcaaatt taaaccacaa ttacatacca207061ctatacacat actagaattg ctaaaacaaa aaatgtgaca tcaccaagtg ctggcaatta207121gaactctcat ttattggtgg tgggaatgca gaatggtacc accactctga aaaatcgttc207181agcagtttcc tatgaagtta tacatacact aaccatgtga cccagctgtc tctctacctt207241agtgaaatga aaatttatgt tcgctcaaaa atctatatat gaacttacat agcagtttta207301ttcatgatag cccaaacctg gaaacaacac aaatgtcctt caaaagatga atgaataaat207361tgggatacat tcatacaatg gaatagtact caacagagaa aagaaatgat acacaacttg207421gatgaaactc agacattatg cagagtgaga caagccagta tcaaaaggtt gtgtactgta207481tgattccttt tatttgacat tcttgaaaag gtaaaactat agtgatgtga aacaaatcag207541tggctatcag gggttatgca tgagatgaga gtttgtatga gtagtcccag ggaacttttg207601gggttggttg aactgtaatg tgtcttcttt gtggtggtgg ttacacaaat ctgtacacta207661aaagaaaaaa attgttttac cctatgataa ttttaaaaag ttatttttta aatttcaaag207721ttcattctgt tttctaaaac tttgtatgtg aacttgtttt gttttgcatt ttgtttggtt207781ttggttttct cctctctccc tctgctttgg ggtaggccag tttcatccat tgttctctac207841atttggtggg cccttttgga aactgatatt ccccagttct gaagggtttt ttattttcat207901ttttgttgtc gttgattatt tcctcccccc tgccctcacc cccatttttt tctcattctc207961ccttactgat attcagacat tggacttctt agcttagttc tctaattttc tcatattttc208021tcttctgttt tctttctttt ctcttcctac attttcttct ttatttttgt ctatttctgg208081aaacttaatt atatattcta aatcttctgt tgtttaataa ccaataattt tttgttctct208141gtttattttt gacggtatcc tattcttgtt tcattgttgt gatctttttt ttaattatac208201tttaagttct gggaaacatg tgcagaactc gttgtgatct ttttttatgg tcatttttct208261ttcttcttcc tgaatagttt gtgtttgttg tgttgtgttg tatgtgcttt atcttagatg208321ctttcctttg atatctgatg gcccttcttt gctcatattt aaaaggaaga tgaaaacact208381gattagaagt tcattgtacg ttgtaaggct tgtcaactct gagcttcact gtctggcaat208441ttggctcggc tgtatagttg gggaaccccc aaagggagta tgattagctc tttcctcttg208501gtctggttgg agtgtccaga gtggagactg ttttggtcac ctacatggag ggatagagtt208561ggttcagtgg agcaagaggg ctgtaggtct cagcattcca ggatattcat tcactctccc208621tatttctggc ccagtatctt gtcctcaact gtgcttgttt cttgctaatc cagatgctct208681ctgttgtacc ttttcaagaa ataaaccttc agcattctgc cctggtggca ggaatgatct208741ggggatctaa ttgcttctga aacagatttt cagccaacac ttattttagc caactcccat208801ctccccttat gtactcccac tttaataggt atctggtgtt gccaatttga gtctttcagg208861aattctccta cgtaaaatca agttggttct tatttttctt catgttagcc tgggactcag208921ctttctttac ttgatctgta tgagtggaaa agttctaagt ctagtgaaca gaagtctgaa208981gaatcattga aacagaacta tagtcatagc ccctcaatta attcccaggc taagccagtt209041tatagatcca gaacctcttg aatgatgaag gggcatctat gtctccttaa ggaaggaccc209101cactgtgcag ccaaaaattt acactgcaaa tctttctcct agcctttcct aagggggacc209161ctgctgccat ttaccagggt gactgtgcat tagaggaagg gaaataatta gacttttggg209221gaattaccag acactggctc tgaactcaca ctaattccag gagatgcaaa gtgtcactgt209281ggcccatcag agttggggct tagggagatc aggtgatcaa tggagtttta gcttaYgttc209341gtctcacagt ggacccagtg gactcccaaa ctcaccctRt ggttatttcc ctagctctgg209401aatggcatgc ttggaataga catactcagc agctggcaga attcccacat ttattttcct209461ggatttccta agtcatttac cattcaccag tttgccactc agctttcaaa ttttattgct209521atcatcttgt cccttttgtt ttccctgtct tgtgggtccg tgactttgaa aagttgctgt209581attgttttag tggtgctcca ggagggagag agtgtggtac ccctggtcag tcttccatct209641ttacctgaaa cctctatcta tactgttttg ctagatgatg ttatatggtc tcatggcttt209701gaagaccatc cataaactaa tgaaacctaa tttatgtctc aagccctact tctcctttcc209761tctgtgctct agacttgcat atataacttc ttactcaatg ttttctcctt agatatccaa209821tacacatctc agataattaa ttgccaaccc tctccttgaa atactcattt cttatctaat209881cttctcattc tcaatatcac taccagccag ctactttctt catacaaaaa tcccaggagt209941catcctcaga tttttcttct ctcctacatc aactttatca gcaagtccta ccagttctag210001attcaagatt Katctagaat tcatccactt ctcttcatct tcatggcttc taccctgacg210061gaagccattt tcatttctca cctgggtccc tgcagtaacc tcccagttgc tcccctgctt210121ccactctcat cctgctataa tctactgtct acacagtagc cagagtgaaa attttgaatt210181ttaagttggg tcatgttcct tccctgatta gaaacactca atgccaggcc gggtgcagtg210241gcccacgcct gtaatcccag cactttggga ggccgaggtg ggcagatcac gaggtcagga210301gatcgagacc atcctggcta acgtggtgaa accccgtttc tactaaaaat acagaaaatt210361agccgggcgt ggtggcgggY gcctgtagtc ccagctactY gggaggctga ggcaggagaa210421tggcgtgaac ctgggaggcg gagcttgcag tgagccgaga tcgcgccatt gcactccagc210481ctgggcgaca gagtgagact ctgtctcaga aaaaaaaaaa aaaagaaaga aagaaaccct210541caatgccatc cccSctacat ttccatacta tttcctctgc ccacaaggcc ctgtacgatc210601tggctccttg cttaaatctc tgacttcatt tcctgccaat gtcttccttg ttccatctac210661tctggtcaca caggcttttt gttcatcaaa ttcaccaaac tcactcgctc ctcagggcct210721ttgcagttgc ccttttgtct ctgtgaatat ccttcccccc atatcttcat atggctcact210781cctcatttca ttcaagtctt tgctcaagta ttgctaactc tgagaagcct tcccagggca210841ctctctctcc tttctcccat atccccctct gttactctct attatgttat cttatttcct210901tctggaagta tcttgttcat taactttatt atctttctca agtcatttga atataagccc210961tattgctctg atctggcata ttctttgctg ataccatcac agctacagca ttacctcacc211021tgtaggagat agaaaccaag agacttccgg ctgggtggct aggagagcat gtgaataaat211081acttgtgaag ttgtatactg actattagta tgctgattct tctgaatctc acaattcagc211141ctttctacag gattctttcc caccttgact gattctactt gaataccttt atgtgtcccc211201tctcttcctg ccaaaaaaat ccgttttagg cccgaaagtt tgaattccca aagagcagat211261tgdtcagcct dttttctgcc tttcacccag atgttgtacc ttacaccata tgttgtgcag211321aaacttggat gagactttgg tggaggggca actagcgtac ctttttggcc ctcagaccta211381actgagtatg gtgcgggatt acgagccaga gattgcaact agaaatttaa gtcctgttgg211441ctcactgctc agtattggga gtctgttttt gctttctgct ttctcctagt cttttaattt211501ttaatgactc tgtctaccag ctactgatct tgtcaaaaca ggctaaaact agggcagggt211561ggtagagggg agtaggtcag tttcatttca tcacgttcag aaaggagttg agagattaca211621aattgtgaag ccccttttgt ctggtcacca gcattaaatc tgatacctcc cagtcctcct211681gttcttgagt tatacagcac ttgtatttgt gggtccagtt ataagtttcc ttttaaatct211741gatgaagtag ggatgaattc aagaaatgac ccctttgctt ttttgttttt cttgtgtttt211801gtacaactca ggaggttcat caagaacgaa ttgcattgga aaaccaattg gaacaacttc211861gtccggtcac tgtgttgtga cccccccatg gttcaagtga cagtgggtga ccttgtctgc211921caagatcttt cttttgaatg ttttgaaccc aactacttgt catagatgtt tgactgtgtc211981aaaagctgtg agcagcaaaa tataatccat atgacctttt ctcttgcact tcacttgtat212041gttttactgt ggtggaaaaa atacttcaac tgattatcac acttgaaatg ctaattatca212101gtggaatttg tctcctattc ttaagtactt ctgcaaggta ttagatgctg ctccttacca212161aaaatcagtc ttctagcaaa taaaaataac ttagagcatt atttatttaa agaatttatg212221ttttctataa aaccttataa ccaagaacct tcaggatgct tgtaaccaag tactttcagg212281atgctttttg tatgtatatc acatagagta ggttggtttc ctgagtaatt gggattccaa212341ctagatagtc attggtgtga ccataataaa aacaaaagct aaaataaaag catcagattt212401agactggcgc tgtgccctct accactatat gccaaaaatt gcttctctag tgccattttg212461atattatatc tagctataaa taatacatga ctatcgtttt ctattgaatg tcaaagactt212521actgggtctt tacacctctg taaagtggag gttttggcaa tgagctgttt aacttggcca212581agctaggctg tcaacccaga caactcagtc ctttcatgta aatttttggg caagagaagt212641ctgctttgac tgccttgcct gaccaaatta aataaatagt tgccagatct ctacttttat212701cctgcatggg ataatatatc tgtaaatgca tgagtttgga aaccacccat caaatatgaa212761aggctgatga gactgttagg attgataaac atttgttgtt tgaacccagc tatccaaact212821ggggaagaag gggaaatggt tgttaccaac aatctctagt agttatttta atctttattt212881taacctggat tgcaaatgta tggggtatga ggagataatg aaagaaaagt ggtcctcatg212941ctggatatgt gactgttttg ttctctgtaa agtgtattct tgggaagtga ttggtttata213001tcagattcag aggagcccaa ttaactccag tgtaggtaat gtaaaacagt aagctcaaat213061tgcagaagcc Rgtatcctac agaaatttaa gtagctactg cttctcctga aagcccaagt213121tgaaaatggg accacagggg cacagggcta ctgaccctca cagggcatgc cagcgttaac213181accactttga gagaccaatg gcaagaaatg ctgtctcttg tgcattttac taatttcccc213241attcttaggg tagcagggtg ggggtgtata gggtcttttt gaagcagttt tggataggtt


Following are NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO and REPS2 complementary DNA sequences (cDNAs; SEQ ID NO: 8, 9, 10, 11, 12, 13, 14, 15, 16 and 17, respectively). Following is a NRP1 cDNA sequence (SEQ ID NO: 8).

>gi|4505456|ref|NM_003873.1− Homo sapiens neuropilin 1 (NRP1), mRNAATGGAGAGGGGGCTGCCGCTCCTCTGCGCCGTGCTCGCCCTCGTCCTCGCCGCGGCCGGCGCTTTTCGCAACCATGAATGTGGCGATACTATAAAAATTGAAAGCCCCGGGTACCTTACATCTCCTGGTTATCCTCATTCTTATCACCCAAGTGAAAAATGCGAATGGCTGATTCAGGCTCCGGACCCATACCAGAGAATTATGATCAACTTCAACCCTCACTTCGATTTGGAGGACACAGACTGCAAGTATGACTACGTGGAAGTCTTCGATGGAGAAAATGAAAATGGACATTTTAGGGGAAAGTTCTGTGGAAAGATAGCCCCTCCTCCTGTTGTGTCTTCAGGGCCATTTCTTTTTATCAAATTTGTCTCTGACTACGAAACACATGGTGCAGGATTTTCCATACGTTATGAAATTTTCAAGAGAGGTCCTGAATGTTCCCAGAACTACACAACACCTAGTGGAGTGATAAAGTCCCCCGGATTCCCTGAAAAATATCCCAACAGCCTTGAATGCACTTATATTGTCTTTGCGCCAAAGATGTCAGAGATTATCCTGGAATTTGAAAGCTTTGACCTGGAGCCTGACTCAAATCCTCCAGGGGGGATGTTCTGTCGCTACGACCGGCTAGAAATCTGGGATGGATTCCCTGATGTTGGCCCTCACATTGGGCGTTACTGTGGACAGAAAACACCAGGTCGAATCCGATCCTCATCGGGCATTCTCTCCATGGTTTTTTACACCGACAGCGCGATAGCAAAAGAAGGTTTCTCAGCAAACTACAGTGTCTTGCAGAGCAGTGTCTCAGAAGATTTCAAATGTATGGAAGCTCTGGGCATGGAATCAGGAGAAATTCATTCTGACCAGATCACAGCTTCTTCCCAGTATAGCACCAACTGGTCTGCAGAGCGCTCCCGCCTGAACTACCCTGAGAATGGGTGGACTCCCGGAGAGGATTCCTACCGAGAGTGGATACAGGTAGACTTGGGCCTTCTGCGCTTTGTCACGGCTGTCGGGACACAGGGCGCCATTTCAAAAGAAACCAAGAAGAAATATTATGTCAAGACTTACAAGATCGACGTTAGCTCCAACGGGGAAGACTGGATCACCATAAAAGAAGGAAACAAACCTGTTCTCTTTCAGGGAAACACCAACCCCACAGATGTTGTGGTTGCAGTATTCCCCAAACCACTGATAACTCGATTTGTCCGAATCAAGCCTGCAACTTGGGAAACTGGCATATCTATGAGATTTGAAGTATACGGTTGCAAGATAACAGATTATCCTTGCTCTGGAATGTTGGGTATGGTGTCTGGACTTATTTCTGACTCCCAGATCACATCATCCAACCAAGGAGACAGAAACTGGATGCCTGAAAACATCCGCCTGGTAACCAGTCGCTCTGGCTGGGCACTTCCACCCGCACCTCATTCCTACATCAATGAGTGGCTCCAAATAGACCTGGGGGAGGAGAAGATCGTGAGGGGCATCATcATTCAGGGTGGGAAGCACCGAGAGAACAAGGTGTTCATGAGGAAGTTCAAGATCGGGTACAGCAACAACGGCTCGGACTGGAAGATGATCATGGATGACAGCAAACGCAAGGCGAAGTCTTTTGAGGGCAACAACAACTATGATACACCTGAGCTGCGGACTTTTCCAGCTCTCTCCACGCGATTCATCAGGATCTACCCCGAGAGAGCCACTCATGGCGGACTGGGGCTCAGAATGGAGCTGCTGGGCTGTGAAGTGGAAGCCCCTACAGCTGGACCGACCACTCCCAACGGGAACTTGGTGGATGAATGTGATGACGACCAGGCCAACTGCCACAGTGGAACAGGTGATGACTTCCAGCTCACAGGTGGCACCACTGTGCTGGCCACAGAAAAGCCCACGGTCATAGACAGCACCATACAATCAGAGTTTCCAACATATGGTTTTAACTGTGAATTTGGCTGGGGCTCTCACAAGACCTTCTGCCACTGGGAACATGACAATCACGTGCAGCTCAAGTGGAGTGTGTTGACCAGCAAGACGGGACCCATTCAGGATCACACAGGAGATGGCAACTTCATCTATTCCCAAGCTGACGAAAATCAGAAGGGCAAAGTGGCTCGCCTGGTGAGCCCTGTGGTTTATTCCCAGAACTCTGCCCACTGCATGACCTTCTGGTATCACATGTCTGGGTCCCACGTCGGCACACTCAGGGTCAAACTGCGCTACCAGAAGCCAGAGGAGTACGATCAGCTGGTCTGGATGGCCATTGGACACCAAGGTGACCACTGGAAGGAAGGGCGTGTCTTGCTCCACAAGTCTCTGAAACTTTATCAGGTGATTTTCGAGGGCGAAATCGGAAAAGGAAACCTTGGTGGGATTGCTGTGGATGACATTAGTATTAATAACCACATTTCACAAGAAGATTGTGCAAAACCAGCAGACCTGGATAAAAAGAACCCAGAAATTAAAATTGATGAAACAGGGAGCACGCCAGGATACGAAGGTGAAGGAGAAGGTGACAAGAACATCTCCAGGAAGCCAGGCAATGTGTTGAAGACCTTAGAACCCATCCTCATCACCATCATAGCCATGAGCGCCCTGGGGGTCCTCCTGGGGGCTGTCTGTGGGGTCGTGCTGTACTGTGCCTGTTGGCATAATGGGATGTCAGAAAGAAACTTGTCTGCCCTGGAGAACTATAACTTTGAACTTGTGGATGGTGTGAAGTTGAAAAAAGACAAACTGAATACACAGAGTACTTATTCGGAGGCATGA


Following is a NID2 cDNA sequence (SEQ ID NO: 9).

>gi|33469988|ref|NM_007361.2| Homo sapiens nidogen 2 (osteonidogen) (NID2), mRNAGCCTTAGAAAAGTTAACGAGAACCAGATGTGGTGGCCACTGCCGAACTTTCTCAGAGCCGGTGATTGGTCCCCAGCCGAGGGCCTCAGCCAATTAGCTTGCTGGGTGGGCCTGGAGTCCCGCCCCGCCCAGGCGCCCGCGGAGATCCAGGTTCGAGGCTGGCGGGGCGCGGAGAGTGGGCTGGAGGCCGGGGCGGGACGCGTTGTGCAGCGGGTAAGCGCACGGCCGAGCGAGCATGGAGGGGGACCGGGTGGCCGGGCGGCCGGTGCTGTCGTCGTTACCAGTGCTACTGCTGCTGCAGTTGCTAATGTTGCGGGCCGCGGCGCTGCACCCAGACGAGCTCTTCCCACACGGGGAGTCGTGGGGGGACCAGCTCCTCCAGGAAGGCGACGACGAAAGCTCAGCCGTGGTGAAGCTGGCGAATCCCCTGCACTTCTACGAAGCCCGATTCAGCAACCTCTACGTGGGCACCAACGGCATCATCTCCACTCAGGACTTCCCCAGGGAAACGCAGTATGTGGACTATGATTTCCCCACCGACTTCCCGGCCATCGCCCCTTTTCTGGCGGACATCGACACGAGCCACGGCAGAGGCCGAGTCCTGTACCGAGAGGACACCTCCCCCGCAGTGCTGGGCCTGGCCGCCCGCTATCTGCGCGCTGGCTTCCCGCGCTCTGCGCGCTTTACCCCCACCCACGCCTTCCTGGCCACCTGGGAGCAGGTAGGCCCTTACGAGGAGGTCAAGCGCGGGGCGCTGCCCTCGGGAGAGCTGAACACTTTCCAGGCAGTTTTGGCATCTGATGGGTCTGATAGCTACGCCCTCTTTCTTTATCCTGCCAACGGCCTGCAGTTCCTTGGAACCCGCCCCAAAGAGTCTTACAATGTCCAGCTTCAGCTTCCAGCTCGGGTGGGCTTCTGCCGAGGGGAGGCTGATGATCTGAAGTCAGAACGACCATATTTCAGCTTGACTAGCACTGAACAGTCTCTGAAAAATCTCTATCAACTAAGCAACCTGGGGATCCCTGGAGTCTGGGCTTTCCATATCGGCAGCACTTCCCCGTTGGACAATGTCAGGCCAGCTGCAGTTGGAGACCTTTCCGCTGCCCACTCTTCTGTTCCCCTGGGACGTTCCTTCAGCCATGCTACAGCCCTGGAAAGTGACTATAATGAGGACAATTTGGATTACTACGATGTGAATGAGGAGGAAGCTGAATACCTTCCGGGTGAACCAGAGGAGGCATTGAATCGCCACAGCAGCATTGATGTTTCCTTCCAATCCAAAGTGGATACAAAGCCTTTAGAGGAATCTTCCACCTTGGATCCTCACACCAAAGAAGGAACATCTCTGGGAGAGGTAGGGGGCCCAGATTTAAAAGGCCAAGTTGAGCCCTGGGATGAGAGAGAGACCAGAAGCCCAGCTCCACCAGAGGTAGACAGAGATTCACTGGCTCCTTCCTGGGAAACCCCACCACCGTACCCCGAAAACGGAAGCATCCAGCCCTACCCAGATGGAGGGCCAGTGCCTTCGGAAATGGATGTTCCCCCAGCTCATCCTGAAGAAGAAATTGTTCTTCGAAGTTACCCTGCTTCAGGTCACACTACACCCTTAAGTCGAGGGACGTATGAGGTGGGACTGGAAGACAACATAGGTTCCAACACCGAGGTCTTCACGTATAATGCTGCCAACAAGGAAACCTGTGAACACAACCACAGACAATGCTCCCGGCATCCCTTCTGCACGGACTATGCCACTGGCTTCTGCTGCCACTGCCAATCCAAGTTTTATGGAAATCGGAAGCACTGTCTGCCTGAGGGGGCACCTCACCGAGTGAATGGGAAAGTGAGTGGCCACCTCCACGTGGGCCATACACCCGTGCACTTCACTGATGTGGACCTGCATGCGTATATCGTGGGCAATGATGGCAGAGCCTACACGGCCATCAGCCACATCCCACAGCCAGCAGCCGAGGCCCTCCTCCCCCTCACACCAATTGGAGGCCTGTTTGGCTGGCTCTTTGCTTTAGAAAAACCTGGCTCTGAGAACGGCTTCAGCCTCGCAGGTGCTGCCTTTACCCATGACATGGAAGTTACATTCTACCCGGGAGAGGAGACGGTTCGTATCACTCAAACTGCTGAGGGACTTGACCCAGAGAACTACCTGAGCATTAAGACCAACATTCAAGGCCAGGTGCCTTACGTCCCAGCAAATTTCACAGCCCACATCTCTCCCTACAAGGAGCTGTACCACTACTCCGACTCCACTGTGACCTCTACAAGTTCCAGAGACTACTCTCTGACTTTTGGTGCAATCAACCAAACATGGTCCTACCGCATCCACCAGAACATCACTTACCAGCTGTGCAGGCACGCCCCCAGACACCCGTCCTTCCCCACCACCCAGCAGCTGAACGTGGACCGGGTCTTTGCCTTGTATAATGACGAAGAAAGAGTGCTTAGATTTGCTGTGACCAATCAAATTGGCCCGGTCAAAGAAGATTCAGACCCCACTCCGGGGAATCCTTGCTATGATGGGAGCCACATGTGTGACACAACAGCACGGTGCCATCCAGGGACAGGTGTAGATTACACCTGTGAGTGCGCATCTGGGTACCAGGGAGATGGACGGAACTGTGTGGATGAAAATGAATGTGCAACTGGCTTTCATCGCTGTGGCCCCAACTCTGTATGTATCAACTTGCCTGGAAGCTACAGGTGTGAGTGCCGGAGTGGTTATGAGTTTGCAGATGACCGGCATACTTGCATCTTGATCACCCCACCTGCCAACCCCTGTGAGGATGGCAGTCATACCTGTGCTCCTGCTGGGCAGGCCCGGTGTGTTCACCATGGAGGCAGCACGTTCAGCTGTGCCTGCCTGCCTGGTTATGCCGGCGATGGGCACCAGTGCACTGATGTAGATGAATGCTCAGAAAACAGATGTCACCCTGCAGCTACCTGCTACAATACTCCTGGTTCCTTCTCCTGCCGTTGTCAACCCGGATATTATGGGGATGGATTTCAGTGCATACCTGACTCCACCTCAAGCCTGACACCCTGTGAACAACAGCAGCGCCATGCCCAGGCCCAGTATGCCTACCCTGGGGCCCGGTTCCACATCCCCCAATGCGACGAGCAGGGCAACTTCCTGCCCCTACAGTGTCATGGCAGCACTGGTTTCTGCTGGTGCGTGGACCCTGATGGTCATGAAGTTCCTGGTACCCAGACTCCACCTGGCTCCACCCCGCCTCACTGTGGACCATCACCAGAGCCCACCCAGAGGCCCCCGACCATCTGTGAGCGCTGGAGGGAAAACCTGCTGGAGCACTACGGTGGCACCCCCCGGGATGACCAGTACGTGCCCCAGTGCGATGACCTGGGCCACTTCATCCCCCTGCAGTGCCACGGAAAGAGCGACTTCTGCTGGTGTGTGGACAAAGATGGCAGAGAGGTGCAGGGCACCCGCTCCCAGCCAGGCACCACCCCTGCGTGTATACCCACCGTCGCTCCACCCATGGTCCGGCCCACGCCCCGGCCAGATGTGACCCCTCCATCTGTGGGCACCTTCCTGCTCTATACTCAGGGCCAGCAGATTGGCTACTTACCCCTCAATGGCACCAGGCTTCAGAAGGATGCAGCTAAGACCCTGCTGTCTCTGCATGGCTCCATAATCGTGGGAATTGATTACGACTGCCGGGAGAGGATGGTGTACTGGACAGATGTTGCTGGACGGACAATCAGCCGTGCTGGTCTGGAACTGGGAGCAGAGCCTGAGACGATCGTGAATTCAGGTCTGATAAGCCCTGAAGGACTTGCCATAGACCACATCCGCAGAACAATGTACTGGACGGACAGTGTCCTGGATAAGATAGAGAGCGCCCTGCTGGATGGCTCTGAGCGCAAGGTCCTCTTCTACACAGATCTGGTGAATCCCCGTGCCATCGCTGTGGATCCAATCCGAGGCAACTTGTACTGGACAGACTGGAATAGAGAAGCTCCTAAAATTGAAACGTCATCTTTAGATGGAGAAAACAGAAGAATTCTGATCAATACAGACATTGGATTGCCCAATGGCTTAACCTTTGACCCTTTCTCTAAACTGCTCTGCTGGGCAGATGCAGGAACCAAAAAACTGGAGTGTACACTACCTGATGGAACTGGACGGCGTGTCATTCAAAACAACCTCAAGTACCCCTTCAGCATCGTAAGCTATGCAGATCACTTCTACCACACAGACTGGAGGAGGGATGGTGTTGTATCAGTAAATAAACATAGTGGCCAGTTTACTGATGAGTATCTCCCAGAACAACGATCTCACCTCTACGGGATAACTGCAGTCTACCCCTACTGCCCAACAGGAAGAAAGTAAGTACAGTAATGTAAAGGAAGACTTGGAGTTTACAATCAGAACCTGGACCCTAAAGAACAGTGACTGCAAAGGCAAAGAAAGTAAAAAAGGAATTGGCCATTAGACGTTCCTGAGCATCCAAGATGAACATTTTGTAGTGCAAAAAGACTTTTGTGAAAAGCTGATACCTCAATCTTTACTACTGTATTTTTAAAAATGAAGGTTGTTATTGCAAGTTTAAAAAGGTAACAGAATTTTAACTGTTGCTTATTAAAGCAACTTCTTGTAAACATTTATCATTAATATTTAAAAGATCAAATTCATTCAACTAAGAATTAGAGTTTAAGACTCTAAACCTGATTTTTGCCATGGATTCCTTCTGGCCAAGAAATTAAAGCACATGTGATCAATATAACAATATAATCCTAAACCTTGACAGTTGGAGAAGCCAATGCAGAACTGATGGGAAAGGACCAATTATTTATAGTTTCCCAACAAAAGTTCTAAGATTTTTTACCTCTGCATCAGTGCATTTCTATTTATATCAAAAGGTGCTAAAATGATTCAATTTGCATTTTCTGATCCTGTAGTGCCTCTATAGAAGTACCCACAGAAAGTAAAGTATCACATTTATAAATACCAAAGATGTAACAATTTTAAAATTTTCTAGATTACTCCAATAAAGTGTTTTAAGTTTTCCTATGAAAAAAAAAAAAAAAAAAAA


Following is a ENDO180 cDNA sequence (SEQ ID NO: 10).

>gi|5174484|ref|NM_006039.1| Homo sapiens mannose receptor, C type 2 (MRC2), mRNACGGAGGAGGACGCGAGCCCCTTGCGGGCGGTCATCACAGCCCAGCCTCGGGGCTGCCACAGCGCGTTGCGCCTGTGCGCCCTCGGTCCCCGCGTCCACTGAGCGCCGCGCTCGGGGATGGGGCCCGGCCGGCCGGCCCCCGCGCCCTGGCCTCGTCACCTGCTGCGCTGCGTCCTGCTCCTCGGGTGCCTGCACCTCGGCCGTCCCGGCGCCCCTGGGGACGCCGCCCTCCCGGAACCCAACGTCTTCCTCATCTTCAGCCATGGACTGCAGGGCTGCCTGGAGGCCCAGGGCGGGCAGGTCAGAGTCACCCCGGCTTGCAATACCAGCCTCCCTGCCCAGCGCTGGAAGTGGGTCTCCCGAAACCGGCTATTCAACCTGGGTACCATGCAGTGCCTGGGCACAGCCTGGCCAGGCACCAACACCACGGCCTCCCTGGGCATCTATGAGTGTGACCGGGAAGCACTGAATCTTCGCTGGCATTGTCGTACACTGGGTGACCAGCTGTCCTTGCTCCTGGGGGCCCGCACCAGCAACATATCCAACCCTGGCACCCTTGAGCGTGGTGACCAGACCCGCAGTGGCCAGTGGCGCATCTACGGCAGCGAGGAGGACCTATGTGCTCTGCCCTACCACGAGGTCTACACCATCCAGGGAAACTCCCACGGAAAGCCGTCCACCATCCCCTTCAAATATGACAACCAGTGGTTCCACGGCTGCACCAGCACGGGCCGCGAGGATGGTCACCTGTGGTGTCCCACCACCCAGGACTACGGCAAAGACGAGCGCTGGGGCTTCTGCCCCATCAAGAGTAACGACTGCGAGACCTTCTGGGACAAGGACCAGCTGACTGACAGCTGCTACCAGTTTAACTTCCAGTCCACGCTGTCGTGGAGGGAGCCCTGGGCCAGCTGCGAGCAGCAGGGTGCCGATCTCCTGAGCATCACGGAGATCCACGAGCACACCTACATCAACGGCCTCCTCACTCGGTACAGCTCCACCCTGTGGATCGGCTTGAATCACTTGGACACGACCGGAGGCTGGCAGTGGTCGGACAACTCGCCCCTCAAGTACCTCAACTGGGAGAGTGACCAGCCGGACAACCCCAGTGAGGAGAACTGTGGAGTGATCCGCACTGAGTCCTCGGGCGGCTGGCAGAACCGTGACTGCAGCATCGCGCTGCCCTATGTGTGCAAGAAGAAGCCCAACGCCACGGCCGAGCCCACCCCTCCAGACAGGTGGGCCAATGTGAAGGTGGAGTGCGAGCCGAGCTCCCAGCCCTTCCAGGGCCACTGCTACCGCCTGCAGGCCGACAAGCGCAGCTGGCAGGAGTCCAAGAAGGCATGTCTACGGGGCGGTGGCGACCTGGTCAGCATCCACAGCATGGCGGAGCTGGAATTCATCACCAAGCAGATCAAGCAAGAGGTCGAGGAGCTGTGGATCGGCCTCAACGATTTGAAACTGCAGATGAATTTTGAGTGGTCTGACGGGAGCCTTGTGACCTTCACCCACTGGCACCCCTTTGAGCCCAACAACTTCCGGGACAGTCTGGAGGACTGTGTCACCATCTGGGGCCCGGAAGGCCGCTGGAACGACAGTCCCTGTAACCAGTCCTTGCCATCCATCTGCAAGAAGGCAGGCCAGCTGAGCCAGGGGGCCGCCGAGGAGGACCATGGCTGCCGGAAGGGTTGGACGTGGCACAGCCCATCCTGCTACTGGCTGGGAGAAGACCAAGTGACCTACAGTGAGGCCCGGCGCCTGTGCACTGACCATGGCTCTCAGCTGGTCACCATCACCAACAGGTTCGAGCAGGCCTTCGTCAGCAGCCTCATCTACAACTGGGAGGGCGAGTACTTCTGGACGGCCCTGCAGGACCTCAACAGCACCGGCTCCTTCTTCTGGCTCAGTGGGGATGAAGTCATGTACACCCACTGGAACCGGGACCAGCCCGGGTACAGCCGTGGGGGCTGCGTGGCGCTGGCCACTGGCAGCGCCATGGGGCTGTGGGAGGTGAAGAACTGTACCTCGTTCCGGGCCCGCTACATCTGCCGGCAGAGCCTGGGCACTCCAGTGACGCCGGAGCTGCCCGGGCCAGATCCCACGCCCAGCCTCACTGGCTCCTGTCCCCAGGGCTGGGCCTCGGACACCAAACTCCGGTATTGCTATAAGGTGTTCAGCTCAGAGCGGCTGCAGGACAAGAAGAGCTGGGTCCAGGCCCAGGGGGCCTGCCAGGAGCTGGGGGCCCAGCTGCTGAGCCTCCCCAGCTACGAGGAGGAGCACTTTGTGGCCAACATGCTCAACAAGATCTTCGGTGAATCAGAACCCGAGATCCACGAGCAGCACTGGTTCTGGATCGGCCTGAACCGTCGGGATCCCAGAGGGGGTCAGAGTTGGCGCTGGAGCGACGGCGTAGGCTTCTCTTACCACAATTTCGACCGGAGCCGGCACCACGACGACGACATCCGAGGCTGTGCGGTGCTGGACCTGGCCTCCCTGCACTGGGTGGCCATGCAGTGCGACACACAGCTGGACTGGATCTGCAAGATCCCCAGAGGTACGGACGTGCGGGAGCCCGACGACAGCCCTCAAGGCCGACGGGAATGGCTGCGCTTCCAGGAGGCCGAGTACAAGTTCTTTGAGCACCACTCCACGTGGGCGCAGGCGCAGCGCATCTGCACGTGGTTCCAGGCCGAGCTGACCTCCGTGCACAGCCAGGCAGAGCTAGACTTCCTGAGCCACAACTTGCAGAAGTTCTCCCGGGCCCAGGAGCAGCACTGGTGGATCGGCCTGCACACCTCTGAGAGCGATGGGCGCTTCAGATGGACAGATGGTTCCATTATAAACTTCATCTCCTGGGCACCAGGCAAACCTCGGCCTGTCGGCAAGGACAAGAAGTGCGTGTACATGACAGCCAGCCGAGAGGACTGGGGGGACCAGAGGTGCCTGACAGCCTTGCCCTACATCTGCAAGCGCAGCAACGTCACCAAAGAAACGCAGCCCCCAGACCTGCCAACTACAGCCCTGGGGGGCTGCCCCTCTGACTGGATCCAGTTCCTCAACAAGTGTTTTCAGGTCCAGGGCCAGGAACCCCAGAGCCGGGTGAAGTGGTCAGAGGCACAGTTCTCCTGTGAACAGCAAGAGGCCCAGCTGGTCACCATCACAAACCCCTTAGAGCAAGCATTCATCACAGCCAGCCTGCCCAATGTGACCTTTGACCTTTGGATTGGCCTCCATGCCTCGCAGAGGGACTTCCAGTGGGTGGAGCAGGAGCCTTTGATGTATGCCAACTGGGCACCTGGGGAGCCCTCTGGCCCTAGCCCTGCTCCCAGTGGCAACAAACCGACCAGCTGTGCGGTGGTCCTGCACAGCCCCTCAGCCCACTTCACTGGCCGCTGGGACGATCGGAGCTGCACGGAGGAGACCCATGGCTTCATCTGCCAGAAGGGCACGGACCCCTCCCTGAGCCCGTCCCCAGCAGCGCTGCCCCCCGCCCCGGGCACTGAGCTCTCCTACCTCAACGGCACCTTCCGGCTGCTTCAGAAGCCGCTGCGCTGGCACGATGCCCTCCTGCTGTGTGAGAGCCACAATGCCAGCCTGGCCTACGTGCCCGACCCCTACACCCAGGCCTTCCTCACGCAGGCTGCCCGAGGGCTGCGCACGCCGCTCTGGATTGGGCTGGCTGGCGAGGAGGGCTCTCGGCGGTACTCCTGGGTCTCAGAGGAGCCGCTGAACTACGTGGGCTGGCAGGACGGGGAGCCGCAGCAGCCGGGGGGCTGTACCTACGTAGATGTGGACGGGGCCTGGCGCACCACCAGCTGTGACACCAAGCTGCAGGGGGCTGTGTGTGGGGTTAGCAGTGGGCCCCCTCCTCCCCGAAGAATAAGCTACCATGGCAGCTGTCCCCAGGGACTGGCAGACTCCGCGTGGATTCCCTTCCGGGAGCACTGCTATTCTTTCCACATGGAGCTGCTGCTGGGCCACAAGGAGGCGCGACAGCGCTGCCAGAGAGCGGGTGGGGCCGTCCTGTCTATCCTGGATGAGATGGAGAATGTGTTTGTCTGGGAGCACCTGCAGAGCTATGAGGGCCAGAGTCGGGGCGCCTGGCTGGGCATGAACTTCAACCCCAAAGGAGGCACTCTGGTCTGGCAGGACAACACAGCTGTGAACTACTCCAACTGGGGGCCCCCGGGCTTGGGCCCCAGCATGCTGAGCCACAACAGCTGCTACTGGATTCAGAGCAACAGCGGGCTATGGCGCCCCGGCGCTTGCACCAACATCACCATGGGTGTCGTCTGCAAGCTTCCTCGTGCTGAGCAGAGCAGCTTCTCCCCATCAGCGCTTCCAGAGAACCCAGCGGCCCTGGTGGTGGTGCTGATGGCGGTGCTGCTGCTCCTGGCCTTGCTGACCGCAGCCCTCATCCTTTACCGGAGGCGCCAGAGCATCGAGCGCGGGGCCTTTGAGGGTGCCCGCTACAGCCGCAGCAGCTCCAGCCCCACCGAGGCCACTGAGAAGAACATCCTGGTGTCAGACATGGAAATGAATGAGCAACAAGAATAGAGCCAGGCGCGTGGGCAGGGCCAGGGCGGGAGGAGCTGGGGAGCTGGGGCCCTGGGTCAGTCTGGCCCCCCACCAGCTGCCTGTCCAGTTGGCCTATGGAAGGGTGCCCTTGGGAGTCGCTGTTGGGAGCCGGAGCTGGGCAGAGCCTGGGCTGGTGGGGTGCCACCCTCCCACAAGGGCTGGGCTGAGACCCAGCTGAGTGCAGCGTGGCGTTTCCCTTTCTGGGGGGGCCTGAGGTCTTGTCACCTGGTCCTGTGCCCCCACAGGAACCAGAGGTAGGATGGGAGGGGGAACGAGAGCCTCTTTCTCCCCAGAGCCCCCGGCCCAGGCCTGTTGATCCGCGCCCCAGGACCCCCTTCTTTGCAGAGCCCGAGGAGCCTCCCCTGTCCCCTCGGGCAGATCTGTTGTGTCTCTCTTCCCACCTGGCAGCCTCAGCTCTGTGCCCCTCACCCTGCTCCCTCTCGCCCCTTCTCTCCCACCCCTTCCTTCTGAGCCGGGCCCTGGGGATTGGGGAGCCCTCTTGTTCCTGATGAGGGTCAGCTGAGGGGGCTGAGCATCCATCACTCCTGTGCCTGCTGGGGTGGCTGTGGGGCGTGGCAGGAGGGGCCTAGGTGGGTTGGGCCTGAGAACCAGGGCACGGGTGTGGTGTCTGCTGGGCTGGAGATAAGACTGGGGAGAGACACCCCAACCTCCCAGGGTGGGAGCTGGGCCGGGCTGGGATGTCATCTCCTGCCGGGCGGGGGAGGGCTCTGCCCCTGGAAGAGTCCCCTGTGGGGACCAAAATAAGTTCCCTAACATCTCCAGCTCCTGGCTCTGGTTTGGAGCAAGGGGAAGGGTTGCCAGAGTCCTGGGGGCCCCAGAGGAGCAGGAGTCTGGGAGGGCCCAGAGTTCACCCTCTAGTGGATCCAGGAGGAGCAGCACCCGAGCCCTGGAGTGGCCCAGTACCCTTCCAAGAGGCCACAGTCCCAGCCAGGACAAAGTATGCGGCCCATCCTGGTGCGACAGCGTGGGACAATGTGAACATGGACTCGAAGACATGGCCCTTTCTCTGTAGTTGATTTTTTAAATGTGCCATTATTGTTTTT


Following is a first CDK10 cDNA sequence (SEQ ID NO: 11).

>gi|32528262|ref|NM_003674.2| Homo sapiens cyclin-dependent kinase (CDC2-like) 10(CDK10), transcript variant 1, mRNAGTCTGCGCCTGCGCGCAAGAGAGGCGGGGCCAGCGCTCGGCATGGCGGAGCCAGATCTGGAGTGCGAGCAGATCCGTCTGAAGTGTATTCGTAAGGAGGGCTTCTTCACGGTGCCTCCGGAACACAGGGTGACCCACCGCCCCCAGCCTGGCCTGGCATTTCTTTGAGTTCAGGAAGTGTGACAAGGATTTGGACACCCAGAAATAAGCGTGTCGAGAAGAGCACAAGCAGAGGATCTGGGACCATGCCGGAGTGTGAAGGAGTTTGAGAAGCTGAACCGCATTGGAGAGGGTACCTACGGCATTGTGTATCGCGCCCGGGACACCCAGACAGATGAGATTGTCGCACTGAAGAAGGTGCGGATGGACAAGGAGAAGGATGGCATCCCCATCAGCAGCTTGCGGGAGATCACGCTGCTGCTCCGCCTGCGTCATCCGAACATCGTGGAGCTGAAGGAGGTGGTTGTGGGGAACCACCTGGAGAGCATCTTCCTGGTGATGGGTTACTGTGAGCAGGACCTGGCCAGCCTCCTCGAGAATATGCCAACACCCTTCTCGGAGGCTCAGGTCAAGTGCATCGTGCTGCAGGTGCTCCGGGGCCTCCAGTATCTGCACAGGAACTTCATTATCCACAGGGACCTGAAGGTTTCCAACTTGCTCATGACCGACAAGGGTTGTGTGAAGACAGCGGATTTCGGCCTGGCCCCGGCCTATGGTGTCCCAGTAAAGCCAATGACCCCCAAGGTGGTCACTCTCTCGTACCGAGCCCCTGAACTGCTGTTGGGAACCACCACGCAGACCACCAGCATCGACATGTGGGCTGTGGGCTGCATACTCGCCGAGCTGCTGGCGCACAGGCCTCTTCTCCCCGGCACTTCCGAGATCCACCAGATCGACTTGATCGTGCAGCTGCTGGGCACGCCCAGTGAGAACATCTGGCCGGGCTTTTCCAAGCTGCCACTGGTCGGCCAGTACAGCCTCCGGAAGCAGCCCTACAACAACCTGAAGCACAAGTTCCCATGGCTGTCGGAGGCCGGGCTGCGCCTGCTGCACTTCCTGTTCATGTACGACCCTAAGAAAAGGGCGACGGCCGGGGACTGCCTGCAGAGCTCCTATTTCAAGGAGAAGCCCCTACCCTGTGAGCCGGAGCTCATGCCGACCTTTCCCCACCACCGCAACAAGCGGGCCGCCCCAGCCACCTCCGAGGGCCAGAGCAAGCGCTGTAAACCCTGACGGTGGGCCTGGCACACGCCTGTATTCCCACACCAGGTCTTCCGATCAGTGGTGTCTGTGAAGGGTGCCGCGAGCCAGGCTGACCAGGCGCCCGGGATCCAGCTCATCCCCTTGGCTGGGAACATCCTCCACTGACTTCCTCCCACTGTCTGCCCTGAACCCACTGCTGCCCCCAGAAAAAGGCCGGGTGACACCGGGGGGCTCCCAGCCCCTGCACCCTGGAAGGGCAGGTCTGGCGGCTCCATCCGTGGCTGCAGGGGTCTCATGTGGTCCTCCTCGCTATGTTGGAAATGTGCAACCACTGCTTCTTGGGAGGAGTGGTGGGTGCAGTCCCCCCGCTGTCTTTGAGTTGTGGTGGACGCTGGCCTGGGATGAGAGGGCCCAGAAGACCTTCGTATCCCCTCTCAGTCGCCCGGGGCTGTCCCGTGCATGGGTTGGCTGTGGGGACCCCAGGTGGGCCTGGCAGGACTCCAGATGAGGACAAGAGGGACAAGGTATGGGGTGGGAGCCACAATTGAGGATACCCCGAGACTACCAGGAGAGCCCTGGGCTGGAGGCTGAGCTGCATCCCTGCTCCCCACATGGAGGACCCAACAGGAGGCCGTGGCTCTGATGCTGAGCGAAGCTATAGGCTCTTGTTGGATAAAAGCTTTTTTAACA


Following is a seco-nd CDK10 cDNA sequence (SEQ ID NO: 12).

>gi|32528264|ref|NM_052987.2| Homo sapiens cyclin-dependent kinase (CDC2-like) 10(CDK10), transcript variant 2, mRNAGTCTGCGCCTGCGCGCAAGAGAGGCGGGGCCAGCGCTCGGCATGGCGGAGCCAGATCTGGAGTGCGAGCAGATCCGTCTGAAGTGTATTCCTGGGACGATGCCGGAGTGTGAAGGAGTTTGAGAAGCTGAACCGCATTGGAGAGGGTACCTACGGCATTGTGTATCGGGCCCGGGACACCCAGACAGATGAGATTGTCGCACTGAAGAAGGTGCGGATGGACAAGGAGAAGGATGGCATCCCCATCAGCAGCTTGCGGGAGATCACGCTGCTGCTCCGCCTGCGTCATCCGAACATCGTGGAGCTGAAGGAGGTGGTTGTGGGGAACCACCTGGAGAGCATCTTCCTGGTGATGGGTTACTGTGAGCAGGACCTGGCCAGCCTCCTGGAGAATATGCCAACACCCTTCTCGGAGGCTCAGGTCAAGTGCATCGTGCTGCAGGTGCTCCGGGGCCTCCAGTATCTGCACAGGAACTTCATTATCCACAGGGACCTGAAGGTTTCCAACTTGCTCATGACCGACAAGGGTTGTGTGAAGACAGCGGATTTCGGCCTGGCCCGGGCCTATGGTGTCCCAGTAAAGCCAATGACCCCCAAGGTGGTCACTCTCTGGTACCGAGCCCCTGAACTGCTGTTGGGAACCACCACGCAGACCACCAGCATCGACATGTGGGCTGTGGGCTGCATACTGGCCGAGCTGCTGGCGCACAGGCCTCTTCTCCCCGGCACTTCCGAGATCCACCAGATCGACTTGATCGTGCAGCTGCTGGGCACGCCCAGTGAGAACATCTGGCCGGGCTTTTCCAAGCTGCCACTGGTCGGCCAGTACAGCCTCCGGAAGCAGCCCTACAACAACCTGAAGCACAAGTTCCCATGGCTGTCGGAGGCCGGGCTGCGCCTGCTGCACTTCCTGTTCATGTACGACCCTAAGAAAAGGGCGACGGCCGGGGACTGCCTGGAGAGCTCCTATTTCAAGGAGAAGCCCCTACGTCTTCCGATCAGTGGTGTCTGTGAAGGGTGCCGCGAGCCAGGCTGACCAGGCGCCCGGGATCCAGCTCATCCCCTTGGCTGGGAACATCCTCCACTGACTTCCTCCCACTGTCTGCCCTGAACCCACTGCTGCCCCCAGAAAAAGGCCGGGTGACACCGGGGGGCTCCCAGCCCGTGCACCCTGGAAGGGCAGGTCTGGCGGCTCCATCCGTGGCTGCAGGGGTCTCATGTGGTCCTCCTCGCTATGTTGGAAATGTGCAACCACTGCTTCTTGGGAGGAGTGGTGGGTGCAGTCCCCCCGCTGTCTTTGAGTTGTGGTGGACGCTGGCCTGGGATGAGAGGGCCCAGAAGACCTTCGTATCCCCTCTCAGTCGCCCGGGGCTGTCCCGTGCATGGGTTGGCTGTGGGGACCCCAGGTGGGCCTGGCAGGACTCCAGATGAGGACAAGAGGGACAAGGTATGGGGTGGGAGCCACAATTGAGGATACCCCGAGACTACCAGGAGAGCCCTGGGCTGGAGGCTGAGCTGCATCCCTGCTCCCCACATGGAGGACCCAACAGGAGGCCGTGGCTCTGATGCTGAGCGAAGCTATAGGCTCTTGTTGGATAAAAGCTTTTTTAACAGAAAAAAAAAAAAAAAAAAAA


Following is a third CDK10 cDNA sequence (SEQ ID NO: 13).

>gi|32528265|ref|NM_052988.2| Homo sapiens cyclin-dependent kinase (CDC2-like) 10(CDK10), transcript variant 3, mRNAGTCTGCGCCTGCGCGCAAGAGAGGCGGGGCCAGCGCTCGGCATGGCGGAGCCAGATCTGGAGTGCGAGCAGATCCGTCTGAAGTGTATTCGTAAGGAGGGCTTCTTCACGGTGCCTCCGGAACACAGGATCGGGCCCGGGACACCCAGACAGATGAGATTGTCGCACTGAAGAAGGTGCGGATGGACAAGGAGAAGGATGGCATCCCCATCAGCAGCTTGCGGGAGATCACGCTGCTGCTCCGCCTGCGTCATCCGAACATCGTGGAGCTGAAGGAGGTGGTTGTGGGGAACCACCTGGAGAGCATCTTCCTGGTGATGGGTTACTGTGAGCAGGACCTGGCCAGCCTCCTGGAGAATATGCCAACACCCTTCTCGGAGGCTCAGGTCAAGTGCATCGTGCTGCAGGTGCTCCGGGGCCTCCACTATCTGCACAGGAACTTCATTATCCACAGGGACCTGAAGGTTTCCAACTTGCTCATGACCGACAAGGGTTGTGTGAAGACAGGTGGGTGCAACTTGGGCCAGGCCCTGTCCCTAGATGGCACTTGGTGACACACACTCCCCTCTCTGCTGCAGCGGATTTCGGCCTGGCCCGGGCCTATGGTGTCCCAGTAAAGCCAATGACCCCCAAGGTGGTCACTCTCTGGTACCGAGCCCCTGAACTGCTGTTGGGAACCACCACGCAGACCACCAGCATCGACATGTGGGCTGTGGGCTGCATACTGGCCGAGCTGCTGGCGCACAGGCCTCTTCTCCCCGGCACTTCCGAGATCCACCAGATCGACTTGATCGTGCAGCTGCTGGGCACGCCCAGTGAGAACATCTGGCCGGCCTTTTCCAAGCTGCCACTGGTCGGCCAGTACAGCCTCCGGAAGCAGCCCTACAACAACCTGAAGCACAAGTTCCCATGGCTGTCGGAGGCCGGGCTGCGCCTGCTGCACTTCCTGTTCATGTACGACCCTAAGAAAAGGGCGACGGCCGGGGACTGCCTGGAGAGCTCCTATTTCAAGGAGAAGCCCCTACCCTGTGAGCCGGAGCTCATGCCGACCTTTCCCCACCACCGCAACAAGCGGGCCGCCCCAGCCACCTCCGAGGGCCAGAGCAAGCGCTGTAAACCCTGACGGTGGGCCTGGCACACGCCTGTATTCCCACACCAGGTCTTCCGATCAGTGGTGTCTGTGAAGGGTGCCGCGAGCCAGGCTGACCAGGCGCCCGGGATCCAGCTCATCCCCTTCGCTGGGAACATCCTCCACTGACTTCCTCCCACTGTCTGCCCTGAACCCACTGCTGCCCCCAGAAAAAGGCCGGGTGACACCGGGGGGCTCCCAGCCCGTGCACCCTGGAAGGGCAGGTCTGGCGGCTCCATCCGTGGCTGCAGGGGTCTCATGTGGTCCTCCTCGCTATGTTGGAAATGTGCAACCACTGCTTCTTGGGAGGAGTGGTGGGTGCAGTCCCCCCGCTGTCTTTGAGTTGTGGTGGACGCTGGCCTGGGATCAGAGGGCCCAGAAGACCTTCGTATCCCCTCTCAGTCGCCCGGGGCTGTCCCGTGCATGGGTTGGCTGTGGGGACCCCAGGTGGGCCTGGCAGGACTCCAGATGAGGACAACAGGGACAAGGTATGGGGTGGGAGCCACAATTGAGGATACCCCGAGACTACCAGGAGAGCCCTGGGCTGGAGGCTGAGCTGCATCCCTGCTCCCCACATGGAGGACCCAACAGGAGCCCGTGGCTCTGATGCTGAGCGAAGCTATAGGCTCTTGTTGGATAAAAGCTTTTTTAACAGAAAAAAAAAAAAAAAAAAAA


Following is a FPGT cDNA sequence (SEQ ID NO: 14).

>gi|4503776|ref|NM_003838.1| Homo sapiens fucose-1-phosphate guanylyltransferase(FPGT), mRNAGCGTGCTGTGCGGCGCGGTCTCAGGGAAGGTGGGGCTATGGCAGCTGCTAGGGACCCTCCGGAAGTATCGCTGCGAGAAGCCACCCAGCGAAAATTGCGGAGGTTTTCCGAGCTAAGAGGCAAACTTGTAGCACGTGGAGAATTCTGGGACATAGTTGCAATAACAGCGGCTGATGAAAAACAGGAACTTGCTTACAACCAACAGCTGTCAGAAAAGCTGAAAAGAAAGGAGTTACCCCTTGGAGTTCAATATCACGTTTTTGTGGATCCTGCTGGACCCAAAATTGGAAATGGAGGATCAACACTTTGTGCCCTTCAATGTTTGGAAAAGCTATATGGAGATAAATGGAATTCTTTTACCATCTTATTAATTCACTCTGGTGGCTACAGTCAACGACTTCCAAATGCAAGTGCTCTGGGAAAAATTTTCACTGCTTTACCTCTTGGTAACCCCATTTATCAGATGCTAGAATTAAAGCTAGCCATGTACATTGATTTCCCCTTAAATATGAATCCTGGAATTCTGGTTACCTGTGCAGATGATATTGAACTTTATAGTATTGGAGAATTTGAGTTTATTAGGTTTGACAAACCTGGCTTTACTGCTTTAGCTCATCCTTCTAGTTTGACGATAGGTACCACACATGGAGTATTTGTCTTAGATCCTTTTGATGATTTAAAACATAGAGACCTTGAATACAGGTCTTGCCATCGTTTCCTTCATAAGCCCAGCATAGAAAAGATGTATCAGTTTAATGCTGTGTGTAGACCTGGAAATTTTTGTCAACAGGACTTTGCTGGGGGTGACATTGCCGATCTTAAATTAGACTCTGACTATGTCTACACAGATAGCCTATTTTATATGGATCATAAATCAGCAAAAATGTTACTTGCTTTTTATGAAAAAATAGGCACACTGAGCTGTGAAATAGATGCCTATGGTGACTTTCTGCAGGCTTTGGGACCTGGAGCAACTGTGGAGTACACCAGAAACACATCACATGTCATTAAAGAAGAGTCAGAGTTGGTAGAAATGAGGCAGAGAATATTTCATCTTCTTAAAGGAACATCACTAAATGTTGTTGTTCTTAATAACTCCAAATTTTATCACATTGGAACAACCGAAGAATATTTGTTTTACTTTACCTCAGATAACAGTTTAAAGTCAGAGCTCGGCTTACAGTCCATAACTTTTAGTATCTTTCCAGATATACCAGAATGCTCTGGCAAAACATCCTGTATCATTCAAAGCATACTGGATTCAAGATGTTCTGTGGCACCTGGCTCAGTTGTGGAGTATTCCAGATTGGGGCCTGATGTTTCAGTTGGGGAAAACTGCATTATTAGTGGTTCTTACATCCTAACAAAAGCTGCCCTCCCCGCACATTCTTTTGTATGTTCCTTAAGCTTAAAGATGAATAGATGCTTAAAGTATGCAACTATGGCATTTGGAGTGCAAGACAACTTGAAAAAGAGTGTGAAAACATTGTCAGATATAAAGTTACTTCAATTCTTTGGAGTCTGTTTCCTGTCATGCTTAGATGTTTGGAATCTTAAAGTTACAGAGGAACTGTTCTCTGGTAACAAGACATGTCTGAGTTTGTGGACTGCACGCATTTTCCCAGTTTGTTCTTCTTTGAGTGACTCAGTTATAACATCCCTAAAGATGTTAAATGCTGTTAAGAACAAGTCAGCATTCAGCCTGAATAGCTATAAGTTGCTGTCCATTGAAGAAATGCTTATCTACAAAGATGTAGAAGATATGATAACTTACAGGGAACAAATTTTTCTAGAAATCAGTTTAAAAAGCAGTTTGATGTAGAGATATTTTAAATATTGTACACTTTGCCTTTTTGAGTAACATTCCAGAGATAGGTATTTTTGGTAGGCTGTTTCACTGAACTCAGTTAATGAAAACTGTATTAACATAATTGTTGTAGCATAATATTAATAGTGCAAAAGTACATATAAGTCATTTTGATGAAAAATATTCCAAGACTAAGTTGAGAAAAGAGATACTATTTTGGATGTGTATCAGTATTTTTGTTTTTAATAATGATTGATTTGTGGAGCATTGTTTTTTCACATAATTAGTTTTAAAGGTAATTTTCTAAGCATACCTTTGGAATTTTTCCATCTTTTTTGAGGCTTTTGGTCCAGTGAAGTTCTAAGTATTCACTGGCACTTCTCTCCTCAACTGTAATTCTATTTTTAATAATAAAAATGGCATACTGTAGGGTCTTCAGAGTAGTGTAGGAATACTGTAGAAATACTTTTTCAGAAACGAATCCATAGCTGACAAATTCACTCAGTGCCCAATATATTGTGATTATTTTCGTTGATAAAGAACTAGATACAAAGACCTCTGAAATTGATGATAAAATTTGTATCTCATTAATTTTATCAAAATGAAACTAAAAGTACATATGTATTATACACTTGAACATGTGTTTGTATATCTTTAAAATTTTCCTTTATGTCCATTCCATAGGAAGACACACATATGCACACAAAACTCAGTTATCTGTGGGGGAAGTGGTAGTAATAATTGAGATTCATCAATAATCATATAATTTCACTATAGACATTACTTGCATTATCTCCTATCAGTCCTTCTCACAAAACTTCAATTACCAGATGAACTGACTTTAGTTTTCATCTTATTTTTTGTCCACATGCTGGTGATGCTGAGAAACAATAATTGGCCCAAATGCAGACATCAAGAAGCAGGCAGGAAIATGGAAAAACTGAGATAATACACAGGTGATAAACAGTTTCGAGAAAATAAAAATTGGCTAGAATCTCAAGTTATGTGTTTCTATATTGGTATAAGCATATAAGTGAAAAGTCTTATAAGTGTAATAGAGAAAAGATTTGTCAGTGTGTTTTTTTAAATGAAATAACTAGTCTGTGCTACTTTATGTCAATATAAAAATTGGTAAACTAGAAGTAACTTGTCCACAACCCTCAGTTATGATACTTATGTGCGTGTTTTTTTTTCCAAAGTTTTTCCCAAGGAGAAGATACAAATATATGGTAGCTATTGTTTAAAAATGATTGATTTACTTGCAGATTTTTCAGAGGATGTTATGTCTTTGTCATTCATTAAATGTTCAAAATTGTAGTTTTAAAAAAAAAAAAAAA


Following is a CARK cDNA sequence (SEQ ID NO: 15).

>gi|7705747|ref|NM_015978.1| Homo sapiens cardiac ankyrin repeat kinase (CARK), mRNAAATCTTATAACTTGAAGAACTGCCCTGGAGAAAGGAAGAAACTTATAATAAATGGGAAATTATAAATCTAGACCAACCCAAACTTGTACTGATGAATGGAACAAAAAAGTCAGTGAATCATATGTTATCACAATAGAAAGATTAGAAGATGACCTGCAGATCAAGGAAAAAGAACTGACAGAACTAAGGAATATATTTGGCTCTCATCAAGCCTTCAGTAAAGTCAATTTAAATTACCGCACTCAAAATGGGCTGTCTCTACTTCATTTATGTTGCATTTCTGCAGCCAAGAAATCACATATTCGAACTCTTATGTTGAAAGGGCTCCGCCCATCTCGACTGACAAGAAATGGATTTACAGCCTTGCATTTAGCAGTTTACAAGGATAATGCAGAATTGATCACTTCTCTGCTTCACAGTGGAGCTGATATACAGCAGGTTGGATACGGTGGCCTCACTCCCCTCCATATTGCTACAATAGCTGGCCACCTAGAGGCTGCTGATGTGCTGTTGCAACATGGAGCTAATGTCAATATTCAAGATGCAGTTTTTTTCACTCCATTGCATATTGCAGCGTACTATGGACATGAACAGGTAACTCGCCTTCTTTTGAAATTTGGTGCTGATGTAAATGTAAGTGGTGAAGTTGGAGATAGACCCCTCCACCTAGCATCTGCAAAAGGATTCTTGAATATTGCAAAACTCTTGATGGAAGAAGGCAGCAAAGCACATGTGAATGCTCAAGATAATGAAGACCATGTCCCACTCCATTTCTGTTCTCGATTTGGACACCATGATATAGTTAAGTATCTGCTGCAAAGTGATTTGGAAGTTCAACCTCATGTTGTTAATATCTATGGAGATACCCCCTTACACCTGGCATGCTACAATGGCAAATTTGAAGTTGCCAACGAAATCATCCAAATATCAGGAACAGAAAGTCTGACTAAGGAAAACATCTTCAGTGAAACAGCTTTTCATAGTGCTTGTACCTATGGCAAGAGCATTGACCTAGTCAAATTTCTTCTTGATCAGAATGTCATAAACATCAACCACCAAGGAAGGGATGGGCACACTGGATTACACTCTGCTTGCTACCACGGTCACATTCGCCTGGTTCAGTTCTTACTGGATAATGGAGCTGATATGAATCTAGTGGCTTGTGATCCCAGCAGGTCTAGTGGTGAAAAAGATGAGCAGACATGTTTGATGTGGGCTTATGAAAAAGGGCATGATGCCATTGTCACACTCCTGAAGCATTATAAGAGACCACAAGATGAATTGCCCTGTAATGAATATTCTCAGCCTGGAGGAGATGGCTCCTATGTCTCTGTTCCATCACCCTTGGGGAAGATTAAAAGCATGACAAAAGAGAAGGCAGATATTCTCCTCCTAAGAGCTGGATTGCCTTCACATTTCCATCTTCAGCTCTCAGAAATTGAGTTCCATGAGATTATTGGCTCAGGTTCTTTTGGGAAAGTATATAAAGGACGATGCAGAAATAAAATAGTGGCTATAAAACGTTATCGAGCCAATACCTACTGCTCCAAGTCAGATGTGGATATGTTTTGCCGAGAGGTGTCCATTCTCTGCCAGCTCAATCATCCCTGCGTAATTCAGTTTGTGGGTGCTTGCTTGAATGATCCCAGCCAGTTTGCCATTGTCACTCAATACATATCAGGGGGTTCTCTGTTCTCCCTCCTTCATGAGCAGAAGAGGATTCTTGATTTGCAGTCTAAATTAATTATTGCAGTAGATGTTGCCAAAGGCATGGAGTACCTTCACAACCTGACACAGCCARTTATACATCGTGACTTGAACAGTCACAATATTCTTCTCTATGAGGATGGGCATGCTGTGGTGGCAGATTTTGGAGAATCAAGATTTCTACAGTCTCTGGATGAAGACAACATGACAAAACAACCTGGGAACCTCCGTTGGATGGCTCCTGAGGTGTTCACGCAGTGCACTCGGTACACCATCAAAGCAGATGTCTTCAGCTATGCTCTGTGTCTGTGGGAAATTCTCACTGGCGAAATTCCATTCGCTCATCTCAAGCCAGCGGCTGCGGCAGCAGACATGGCTTACCACCACATCAGACCTCCCATTGGCTATTCCATTCCCAAGCCCATATCATCTCTGCTGATACGAGGGTGGAACGCATGTCCTGAAGGAAGACCCGAATTTTCTGAAGTTGTCATGAAGTTAGAAGAGTGTCTCTGCAACATTGAGCTGATGTCTCCTGCATCAAGTAACAGCAGTGGGTCTCTCTCACCTTCTTCTTCTTCTGATTGCCTGGTGAACCGGGGAGGACCTGGCCGGAGTCATGTGGCAGCATTAAGAAGTCGTTTCGAATTGGAATATGCTCTAAATGCAAGGTCCTATGCTGCTTTGTCCCAAAGTGCTGGACAATATTCCTCTCAAGGTCTGTCTTTGGAGGAGATGAAAAGAAGTCTTCAATACACACCCATTGACAAATATGGCTATGTATCCGATCCCATGAGCTCAATGCATTTTCATTCTTGCCGAAATAGTAGCAGCTTTGAGGACAGCAGCTGACAGCATTCGGCGTATACCTARGGAGAGTTTTTTCCCCGAACTGACAGCAACGATTCCAACCACGGCAAGCTGGCTTCCAACTATAACATTTTACTCTCAAAGGTCTCCTTAAATTGGGCTTGTTTTTACTTGTCCTATTTAATTCCCCACTATTAGCAGGCTTTGGATTTGTGCCTAAGGAATAATATGCAAAAGAACCAAGACACAATGTATATGAAGAATTGTTTTTAATTTTGTAAATTAAAAAAAAATTTAGATCGTTACTTGGAAATGGAGCCTAAGTCTGTGGTGGACAGATAATAATTATGTTTTCCTGGGCTGAATTATGTAGACTTGTGTTTGACAGCTATGGGTTTATTTCTTAGAACATTGTTCATTTTCTTTTCTCATTATGTTACTTCTAGTGTTCACCTCTGTGATTAAAGATTCTTTGGTGAAATAG


Following is a PCLO cDNA sequence (SEQ ID NO: 16).

>gi|37538611|ref|XM_168530.2| Homo sapiens piccolo (presynaptic cytomatrix protein)(PCLO), mRNAATGACATCCTCTGGAGCCCAGAAAAAAGTTAAAAGAACTCTGCCAAATCCACCTCCTGAGGAGATTTCCACAGGAACTCAATCCACATTCAGCACAATGGGCACAGTTTCCAGGAGAAGGATCTGCAGAACCAACACAATGGCACGAGCCAAGATTCTCCAGGACATAGACAGAGAGCTTGATCTTGTGGAAAGGGAGTCTGCAAAACTTCGAAAGAAACAAGCAGAGCTTGATGAAGAAGAAAAGGAGATTGATGCTAAGCTACGATACCTGGAAATGGGAATTAACAGGAGGAAAGAGGCCCTATTAAAGGAGAGAGAAAAGAGAGAACGAGCCTACCTCCAGGGAGTAGCTGAGGATCGTGATTACATGTCTGACAGTGAAGTGAGTAGCACAAGACCAACCCGAATAGAAAGTCAGCATGGCATTGAGCGACCAAGAACTGCTCCCCAAACTGAATTCAGCCAGTTTATACCACCACAAACCCAAACAGAATCTCAACTAGTTCCTCCGACAAGTCCTTACACACAATACCAGTACTCTTCCCCTGCTCTTCCTACCCAAGCACCCACCTCATACACTCAACAGTCTCATTTTGAGCAACAAACTTTGTACCATCAGCAAGTTTCACCTTATCAGACTCAGCCAACATTCCAAGCTGTGGCAACAATGTCCTTCACACCTCAAGTTCAACCTACACCAACCCCACAGCCTTCTTATCAGTTACCTTCACAGATGATGGTGATACAACAGAAGCCACGGCAAACTACATTATATTTGGAGCCCAAGATAACCTCAAACTATGAAGTGATTCGCAACCAACCCCTTATGATAGCACCTGTTTCTACGGATAACACATTTGCTGTTTCCCATCTTGGTAGTAAGTACAATAGTTTAGACTTGAGAATAGGTTTGGAGGAAAGAAGTAGCATGGCAAGCAGTCCAATATCAAGCATATCTGCAGATTCTTTCTATGCAGATATTGATCACCATACTCCACGAAATTATGTCCTAATTGACGACATTGGAGAGATCACCAAAGGAACAGCGGCATTAAGCACCGCATTTAGCCTTCATGAAAAGGATCTGTCAAAAACAGACCGTCTCCTTCGAACCACTGAGACACGCCGGTCTCAAGAAGTGACAGATTTCCTAGCACCTTTACAGTCTTCCTCTAGATTGCATAGTTATGTGAAGGCGGAGGAAGACCCAATGGAGGATCCTTACGAGTTAAAGCTTCTGAAACATCAGATTAAACAGGAATTTCGTAGAGGGACAGAGAGCTTAGATCACCTTGCTGGTCTTTCTCATTATTACCATGCTGATACTAGCTACAGACATTTTCCAAAATCTGAGAAGTATAGCATCAGTAGACTCACACTTGAAAAACAAGCAGCAAAACAACTGCCAGCAGCCATACTTTATCAAAAGCAGTCAAAGCATAAGAAATCACTAATTGACCCTAAAATGTCAAAATTTTCACCTATTCAAGAAACTAGAGACCTTGAACCTGATTATTCAAGCTATATGACTTCTAGCACTTCATCTATTGGTGGCATTTCCTCCAGGGCAAGGCTCCTTCAAGATGACATCACTTTTGGCCTCAGAAAAAATATTACAGACCAACAAAAATTTATGGGATCTTCTCTTGGCACAGGACTGGGCACATTAGGAAATACCATACGCTCAGCTCTGCAGGATGAAGCGGATAAGCCATACAGTAGTGGCAGCAGGTCCAGACCTTCCTCCAGACCTTCCTCTGTCTATGGGCTTGATTTATCAATTAAAAGGGATTCTTCTAGCTCTTCCCTAAGACTGAAAGCTCAAGAGGCTGAAGCTCTAGATGTTTCCTTTAGTCATGCATCATCCTCTGCCAGAACTAAGCCGACCAGTTTGCCAATTAGTCAAAGTAGAGGAAGAATACCAATTGTGGCCCAGAATTCTGAAGAAGAAAGCCCACTCAGTCCTGTTGGCCAGCCAATGGGAATGGCCAGGGCTGCAGCTGGACCCCTGCCACCAATATCTGCAGACACCAGGGATCAGTTTGGATCAAGCCACTCATTGCCTGAAGTTCAGCAACACATGAGGGAAGAATCACGGACTCGAGGCTATGACCGTGACATAGCATTCATCATGGATGACTTCCAACATGCCATGTCAGACAGTGAAGCCTATCATCTGCGTCGTGAGGAAACAGATTGGTTTGATAAACCCAGGGAGTCTCGTTTGGAAAATGGACATGGTCTGGACCGAAAACTGCCGGAAAGATTGGTCCACTCTAGACCACTCAGTCAACATCAAGAGCAAATTATACAGATGAACGGGAAAACTATGCACTACATCTTTCCTCACGCAACGATAAAAATAACAAGAGACTCAAAGGATCACACAGTTTCAGGTAATGGATTAGGAATTAGAATTGTGGGTGGTAAAGAAATCCCGGGACATAGTGGAGAAATTCGAGCCTATATTGCCAAGATTCTTCCTGGGGGAAGTGCGGAACAGACCGGGAAGCTTATGGAAGGGATGCAAGTATTGGAATGGAATGGAATTCCCTTGACTTCTAAAACATATGAAGAAGTTCAGAGTATCATTAGTCAGCAAAGTGGGGAACCAGAAATATGTGTAAGACTGGACCTCAATATGCTATCAGATTCTGAAAATTCCCAGCATCTGGAACTTCATGAGCCACCAAAAGCTGTGGATAAGGCGAAATCCCCAGGGGTTGATCCTAAGCAGTTGGCAGCAGAACTCCAGAAGGTTTCACTACAGCAGTCACCGCTGGTTCTGTCATCAGTTGTTGAAAAAGGATCTCATGTTCATTCAGGTCCTACATCAGCAGGATCCAGTTCCGTTCCCAGCCCTGGGCAACCAGGGTCCCCCTCAGTGAGCAAAAAGAAGCACGGCAGCAGCAAGCCTACCGATGGAACAAAGGTTGTCTCTCATCCAATTACAGGAGAAATTCAGCTTCAAATTAACTATGATCTTGGAAATCTCATAATACATATTCTCCAAGCAAGAAATCTTGTTCCTCGAGACAACAATGGTTATTCTGACCCTTTTGTGAAAGTGTACCTTCTTCCAGGGAGAGGTCAAGTCATGGTTGTCCAGAATGCAAGTGCTGAGTACAAGAGAAGGACTAAACATGTCCAGAAA~GTCTTAATCCTGAGTGGAATCAAACAGTAATTTATAAAAGTATTTCCATGGAACAGCTCAAGAAGAAAACACTGGAGGTGACAGTTTGGGATTATGATAGATTTTCATCCAACGACTTCCTTGGGGAGGTATTGATTGATTTATCTAGCACATCTCACCTCGATAACACTCCAAGGTGGTATCCTCTCAAAGAACAGACTGAAAGCATTGATCATGGCAAGTCTCATTCCAGTCAGAGCAGCCAGCAGTCCCCAAAGCCATCTGTTATCAAAAGCAGAAGCCATGGTATCTTCCCTGACCCATCAAAGGACATGCAGGTTCCCACCATTGAGA~ATCCCATAGTAGTCCTGGTAGCTCAAAATCATCATCAGAAGCCCATCTCCGTTCTCATGGACCATCTCCCAGTCAAAGCAAAACCAGCGTCACTCAGACCCACCTGGAAGATGCAGGGGCTGCCATAGCTGCTGCCGAAGCTCCCGTGCAACAACTCCGCATTCAACCAAGTAAAAGACGCAAATAAATTCCTCAGCATGGCAGCTTAATGTTCATCTGTTGCCTTTCTTTCCTGCTGTCCTTTCCTGTTTGCTTTCAGTTTTCAACATCCTCTGCTCACCCTGTTCTCTGTCCCTTTGTCTGTGTAAGAACGATATAAATACATTAATATGCTCTTTCTTATTTAGATTTTTTTTATTTACATAGACTGAAATAAAACTGGCTGTTTCTCCTTTGTTTCCACCATCCATCCAACCTGGCTCATAGCATTTGATACAGTGTCTGTGATGTTTGGAAGCAAAGCAATGTTGTGTGTCCTTTTTGTTTGCGCTTAAATATCTTTTAGAATACAGAAATTATAAGCACAAGTGGCTGCCAGTTTTCCAGGACTTATACCAAAAAAAAAAAAGTGCACACATATAAGTATTTAGAGCAGATTTGCAATTAGATTCTTGTATCTGAAAACACTCAAAAAGTCTAGGCAGAAGATAATGCAACCAACAGTCAACTTCTAGAACCTTGGAGGAAAAGCCATTTCAAGACAGTGCTTTAAAACACTAACAAATTTTGCTTCTGAAATATGGTAGCTTTCCATGTGCAAAAAATTGTAGTCTTCTGAAAAATCTGTGAAGAAAATCTTATTTCTTACATTCTCAAAAAACTACATTAAATTAATATATGTATAAGCATATGTGATAAGGCAATGCAACCTCACTGAGGAGCACAATTGGGTCCATTCTGTTGGATCACACTTTGTATGTGATGACAGATTTTACAGATATTTTTTCTAACCATTGGAAATCAAGATATACTATGTCTGGTAGCTATAAGTCAATCAAAGTATAACAATTGACCTATCTTAACTGCTGTATTTTCATTTCATTTGTGAAAATTTAGTCGATTGATATAATTATATAAAAACTTAATATGTGCAAATATTTCACAAATGTAGGTACCCTGTTACTTCTAAAAGGCATTAGCCAGGCAATGGAGATAGATCATTGTTCTCTCCATCTCCCTAGTCATCATTTTCAAAGAAACAGCCAGTTTTAAACCCCTTTAGTGCTGTGCATGTGGTGTCATTTTGGCATTTGAAAACACAACCTATTTAAATAGAGCAAAAGAGCACTTCTGTTTGCATGAATAACATTTAAGCTGGTTCCATCTGAGGATACATACTCTGGGTTTCCATTTCACCCACACTGCCACAGCAGCACATAAAAGCGGTCAGTCTAATCATGCCAGGAAGCAGCACAGACACAGCATAGCAGGAGTCCTCCCCATTCAAAGAACTCAGTCTGATAATCTGCCTCCACCAGCCAATGGCAACCAAGACCAAAGCCAGCTAGCCCTCAGGAAGGTGATGTCTGATGGACCAGTCAAGCCAGAAGGAGGTGAGCAGAAAGGCCCTGTTCAAAGCACACAGCAAGAACATCCCCAAATTTAAAGCTCTCATCTTTAATTTTTAGAGATTGTCTGATAAAATATGTTTATCCACTTTTTAATGTGCTTTTGTGGGCATGGCATTTCAGGACTTCATCATCACCATGTAAGTCCCAAGATCAGTCCCGTAATATATTGACTACAGTGATTTGTATTTTTCTATGCACTGGGCATCTTTTGACTTTTGTTGGGAAATAGATTAACAATATTCATATTGCAGAATGGAATGCATGCTACTAACCTAATCTGTTCAAGCATGACTATAAATATGTTTCATTGTGATAAAAGTTTAAATTGCTCACAGTTCAAGTTAGCCTACCAGTTGGAATGTGGATCAGTGTTTTTTGCCTATTATTTCAAGTGTAACTTTTAAAAATTTAGTGGATTTTGAAAATTCTTAGAGGAAAGTAAAGGAAAAATTGTTAATGCACTCATTTACCTTTACATGGTGAAAGTTCTCTCTTGATCCTACAAACAGACATTTTCCACTCGTGTTTCCATAGTTGTTAAGTGTATCAGATGTGTTGGGCATGTGAATCTCCAAGTGCCTGTGTAATAAATAAAGTATCTTTATTTCATTCAT2


Following is a REPS2 cDNA sequence (SEQ ID NO: 17).

>gi|4758943|ref|NM_004726.1| Homo sapiens RALBP1 associated Eps domain containing 2(REPS2), MrnaCCGGGAGGAAGCGGCCGCGCGGCAGCTGCGGGGCGTGGGGGTGGTGGTGGCGGCGGCGGTGGTGGCGGCGGCGGCGGCGGCGGCAGCTGAGGCCGAGGAGGCGGTGGCTGTGGCGGACGCAGCAGCACCCCAGCTAGGGATAGGGCTCCGCCGCGCCCCCTTGCTGGCCCCATGGAGGCGGCAGCGGCGGCGGCGGCGGCGGCAGCGGCAGCGGCAGCGGCGGGCGGGGGCTGTGGCTCCGGGCCGCCGCCGCTGCTGCTGAGCGAGGGCGAGCAGCAGTGCTACTCCGAGCTCTTCGCGCGCTGTGCCGGCGCCGCGGGCGGGGGCCCCGGGTCTGGGCCCCCCGAGGCCGCCAGAGTCGCCCCCGGCACGGCCACTGCGGCCGCCGGCCCCGTGGCTGACCTGTTTCGGGCATCGCAGCTGCCCGCCGAGACGCTGCACCAGATCACACAACTGTGTGGTGCAAAGCGGGTTGGTTATTTTGGTCCAACACAGTTTTACATTGCCCTGAAATTAATTGCTGCAGCACAATCTGGCCTCCCGGTACGGATAGAGAGTATTAAATGTGAATTGCCTCTGCCTCGCTTTATGATGTCAAAGAATGATGGTGAGATACGATTTGGGAACCCAGCTGAGCTGCATGGAACTAAGGTTCAGATTCCATATTTAACTACAGAAAAAAATTCCTTCAAAAGAATGGACGATGAGGATAAACAGCAGGAAACACAGTCTCCCACGATGTCACCCCTCGCCTCCCCTCCTTCTTCCCCGCCTCATTACCAGAGGGTGCCCTTGAGCCATGGCTACAGCAAACTGCGGAGCAGCGCAGAACAGATGCATCCAGCACCTTATGAAGCTAGGCAGCCCCTTGTCCAGCCCGAGGGATCCTCATCAGGGGGCCCAGGAACCAAGCCCCTTCGGCATCAGGCTTCCCTTATCCGGTCCTTTTCAGTGGAGAGGGAACTACAGGATAACAGCAGTTACCCCCACGAACCCTGGAGGATAACAGAAGAACAGCGCGAGTACTATGTCAATCAGTTCCGATCCCTTCAGCCACACCCAAGCTCTTTCATTTCAGGTTCTGTGGCCAAGAACTTCTTCACCAAATCAAAGCTTTCCATTCCAGAACTCTCCTATATATGGGAGCTTAGTGATGCTGACTGTGATGGAGCCCTGACCCTGCCTGAGTTCTGTGCTGCGTTTCATCTCATTGTGGCTCGGAAGAACGGCTACCCATTGCCTGAGGGCCTCCCTCCAACTCTGCAGCCAGAATACCTGCAGGCAGCTTTTCCTAAGCCCAAATGGGACTGTCAATTATTTGATTCTTATTCTGAGTCACTGCCGGCAAATCAACAACCTCGTGACTTGAATCGGATGGAGAAGACATCTGTTAAAGACATGGCTGACCTTCCTGTCCCTAACCAGGATGTAACTAGTGATGACAAACAAGCTTTGAAAAGTACTATCAATGAAGCCTTACCAAAGGACGTGTCTGAGGATCCAGCAACTCCCAAGGATTCCAACAGTCTCAAAGCAAGACCAAGATCCAGATCTTACTCTAGCACCTCCATAGAAGAGGCCATGAAAAGGGGCGAGGACCCTCCCACCCCGCCACCTCGGCCACAGAAAACCCATTCCAGAGCCTCCTCCTTGGATCTGAATAAAGTCTTCCAGCCCAGTGTGCCAGCTACCAAGTCAGGATTGTTACCCCCACCACCTGCGCTCCCTCCAAGACCTTGTCCATCACAGTCTGAACAAGTGTCGGAGGCCGAGTTACTCCCACAGCTGACCAGAGCCCCATCCCAGGCTGCAGAAAGTAGTCCAGCAAAGAAGGATGTACTGTATTCTCAGCCACCATCAAAGCCCATTCGTAGGAAATTCAGACCAGAAAACCAAGCTACAGAAAACCAAGAGCCTTCCACTGCTGCAAGTGGGCCAGCTTCTGCGGCAACCATGAAACCGCATCCAACAGTCCAAAAGCAGTCTTCCAAACAGAAGAAGGCCATTCAAACTGCTATCCGCAAAAATAAAGAGGCAAACGCAGTGCTGGCTCGGCTGAACAGTGAGCTCCAGCAGCAGCTCAAGGAGGTTCATCAAGAACGAATTGCATTGGAAAACCAATTGGAACAACTTCGTCCGGTCACTGTGTTGTGACCCCCCCATGGTTCAAGTGACAGTGGGTGACCTTGTCTGCCAAGATCTTTCTTTTGAATGTTTTGAACCCAACTACTTGTCATAGATGTTTGACTGTGTCAAAAGCTGTGAGCAGCAAAATATAATCCATA


Following are NRP1, NID2, ENDO180, CDK10, FPGT, CARK, PCLO and REPS2 amino acid sequences (SEQ ID NO: 18, 19, 20, 21, 22, 23, 24, 25, 26 and 27, respectively). Following is a NRP1 amino acid sequence (SEQ ID NO: 18).

>gi|4505457|ref|NP_003864.1| neuropilin 1 [Homo sapiens]MERGLPLLCAVLALVLAPAGAFRNDECGDTIKIESPGYLTSPGYPHSYHPSEKCEWLIQAPDPYQRIMINFNPHFDLEDRDCKYDYVEVFDGENENGHFRGKFCGKLAPPPVVSSGPFLFIKFVSDYETHGAGFSIRYEIFKRGPECSQNYTTPSGVIKSPGFPEKYPNSLECTYIVFAPKMSEIILEFESFDLEPDSNPPGGMFCRYDRLEIWDGFPDVGPHIGRYCGQKTPGRIRSSSGILSMVFYTDSAIAKEGFSANYSVLQSSVSEDFKCMEALGMESGEIHSDQITASSQYSTNWSAERSRLNYPENGWTPGEDSYREWIQVDLGLLRFVTAVGTQGAISKETKKKYYVKTYKIDVSSNGEDWITIKEGNKPVLFQGNTNPTDVVVAVFPKYLITRFVRIKPATWETGISMRFEVYGCKITDYPCSGMLGMVSGLISDSQITSSNQGDRNWMPENLRLVTSRSGWALPPAPHSYINEWLQIDLGEEKIVRGIIIQGGKHRENKVFMRKFKIGYSNNGSDWKMIMDDSKRKAKSFEGNNNYDTPELRTFPALSTRFIRIYPERATHGGLGLRMELLGCEVEAPTAGPTTPNGNLVDECDDDQANCHSGTGDDFQLTGGTTVLATEKPTVIDSTIQSEFPTYGFNCEFGWGSHKTFCHWEHDNHVQLKWSVLTSKTGPIQDHTGDGNFIYSQADENQKGKVARLVSPVVYSQNSAHCMTFWYHMSGSHVGTLRVKLRYQKPEEYDQLVWMAIGHQGDHWKEGRVLLHKSLKLYQVIFEGEIGKGNLGGIAVDDISINNHISQEDCAKPADLDKKNPEIKIDETGSTPGYEGEGEGDKNISRKPGNVLKTLEPILITIIAMSALGVLLGAVCGVVLYCACWHNGMSERNLSALENYNFELVDGVKLKKDKLNTQSTYSEA


The following Table shows polypeptide domain regions in a NRP1 amino acid sequence.

AAPredictionAccessionstartAADomainmethodidsiteenddescriptionprintsPR00020692708MAM_domainprintsPR00020720731MAM_domainprintsPR00020769783MAM_domainprintsPR00020788801MAM_domainPfamPF0043127138CUB_domainPfamPF00431147262CUB_domainPfamPF00754278421FA58_CPfamPF00754434580FA58_CPfamPF00629650811MAM_domainprositePS00740690730MAM_domainprositePS01285314343FA58_CprositePS01285470502FA58_CprositePS01286408424FA58_CprositePS01286566583FA58_CpfscanPS0118027141CUB_domainpfscanPS01180147256CUB_domain


Following is a NID2 amino acid sequence (SEQ ID NO: 19).

>gi|33469989|ref|NP_031387.2| nidogen 2; osteonidogen [Homo sapiens]MEGDRVAGRPVLSSLPVLLLLQLLMLRAAALHPDELFPHGESWGDQLLQEGDDESSAVVKLANPLHFYEARFSNLYVGTNGIISTQDFPRETQYVDYDFPTDFPAIAPFLADIDTSHGRGRVLYREDTSPAVLGLAARYVRAGFPRSARFTPTHAFLATWEQVGAYEEVKRGALPSGELNTFQAVLASDGSDSYALFLYPANGLQFLGTRPKESYNVQLQLPARVGFCRGEADDLKSEGPYFSLTSTEQSVKNLYQLSNLGIPGVWAFHIGSTSPLDNVRPAAVGDLSAAHSSVPLGRSFSHATALESDYNEDNLDYYDVNEEEAEYLPGEPEEALNGHSSIDVSFQSKVDTKPLEESSTLDPHTKEGTSLGEVGGPDLKGQVEPWDERETRSPAPPEVDRDSLAPSWETPPPYPENGSIQPYPDGGPVPSEMDVPPAHPEEEIVLRSYPASGHTTPLSRGTYEVGLEDNIGSNTEVFTYNAANKETCEHNHRQCSRHAFCTDYATGFCCHCQSKFYGNGKHCLPEGAPHRVNGKVSGHLHVGHTPVHFTDVDLHAYIVGNDGRAYTAISHIPQPAAQALLPLTPIGGLFGWLFALEKPGSENGFSLAGAAFTHDMEVTFYPGEETVRITQTAEGLDPENYLSIKTNIQGQVPYVPANFTAHISPYKELYHYSDSTVTSTSSRDYSLTFGANQTWSYRIHQNITYQVCRHAPRHPSFPTTQQLNVDRVFALYNDEERVLRFAVTNQIGPVKEDSDPTPGNPCYDGSHMCDTTARCHPGTGVDYTCECASGYQGDGRNCVDENECATGFHRCGPNSVCINLPGSYRCECRSGYEFADDRHTCILITPPANPCEDGSHTCAPAGQARCVHHGGSTFSCACLPGYAGDGHQCTDVDECSENRCHPAATCYNTPGSFSCRCQPGYYGDGFQCIPDSTSSLTPCEQQQRHAQAQYAYPGARFHIPQCDEQGNFLPLQCHGSTGFCWCVDPDGHEVPGTQTPPGSTPPHCGPSPEPTQRPPTICERWRENLLEHYGGTPRDDQYVPQCDDLGHFLPLQCHGKSDFCWCVDKDGREVQGTRSQPGTTPACIPTVAPPMVRPTPRPDVTPPSVGTFLLYTQGQQIGYLPLNGTRLQKDAAKTLLSLHGSIIVGIDYDCRERMVYWTDVAGRTISRAGLELGAEPETIVNSGLISPEGLAIDHIRRTMYWTDSVLDKIESALLDGSERKVLFYTDLVNPRAIAVDPIRGNLYWTDWNREAPKIETSSLDGENRRILINTDIGLPNGLTFDPFSKLLCWADAGTKKLECTLPDGTGRRVIQNNLKYPFSIVSYADHFYHTDWRRDGVVSVNKHSGQFTDEYLPEQRSHLYGITAVYPYCPTGRK


The following Table shows domain regions in a NID2 amino acid sequence.

AAPredictionAccessionstartAADOMAINmethodidsiteendDESCRIPTIONPfamPF00008763799EGF_likePfamPF00008805842EGF_likePfamPF00008852890EGF_likePfamPF00008896929EGF_likePfamPF000869401005Thyroglobulin_1PfamPF0008610191084Thyroglobulin_1PfamPF0005811541196Ldl_receptor_repPfamPF0005811981239Ldl_receptor_repPfamPF0005812411284Ldl_receptor_repPfamPF0005812861326Ldl_receptor_repprositePS00010501512Asx_hydroxylprositePS00010818829Asx_hydroxylprositePS00010907918Asx_hydroxylprositePS00484959988Thyroglobulin_1prositePS0048410391068Thyroglobulin_1prositePS01186786799EGF_likeprositePS01186827842EGF_likeprositePS01186877890EGF_likeprositePS01186916929EGF_likeprositePS01187801827EGF_CaprositePS01187892916EGF_Ca


Following is a ENDO180 amino acid sequence (SEQ ID NO: 20).

>gi|5174485|ref|NP_006030.1| mannose receptor, C type 2MGPGRPAPAPWPRHLLRCVLLLGCLHLGRPGAPGDAALPEPNVFLIFSHGLQGCLEAQGGQVRVTPACNTSLPAQRWKWVSRNRLFNLGTMQCLGTGWPGTNTLTASLGMYECDREALNLRWHCRTLGDQLSLLLGARTSNISKPGTLERGDQTRSGQWRIYGSEEDLCALPYHEVYTIQGNSHGKPCTIPFKYDNQWFHGCTSTGREDGHLWCATTQDYGKDERWGFCPIKSNDCETFWDKDQLTDSCYQFNFQSTLSWREAWASCEQQGADLLSITEIHEQTYINGLLTGYSSTLWIGLNDLDTSGGWQWSDNSPLKYLNWESDQPDNPSEENCGVIRTESSGGWQNRDCSIALPYVCKKKPNATAEPTPPDRWANVKVECEPSWQPFQGHCYRLQAEKRSWQESKKACLRGGGDLVSIHSMAELEFITKQIKQEVEELWIGLNDLKLQMNFEWSDGSLVSFTHWHPFEPNNFRDSLEDCVTIWGPEGRWNDSPCNQSLPSICKKAGQLSQGAAEEDHGCRKGWTWHSPSCYWLGEDQVTYSEARRLCTDHGSQLVTITNRFEQAFVSSLIYNWEGEYFWTALQDLNSTGSFFWLSGDEVMYTHWNRDQPGYSRGGCVALATGSAMGLWEVKNCTSFRARYICRQSLGTPVTPELPGPDPTPSLTGSCPQGWASDTKLRYCYKVFSSERLQDKKSWVQAQGACQELGAQLLSLASYEEEHFVANMLNKIFGESEPEIHEQHWFWIGLNRRDPRGGQSWRWSDGVGFSYHNFDRSRHDDDDIRGCAVLDLASLQWVAMQCDTQLDWICKIPRGTDVREPDDSPQGRREWLRFQEAEYKFFEHHSTWAQAQRICTWFQAELTSVHSQAELDFLSHNLQKFSRAQEQHWWIGLHTSESDGRFRWTDGSIINFISWAPGKPRPVGKDKKCVYMTASREDWGDQRCLTALPYICKRSNVTKETQPPDLPTTALGGCPSDWIQFLNKCFQVQGQEPQSRVKWSEAQFSCEQQEAQLVTITNPLEQAFITASLPNVTFDLWIGLHASQRDFQWVEQEPLMYANWAPGEPSGPSPAPSGNKPTSCAVVLHSPSAHFTGRWDDRSCTEETHGFICQKGTDPSLSPSPAALPPAPGTELSYLNGTFRLLQKPLRWHDALLLCESHNASLAYVPDPYTQAFLTQAARGLRTPLWIGLAGEEGSRRYSWVSEEPLNYVGWQDGEPQQPGGCTYVDVDGAWRTFfSCDTKLQGAVCGVSSGPPPPRRISYHGSCPQGLADSAWJPFREHCYSFHMELLLGHKEARQRCQRAGGAVLSILDEMENVFVWEHLQSYEGQSRGAWLGMNFNPKGGTLVWQDNTAVNYSNWGPPGLGPSMLSHNSCYWIQSNSGLWRPGACTNITMGVVCKLPRAEQSSFSPSALPENPAALVVVLMAVLLLLALLTAALILYRRRQSIERGAFEGARYSRSSSSPTEATEKNILVSDMEMNEQQE


The following Table shows domain regions in an ENDO180 amino acid sequence.

AAPredictionAccessionstartAADOMAINmethodidsiteendDESCRIPTIONprintsPR00356381393AntifreezeIIprintsPR00356393410AntifreezeIIprintsPR00356411428AntifreezeIIprintsPR00356441457AntifreezeIIprintsPR00356491504AntifreezeIIprintsPR00013184193FN_Type_IIprintsPR00013195207FN_Type_IIprintsPR00013212227FN_Type_IIPfamPF00040187228FN_Type_IIPfamPF00059255361Lectin_CPfamPF00059399506Lectin_CPfamPF00059538646Lectin_CPfamPF00059693810Lectin_CPfamPF00059842952Lectin_CPfamPF000599931109Lectin_CPfamPF0005911421245Lectin_CPfamPF0005912841395Lectin_CprositePS00023187228FN_Type_IIprositePS0021312191232Lipocln_cytFABPprositePS00615335359Lectin_CprositePS00615481504Lectin_CprositePS00615927950Lectin_CpfscanPS5023154121Ricin_B_lectin


Following is a first CDK10 amino acid sequence (SEQ ID NO: 21).

>gi|32528263|ref|NP_003665.2| cyclin-dependent kinase 10 isoform 1; serine/threonineprotein kinase PISSLRE; CDC2-related protein kinase; cell division protein kinase 10;cyclin-dependent kinase related protein [Homo sapiens]MGRCRSVKEFEKLNRIGEGTYGIVYRARDTQTDEJVALKKVRMDKEKDGIPISSLREITLLLRLRHPNIVELKEVVVGNHLESIFLVMGYCEQDLASLLENMPTPFSEAQVKCIVLQVLRGLQYLHRNFIIHRDLKVSNLLMTDKGCVKTADFGLARAYGVPVKPMTPKVVTLWYRAPELLLGTFLTTQTTSIDMWAVGCILAELLAHRPLLPGTSEIHQIDLIVQLLGTPSENIWPGFSKLPLVGQYSLRKQPYNNLKHKFPWLSEAGLRLLHFLFMYDPKKRATAGDCLESSYFKEKPLPCEPELMPTFPHHRNKRAAPATSEGQSKRCKIP


Following is a third CDK10 amino acid sequence (SEQ ID NO: 23).

>gi|32528266|ref|NP_443714.2| cyclin-dependent kinase 10 isoforra 3; serine/threonineprotein kinase PISSLRE; CDC2-related protein kinase; cell division protein kinase 10;cyclin-dependent kinase related protein [Homo sapiens]MDKEKDGIPISSLREITLLLRLRHPNIVELKEVVVGNHLESIFLVMGYCEQDLASLLENMPTPFSEAQVKCIVLQVLRGLQYLHRNFIIHRDLKVSNLLMTDKGCVKTGGCNLGQALSLDGTW


Following is a FPGT amino acid sequence (SEQ ID NO: 24).

>gi|4503777|ref|NP_003829.1| fucose-1-phosphate guanyltransferase; GDP-beta-L-fucosepyrophosphorylase [Homo sapiens]MAAARDPPEVSLREATQRKLRRFSELRGKLVARGEFWDIVAITAADEKQELAYNQQLSEKLKRKELPLGVQYHVFVDPAGAKIGNGGSTLCALQCLEKLYGDKWNSFTILLIHSGGYSQRLPNASALGKIFTALPLGNPIYQMLELKLAMYIDFPLNMNPGILVTCADDIELYSIGEFEFIRFDKPGFTALAHPSSLTIGTTHGVFVLDPFDDLKHRDLEYRSCHRFLHKPSIEKMYQFNAVCRPGNFCQQDFAGGDIADLKLDSDYVYTDSLFYMDHKSAKMLLAFYEKIGTLSCEIDAYGDFLQALGPGATVEYTRNTSHVIKEESELVEMRQRIFHLLKGTSLNVVVLNNSKFYHIGTTEEYLFYFTSDNSLKSELGLQSITFSIFPDLPECSGKTSCIIQSILDSRCSVAPGSVVEYSRLGPDVSVGENCIISGSYILTKAALPAHSFVCSLSLKMNRCLKYATMAFGVQDNLKKSXTKTLSDIKLLQFFGVCFLSCLDVWNLKVTEELFSGNKTCLSLWTARIFPVCSSLSDSVITSLKMLNAVKNKSAFSLNSYKLLSIEEMLIYKDVEDMITYREQIFLEISLKSSLM


Following is a CARK amino acid sequence (SEQ ID NO: 25).

>gi|7705748|ref|NP_057062.1| cardiac ankyrin repeat kinase [Homo sapiens]MGNYKSRPTQTCTDEWKKKVSESYVITIERLEDDLQIKEKELTELRMFGSDEAFSKVNLNYRTENGLSLLHLCCICGGKKSHIRTLMLKGLRPSRLTRNGFTALHLAVYKDNAELITSLLHSGADIQQVGYGGLTALHIATLAGHLEAADVLLQHGANVNIQDAVFFTPLHJAAYYGHEQVTRLLLKFGADVNVSGEVGDRPLHLASAKGFLNIAKLLMEEGSKADVNAQDNEDHVPLHFCSRFGHHDIVKYLLQSDLEVQPHVVNIYGDTPLHLACYNGKFEVAKEIIQISGTESLTKENIFSETAFHSACTYGKSIDLVKFLLDQNVININHQGRDGHTGLHSACYHGHIRLVQFLLDNGADMNLVACDPSRSSGEKDEQTCLMWAYEKGHDAIVTLLKHYKRPQDELPCNEYSQPGGDGSYVSVPSPLGKIKSMTKEKADILLLRAGLPSHFHLQLSEIEFHEIIGSGSFGKVYKGRCRNKIVAIKRYRANTYCSKSDVDMFCREVSILCQLNHPCVIQFVGACLNDPSQFAIVTQYISGGSLFSLLHEQKRILDLQSKLIJAVDVAKGMEYLHNLTQPIIHRDLNSHNILLYEDGHAVVADFGESRFLQSLDEDNMTKQPGNLRWMAPEVFTQCTRYTIKADVFSYALCLWEILTGEIPFAHLKPAAAAADMAYHHIRPPIGYSIPKPISSLLIRGWNACPEGRPEFSEVVMKLEECLCNIELMSPASSNSSGSLSPSSSSDCLVNRGGPGRSHVAALRSRFELEYALNARSYAALSQSAGQYSSQGLSLEEMKRSLQYTPIDKYGYVSDPMSSMHFHSCRNSSSFEDSS


Following is a PCLO amino acid sequence (SEQ ID NO: 26).

MTSSGAQKKVKRTLPNPPPEEISTGTQSTFSTMGTVSRRRICRTNTMARAKILQDIDRELDLVERESAKLRKKQAELDEEEKEIDAKLRYLEMGINRRKEALLKEREKRERAYLQGVAEDRDYMSDSEVSSTRPTRIESQHGIERPRTAPQTEFSQFIPPQTQTESQLVPPTSPYTQYQYSSPALPTQAPTSYTQQSHFEQQTLYHQQVSPYQTQPTFQAVATMSFTPQVQPTPTPQPSYQLPSQMMVIQQKPRQTTLYLEPKITSNYEVIRNQPLMIAPVSTDNTFAVSHLGSKYNSLDLRIGLEERSSMASSPISSISADSFYADIDHHTPRNYVLIDDIGEITKGTAALSTAFSLHEKDLSKTDRLLRTTETRRSQEVTDFLAPLQSSSRLHSYVKAEEDPMEDPYELKLLKHQIKQEFRRGTESLDHLAGLSHYYHADTSYRHFPKSEKYSISRLTLEKQAAKQLPAAILYQKQSKHKKSLIDPKMSKFSPIQESRDLEPDYSSYMTSSTSSIGGISSRARLLQDDITFGLRKNITDQQKFMGSSLGTGLGTLGNTIRSALQDEADKPYSSGSRSRPSSRPSSVYGLDLSIKRDSSSSSLRLKAQEAEALDVSFSHASSSARTKPTSLPISQSRGRIPIVAQNSEEESPLSPVGQPMGMARAAAGPLPPISADTRDQFGSSHSLPEVQQHMREESRTRGYDRDIAFIMDDFQHAMSDSEAYHLRREETDWFDKPRESRLENGHGLDRKLPERLVHSRPLSQHQEQIIQMNGKTMHYIFPHARIKITRDSKDHTVSGNGLGIRLVGGKEIPGHSGEIGAYIAKILPGGSAEQTGKIMEGMQVLEWNGIPLTSKTYEEVQSIISQQSGEAEICVRLDLNMLSDSENSQHLELHEPPKAVDKAKSPGVDPKQLAAELQKVSLQQSPLVLSSVVEKGSHVHSGPTSAGSSSVPSPGQPGSPSVSKKKHGSSKPTDGTKVVSHPITGEIQLQINYDLGNLIIHILQARNLVPRDNNGYSDPFVKVYLLPGRGQVMVVQNASAEYKRRTKHVQKSLNPEWNQTVJYKSISMEQLKKKTLEVTVWDYDRFSSNDFLGEVLIDLSSTSHLDNTPRWYPLKEQTESIDHGKSHSSQSSQQSPKPSVIKSRSHGWPDPSKDMQVPTIEKSHSSPGSSKSSSEGHLRSHGPSRSQSKTSVTQTHLEDAGAAIAAAEAAVQQLRIQPSKRRK


Following is a REPS2 amino acid sequence (SEQ ID NO: 27)

>gi|4758944|ref|NP_004717.1| RALBP1 associated Eps domain containing 2; partner ofRal-binding protein 1 [Homo sapiens]MMSKNDGELRFGNPAELHGTKVQIPYLTTEKNSFKRMDDEDKQQETQSPTMSPLASPPSSPPHYQRVPLSHGYSKLRSSAEQMHPAPYEARQPLVQPEGSSSGGPGTKPLRHQASLIRSFSVERELQDNSSYPDEPWRITEEQREYYVNQFRSLQPDPSSFISGSVAKNFFTKSKLSIPELSYIWELSDADCDGALTLPEFCAAFHLIVARKNGYPLPEGLPPTLQPEYLQAAFPKPKWDCQLFDSYSESLPANQQPRDLNRMEKTSVKDMADLPVPNQDVTSDDKQALKSTINEALPKDVSEDPATPKDSNSLKARPRSRSYSSTSIEEAMKRGEDPPTPPPRPQKTHSRASSLDLNKVFQPSVPATKSGLLPPPPALPPRPCPSQSEQVSEAELLPQLSRAPSQAAESSPAKKDVLYSQPPSKPIRRKFRPENQATENQEPSTAASGPASAATMKPHPTVQKQSSKQKKAIQTAIRKNKEANAVLARLNSELQQQLKEVHQERIALENQLEQLRPVTVL


Modifications may be made to the foregoing without departing from the basic aspects of the invention. Although the invention has been described in substantial detail with reference to one or more specific embodiments, those of skill in the art will recognize that changes may be made to the embodiments specifically disclosed in this application, yet these modifications and improvements are within the scope and spirit of the invention, as set forth in the claims which follow. Also, citation of the above publications or documents is not intended as an admission that any of the foregoing is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. U.S. patents and other publications and documents referenced are incorporated herein by reference.

Claims
  • 1. A method for identifying a subject at risk of melanoma, which comprises detecting the presence or absence of a polymorphic variation associated with melanoma at one or more positions selected from the group consisting of rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057, rs5784335, rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907, rs8072519, rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6, rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350, position 206 in SEQ ID NO: 5, rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606 and position 25216 in SEQ ID NO: 7; whereby the presence of the one or more polymorphic variations is indicative of the subject being at risk of melanoma.
  • 2. The method of claim 1, which further comprises obtaining the nucleic acid sample from the subject.
  • 3. The method of claim 1, wherein a polymorphic variation is detected at one or more positions selected from the group consisting of rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057 and rs5784335.
  • 4. The method of claim 1, wherein a polymorphic variation is detected at one or more positions selected from the group consisting of rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907 and rs8072519.
  • 5. The method of claim 1, wherein a polymorphic variation is detected at one or more positions selected from the group consisting of rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6 and position 65039 in SEQ ID NO: 6.
  • 6. The method of claim 1, wherein a polymorphic variation is detected at one or more positions selected from the group consisting of rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350 and position 206 in SEQ ID NO: 5.
  • 7. The method of claim 1, wherein a polymorphic variation is detected at one or more positions selected from the group consisting of rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606 and position 25216 in SEQ ID NO: 7.
  • 8. The method of claim 1, wherein one or more polymorphic variations are detected at one or more positions in linkage disequilibrium with a polymorphic variation at one or more of the positions in claim 1.
  • 9. The method of claim 1, wherein detecting the presence or absence of the one or more polymorphic variations comprises: hybridizing an oligonucleotide to the nucleic acid sample, wherein the oligonucleotide is complementary to a nucleotide sequence in the nucleic acid and hybridizes to a region adjacent to the polymorphic variation; extending the oligonucleotide in the presence of one or more nucleotides, yielding extension products; and detecting the presence or absence of a polymorphic variation in the extension products.
  • 10. The method of claim 1, wherein the subject is a human.
  • 11. A method for identifying a polymorphic variation associated with melanoma proximal to an incident polymorphic variation associated with melanoma, which comprises: identifying a polymorphic variation proximal to the incident polymorphic variation associated with melanoma, wherein the incident polymorphic variation is at a position selected from the group consisting of rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057, rs5784335, rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907, rs8072519, rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6, rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350, position 206 in SEQ ID NO: 5, rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606 and position 25216 in SEQ ID NO: 7; and determining the presence or absence of an association of the proximal polymorphic variant with melanoma.
  • 12. The method of claim 11, wherein the proximal polymorphic variation is within a region between about 5 kb 5′ of the incident polymorphic variation and about 5 kb 3′ of the incident polymorphic variation.
  • 13. The method of claim 11, which further comprises determining whether the proximal polymorphic variation is at a position in linkage disequilibrium with the incident polymorphic variation.
  • 14. The method of claim 11, which further comprises identifying a second polymorphic variation proximal to the identified proximal polymorphic variation associated with melanoma and determining if the second proximal polymorphic variation is associated with melanoma.
  • 15. The method of claim 14, wherein the second proximal polymorphic variant is within a region between about 5 kb 5′ of the incident polymorphic variation and about 5 kb 3′ of the proximal polymorphic variation associated with melanoma.
  • 16. An isolated nucleic acid which comprises a portion of or all of a nucleotide sequence in SEQ ID NO: 3, 4, 5, 6 and/or 7 and comprises one or more polymorphic variations selected from the group consisting of an adenine at position 16727 in SEQ ID NO: 3, a guanine at position 36589 in SEQ ID NO: 4, a thymine at position 44544 in SEQ ID NO: 4, a guanine at position 48304 in SEQ ID NO: 4, a guanine at position 53843 in SEQ ID NO: 6, a guanine at position 63624 in SEQ ID NO: 6, an adenine-thymine deletion at position 65039 in SEQ ID NO: 6, a guanine at position 206 in SEQ ID NO: 5, an adenine at position 9261 in SEQ ID NO: 5, a thymine at position 37592 in SEQ ID NO: 5, a guanine at position 10537 in SEQ ID NO: 7, a cytosine at position 25216 in SEQ ID NO: 7 and a thymine at position 119368 in SEQ ID NO: 7.
  • 17. The isolated nucleic acid of claim 16, which comprises an adenine at position 16727 in SEQ ID NO: 3.
  • 18. The isolated nucleic acid of claim 16, which comprises a polymorphic variation selected from the group consisting of a guanine at position 36589 in SEQ ID NO: 4, a thymine at position 44544 in SEQ ID NO: 4, a guanine at position 48304 in SEQ ID NO: 4.
  • 19. The isolated nucleic acid of claim 16, which comprises a polymorphic variation selected from the group consisting of a guanine at position 53843 in SEQ ID NO: 6, a guanine at position 63624 in SEQ ID NO: 6, an adenine-thymine deletion at position 65039 in SEQ ID NO: 6.
  • 20. The isolated nucleic acid of claim 16, which comprises a polymorphic variation selected from the group consisting of a guanine at position 206 in SEQ ID NO: 5, an adenine at position 9261 in SEQ ID NO: 5, a thymine at position 37592 in SEQ ID NO: 5.
  • 21. The isolated nucleic acid of claim 16, which comprises a polymorphic variation selected from the group consisting of a guanine at position 10537 in SEQ ID NO: 7, a cytosine at position 25216 in SEQ ID NO: 7 and a thymine at position 119368 in SEQ ID NO: 7.
  • 22. An oligonucleotide comprising a nucleotide sequence complementary to a portion of the nucleotide sequence of claim 16, wherein the 3′ end of the oligonucleotide is adjacent to a polymorphic variation.
  • 23. A microarray comprising an isolated nucleic acid of claim 16 linked to a solid support.
  • 24. An isolated polypeptide encoded by the isolated nucleic acid sequence of claim 16.
  • 25. A method of genotyping a nucleic acid at a position selected from the group consisting of position 16727 in SEQ ID NO: 3, position 36589 in SEQ ID NO: 4, position 44544 in SEQ ID NO: 4, position 48304 in SEQ ID NO: 4, position 53843 in SEQ ID NO: 6, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6, position 206 in SEQ ID NO: 5, position 9261 in SEQ ID NO: 5, position 37592 in SEQ ID NO: 5, position 10537 in SEQ ID NO: 7, position 25216 in SEQ ID NO: 7 and position 119368 in SEQ ID NO: 7.
  • 26. The method of claim 25, which comprises genotyping the nucleic acid at position 16727 in SEQ ID NO: 3.
  • 27. The method of claim 25, which comprises genotyping the nucleic acid at a position selected from the group consisting of position 36589 in SEQ ID NO: 4, position 44544 in SEQ ID NO: 4 and position 48304 in SEQ ID NO: 4.
  • 28. The method of claim 25, which comprises genotyping the nucleic acid at a position selected from the group consisting of position 53843 in SEQ ID NO: 6, position 63624 in SEQ ID NO: 6 and position 65039 in SEQ ID NO: 6.
  • 29. The method of claim 25, which comprises genotyping the nucleic acid at a position selected from the group consisting of position 206 in SEQ ID NO: 5, position 9261 in SEQ ID NO: 5 and position 37592 in SEQ ID NO: 5.
  • 30. The method of claim 25, which comprises genotyping the nucleic acid at a position selected from the group consisting of position 10537 in SEQ ID NO: 7, position 25216 in SEQ ID NO: 7 and position 119368 in SEQ ID NO: 7.
  • 31. A method for identifying a candidate molecule that modulates cell proliferation, which comprises: (a) introducing a test molecule to a system which comprises one or more cells and a a nucleic acid comprising one or more polymorphic variations at one or more positions selected from the group consisting of rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057, rs5784335, rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907, rs8072519, rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6, rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350, position 206 in SEQ ID NO: 5, rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606 and position 25216 in SEQ ID NO: 7; and (b) determining whether cell proliferation is increased or decreased compared to a system not containing the test molecule, whereby a increased or decreased cell proliferation identifies the test molecule as a candidate molecule that modulates cell proliferation.
  • 32. The method of claim 31, wherein the system is an animal.
  • 33. The method of claim 31, wherein the system is one or more cells.
  • 34. A method for treating melanoma in a subject, which comprises administering a candidate molecule identified by the method of claim 31 to a subject in need thereof, whereby the candidate molecule treats melanoma in the subject.
  • 35. A method for treating melanoma in a subject or preventing melanoma in a subject, which comprises detecting the presence or absence of one or more polymorphic variations at one or more positions selected from the group consisting of rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057, rs5784335, rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907, rs8072519, rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6, position 65039 in SEQ ID NO: 6, rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350, position 206 in SEQ ID NO: 5, rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606 and position 25216 in SEQ ID NO: 7; and administering a melanoma treatment or administering a melanoma preventative to a subject in need thereof based upon the presence or absence of the one or more polymorphic variations in the nucleic acid sample.
  • 36. The method of claim 35, wherein the one or more polymorphic variations is at one or more positions selected from the group consisting of rs6481845, rs10643659, rs7475391, rs7475394, rs7475396, rs9418057 and rs5784335.
  • 37. The method of claim 35, wherein the one or more polymorphic variations is at one or more positions selected from the group consisting of rs2429390, rs7209331, rs4968706, rs8072984, rs9896444, rs4968617, rs7350907 and rs8072519.
  • 38. The method of claim 35, wherein a polymorphic variation is at position rs8404.
  • 39. The method of claim 35, wherein the one or more polymorphic variations is at one or more positions selected from the group consisting of rs6467914, rs3064869, rs10555175, rs6956848, position 63624 in SEQ ID NO: 6 and position 65039 in SEQ ID NO: 6.
  • 40. The method of claim 35, wherein the one or more polymorphic variations is at one or more positions selected from the group consisting of rs489941, rs496008, rs4650251, rs531412, rs3765654, rs6660350 and position 206 in SEQ ID NO: 5.
  • 41. The method of claim 35, wherein the one or more polymorphic variations is at one or more positions selected from the group consisting of rs5924619, rs5924564, rs4828523, rs5924569, rs2052763, rs1549529, rs5924573, rs4240072, rs5924580, rs4828544, rs5969758, rs1965051, rs1426800, rs1365529, rs5924545, rs5969761, rs6629202, rs5924584, rs1365530, rs6527769, rs2382813, rs2033120, rs2162394, rs1346246, rs1365527, rs4828524, rs5924588, rs5969764, rs5924589, rs5924590, rs5924593, rs4828547, rs5969768, rs4828526, rs5924595, rs5924549, rs5969769, rs4007744, rs2891073, rs5924599, rs1549531, rs5924601, rs5901609, rs5969739, rs5924602, rs1426798, rs4828550, rs5924603, rs1807947, rs2382814, rs5924606 and position 25216 in SEQ ID NO: 7.
  • 42. The method of claim 35, which further comprises extracting and analyzing a tissue biopsy sample from the subject.
  • 43. The method of claim 35, wherein the treatment is one or more selected from the group consisting of administering cisplatin, administering carmustine, administering vinblastine, administering vincristine, administering bleomycin, administering a combination of the foregoing, and surgery.
  • 44. The method of claim 35, wherein the preventative reduces ultraviolet (UV) light exposure to the subject.
RELATED PATENT APPLICATIONS

This patent application claims the benefit of provisional patent application 60/489,703 filed Jul. 23, 2003, having attorney docket number 524593004001. This patent application also claims the benefit of U.S. patent application Ser. Nos. 10/703,817, 10/704,513 and 10/703,789, each filed on Nov. 6, 2003, and having attorney docket no. 524592006100, 524592006200 and 524592006300, respectively. Each of the above-identified patent applications names Richard B. Roth et al. as inventors and is hereby incorporated herein by reference in its entirety, including all drawings and cited documents.

Provisional Applications (1)
Number Date Country
60489703 Jul 2003 US