Interleukin-3 (IL-3) multiple mutation polypeptides

FIELD OF THE INVENTION

The present invention relates to mutants or variants of human interleukin-3 (hIL-3) which contain multiple amino acid substitutions and which may have portions of the native hIL-3 molecule deleted. These hIL-3 multiple mutation polypeptides retain one or more activities of native hIL-3 and may also show improved hematopoietic cell-stimulating activity and/or an improved activity profile which may include reduction of undesirable biological activities associated with native hIL-3.

BACKGROUND OF THE INVENTION

Colony stimulating factors (CSFs) which stimulate the differentiation and/or proliferation of bone marrow cells have generated much interest because of their therapeutic potential for restoring depressed levels of hematopoietic stem cell-derived cells. CSFs in both human and murine systems have been identified and distinguished according to their activities. For example, granulocyte-CSF (G-CSF) and macrophage-CSF (M-CSF) stimulate the in vitro formation of neutrophilic granulocyte and macrophage colonies, respectively while GM-CSF and interleukin-3 (IL-3) have broader activities and stimulate the formation of both macrophage, neutrophilic and eosinophilic granulocyte colonies. IL-3 also stimulates the formation of mast, megakaryocyte and pure and mixed erythroid colonies.

Because of its ability to stimulate the proliferation of a number of different cell types and to support the growth and proliferation of progenitor cells, IL-3 has potential for therapeutic use in restoring hematopoietic cells to normal amounts in those cases where the number of cells has been reduced due to diseases or to therapeutic treatments such as radiation and chemotherapy.

Interleukin-3 (IL-3) is a hematopoietic growth factor which has the property of being able to promote the survival, growth and differentiation of hematopoietic cells. Among the biological properties of IL-3 are the ability (a) to support the growth and differentiation of progenitor cells committed to all, or virtually all, blood cell lineages; (b) to interact with early multipotential stem cells; (c) to sustain the growth of pluripotent precursor cells; (d) to stimulate proliferation of chronic myelogenous leukemia (CML) cells; (e) to stimulate proliferation of mast cells, eosinophils and basophils; (f) to stimulate DNA synthesis by human acute myelogenous leukemia (AML) cells; (g) to prime cells for production of leukotrienes and histamines; (h) to induce leukocyte chemotaxis; and (i) to induce cell surface molecules needed for leukocyte adhesion.

Mature human interleukin-3 (hIL-3) consists of 133 amino acids. It has one disulfide bridge and two potential glycosylation sites (Yang, et al., CELL 47:3 (1986)).

Murine IL-3 (mIL-3) was first identified by Ihle, et al., J. IMMUNOL. 126:2184 (1981) as a factor which induced expression of a T cell associated enzyme, 20-hydroxysteroid dehydrogenase. The factor was purified to homogeneity and shown to regulate the growth and differentiation of numerous subclasses of early hematopoietic and lymphoid progenitor cells.

In 1984, cDNA clones coding for murine IL-3 were isolated (Fung, et al., NATURE 307:233 (1984) and Yokota, et al., PROC. NATL. ACAD. SCI. USA 81:1070 (1984)). The murine DNA sequence coded for a polypeptide of 166 amino acids including a putative signal peptide.

The gibbon IL-3 sequence was obtained using a gibbon cDNA expression library. The gibbon IL-3 sequence was then used as a probe against a human genomic library to obtain a human IL-3 sequence.

Gibbon and human genomic DNA homologues of the murine IL-3 sequence were disclosed by Yang, et al., CELL 47:3 (1986). The human sequence reported by Yang, et al. included a serine residue at position 8 of the mature protein sequence. Following this finding, others reported isolation of Pro

8

hIL-3 cDNAs having proline at position 8 of the protein sequence. Thus it appears that there may be two allelic forms of hIL-3.

Dorssers, et al., GENE 55:115 (1987), found a clone from a human cDNA library which hybridized with mIL-3. This hybridization was the result of the high degree of homology between the 3′ noncoding regions of mIL-3 and hIL-3. This cDNA coded for an hIL-3 (Pro

8

) sequence.

U.S. Pat. No. 4,877,729 and U.S. Pat. No. 4,959,454 disclose human IL-3 and gibbon IL-3 cDNAs and the protein sequences for which they code. The hIL-3 disclosed has serine rather than proline at position 8 in the protein sequence.

Clark-Lewis, et al., SCIENCE 231:134 (1986) performed a functional analysis of murine IL-3 analogues synthesized with an automated peptide synthesizer. The authors concluded that the stable tertiary structure of the complete molecule was required for full activity. A study on the role of the disulfide bridges showed that replacement of all four cysteines by alanine gave a molecule with 1/500th the activity as the native molecule. Replacement of two of the four Cys residues by Ala(Cys

79

, Cys

140

→Ala

79

, Ala

140

) resulted in an increased activity. The authors concluded that in murine IL-3 a single disulfide bridge is required between cysteines 17 and 80 to get biological activity that approximates physiological levels and that this structure probably stabilizes the tertiary structure of the protein to give a conformation that is optimal for function. (Clark-Lewis, et al., PROC. NATL. ACAD. SCI. USA 85:7897 (1988)).

International Patent Application (PCT) WO 88/00598 discloses gibbon- and human-like IL-3. The hIL-3 contains a Ser

8

→Pro

8

replacement. Suggestions are made to replace Cys by Ser, thereby breaking the disulfide bridge, and to replace one or more amino acids at the glycosylation sites.

EP-A-0275598 (WO 88/04691) illustrates that Ala

1

can be deleted while retaining biological activity. Some mutant hIL-3 sequences are provided, e.g., two double mutants, Ala

1

→Asp

1

, Trp

13

→Arg

13

(pGB/IL-302) and Ala

1

→Asp

1

, Met

3

→Thr

3

(pGB/IL-304) and one triple mutant Ala

1

→Asp

1

, Leu

9

→Pro

9

, Trp

13

→Arg

13

(pGB/IL-303).

WO 88/05469 describes how deglycosylation mutants can be obtained and suggests mutants of Arg

54

Arg

55

and Arg

108

Arg

109

Lys

110

might avoid proteolysis upon expression in

Saccharomyces cerevisiae

by KEX2 protease. No mutated proteins are disclosed. Glycosylation and the KEX2 protease activity are only important, in this context, upon expression in yeast.

WO 88/06161 mentions various mutants which theoretically may be conformationally and antigenically neutral. The only actually performed mutations are Met

2

→Ile

2

and Ile

131

→Leu

131

. It is not disclosed whether the contemplated neutralities were obtained for these two mutations.

WO 91/00350 discloses nonglycosylated hIL-3 analog proteins, for example, hIL-3 (Pro

8

Asp

15

Asp

70

), Met

3

rhul-3 (Pro

8

Asp

15

Asp

70

); Thr

4

rhuL-3 (Pro

8

Asp

15

Asp

70

) and Thr

6

rhuIL-3 (Pro8Asp

15

Asp

70

). It is said that these protein compositions do not exhibit certain adverse side effects associated with native hIL-3 such as urticaria resulting from infiltration of mast cells and lymphocytes into the dermis. The disclosed analog hIL-3 proteins may have N termini at Met

3

, Thr

4

, or Thr

6

.

WO 91/12874 discloses cysteine added variants (CAVS) of IL-3 which have at least one Cys residue substituted for a naturally occurring amino acid residue.

SUMMARY OF THE INVENTION

The present invention relates to recombinant human interleukin-3 (hIL-3) variant or mutant proteins (muteins). These hIL-3 muteins contain amino acid substitutions and may also have amino acid deletions at either/or both the N- and C-termini. Preferably, these mutant polypeptides of the present invention contain four or more amino acids which differ from the amino acids found at the corresponding positions in the native hIL-3 polypeptide. The invention also relates to pharmaceutical compositions containing the hIL-3 muteins, DNA coding for the muteins, and methods for using the muteins. Additionally, the present invention relates to recombinant expression vectors comprising nucleotide sequences encoding the hIL-3 muteins, related microbial expression systems, and processes for making the hIL-3 muteins using the microbial expression systems.

The present invention includes mutants of hIL-3 in which from 1 to 14 amino acids have been deleted from the N-terminus and/or from 1 to 15 amino acids have been deleted from the C-terminus, and in which multiple amino acid substitutions have been made. Preferred muteins of the present invention are those in which amino acids 1 to 14 have been deleted from the N-terminus, amino acids 126 to 133 have been deleted from the C-terminus, and which also contain from about four to about twenty-six amino acid substitutions in the polypeptide sequence. These hIL-3 multiple mutation polypeptides may have biological activities similar to or better than hIL-3 and, in some cases, may also have an improved side effect profile, i.e., some muteins may have a better therapeutic index than native hIL-3. The present invention also provides muteins which may function as IL-3 antagonists or as discrete antigenic fragments for the production of antibodies useful in immunoassay and immunotherapy protocols. In addition to the use of the hIL-3 multiple mutation polypeptides of the present invention in vivo, it is envisioned that in vitro uses would include the ability to stimulate bone marrow and blood cell activation and growth before infusion into patients.

Antagonists of hIL-3 would be particularly useful in blocking the growth of certain cancer cells like AML, CML and certain types of B lymphoid cancers. Other conditions where antagonists would be useful include those in which certain blood cells are produced at abnormally high numbers or are being activated by endogenous ligands. Antagonists would effectively compete for ligands, presumably naturally occurring hemopoietins including and not limited to IL-3, GM-CSF and IL-5, which might trigger or augment the growth of cancer cells by virtue of their ability to bind to the IL-3 receptor complex while intrinsic activation properties of the ligand are diminished. IL-3, GM-CSF and/or IL-5 also play a role in certain asthmatic responses. An antagonist of the IL-3 receptor may have the utility in this disease by blocking receptor-mediated activation and recruitment of inflammatory cells.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1

is the human IL-3 gene for

E. coli

expression (pMON5873), encoding the polypeptide sequence of natural (wild type) human IL-3 [SEQ ID NO:128], plus an initiator methionine, as expressed in

E. coli,

with the amino acids numbered from the N-terminus of the natural hIL-3.

FIG.

2

: ClaI to NsiI Replacement Fragment.

FIG. 2

shows the nucleotide sequence of the replacement fragment used between the ClaI and NsiI sites of the hIL-3 gene. The codon choice used in the fragment corresponds to that found in highly expressed

E. coli

genes (Gouy and Gautier, 1982). Three new unique restriction sites, EcoRV, XhoI and PstI were introduced for the purpose of inserting synthetic gene fragments. The portion of the coding sequence shown encodes hIL-3 amino acids 20-70.

FIGS. 3A and 3B

shows the nucleotide and amino acid sequence of the gene in pMON5873 with the sequence extending from NcoI through HindIII. The codon choices used to encode amino acids 1-14 and 107-133 correspond to that found in highly expressed

E. coli

genes.

FIG. 4

shows the construction of the plasmid vector pMON5846 which encodes [Met-(1-133) hIL-3 (Arg

129

)].

FIG. 5

shows the construction of the plasmid vector pMON5847 (ATCC 68912) which encodes [Met-(1-133) hIL-3 (Arg

129

)].

FIG. 6

shows the construction of plasmid vector pMON5853 which encodes [Met-(15-133) hIL-3 (Arg

129

)].

FIG. 7

shows the construction of the plasmid vector pMON5854 which encodes [Met-(1-133) hIL-3 (Arg

129

)].

FIG. 8

shows the DNA sequence and resulting amino acid sequence of the LamB signal peptide.

FIG. 9

shows the construction of the plasmid vector pMON5978 which encodes Met-Ala-(15-125)hIL-3.

FIG. 10

shows the construction of the plasmid vector pMON5988 which encodes Met-Ala(15-125)hIL-3.

FIG. 11

shows the construction of the plasmid vector pMON5887 which encodes Met-(1-125)hIL-3.

FIG. 12

shows the construction of pMON6457 which encodes (15-125)hIL-3; it contains the araBAD promoter and the LamB signal peptide fused to the variant hIL-3 amino acids 15-125.

FIG. 13

shows the construction of pMON6458; it contains the araBAD promoter and the LamB signal peptide fused to the variant hIL-3 amino acids 15-125.

FIG. 14

shows the construction of pMON13359.

FIG. 15

shows the construction of pMON13352.

FIG. 16

shows the construction of pMON13360.

FIG. 17

shows the construction of pMON13363.

FIG. 18

shows the construction of pMON13364.

FIG. 19

shows the construction of pMON13365.

FIG. 20

shows the construction of pMON13287.

FIG. 21

shows the construction of pMON13288.

FIG. 22

shows the construction of pMON13289.

FIG. 23

shows the construction of pMON5723.

FIG. 24

shows the construction of pMON13438.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to muteins of human interleukin-3 (hIL-3) in which amino acid substitutions have been made at four or more positions in amino acid sequence of the polypeptide and to muteins which have substantially the same structure and substantially the same biological activity. Preferred muteins of the present invention are (15-125)hIL-3 deletion mutants which have deletions of amino acids 1 to 14 at the N-terminus and 126 to 133 at the C-terminus and which also have four or more amino acid substitutions in the polypeptide and muteins having substantially the same structure and substantially the same biological activity. Among the preferred muteins are those having twenty-six amino acid substitutions. As used herein human interleukin-3 corresponds to the amino acid sequence (1-133) as depicted in FIG.

1

and (15-125) hIL-3 corresponds to the 15 to 125 amino acid sequence of the hIL-3 polypeptide. Naturally occurring variants of hIL-3 polypeptide amino acids are also included in the present invention (for example, the allele in which proline rather than serine is at position 8 in the hIL-3 polypeptide sequence) as are variant hIL-3 molecules which are modified post-translationally (e.g. glycosylation).

The present invention also includes the DNA sequences which code for the mutant polypeptides, DNA sequences which are substantially similar and perform substantially the same function, and DNA sequences which differ from the DNAs encoding the muteins of the invention only due to the degeneracy of the genetic code.

Included in the present invention are novel mutant human interleukin-3 polypeptides comprising a polypeptide having the amino acid sequence of native human interleukin-3 wherein amino acids 126 to 133 have been deleted from the C-terminus of the native human interleukin-3 polypeptide and amino acids 1 to 14 have been deleted from the N-terminus of the native human interleukin-3 polypeptide and, in addition, polypeptides also have four or more amino acid substitutions in the polypeptide sequence.

Also included in the present invention are the DNA sequences coding for the muteins of the present invention; the oligonucleotide intermediates used to construct the mutant DNAS; and the polypeptides coded for by these oligonucleotides. These polypeptides may be useful as antagonists or as antigenic fragments for the production of antibodies useful in imnunoassay and immunotherapy protocols.

The mutant hIL-3 polypeptides of the present invention may also have methionine, alanine, or methionine-alanine residues inserted at the N-terminus.

The present invention includes human interleukin-3 mutant polypeptide Formula I:

Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn

1 5 10 15

Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa

35 40 45

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

50 55 60

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

65 70 75

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

80 85 90

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

95 100 105

Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

110 115 120

Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe [SEQ ID NO:15]

125 130

wherein Xaa at position 17 is Ser, Lys, Gly, Asp, Met, Gln, or

Arg;

Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

Xaa at position 19 is Met, Phe, Ile, Arg, Gly, Ala, or Cys;

Xaa at position 20 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;

Xaa at position 21 is Asp, Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn,

Thr, Ser or Val;

Xaa at position 22 is Glu, Trp, Pro, Ser, Ala, His, Asp, Asn, Gln,

Leu, Val or Gly;

Xaa at position 23 is Ile, Val1 Ala, Leu, Gly, Trp, Lys, Phe,

Ser, or Arg;

Xaa at position 24 is Ile, Gly, Val, Arg, Ser, Phe, or Leu;

Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

Xaa at position 26 is His, Thr, Phe, Gly, Arg, Ala, or Trp;

Xaa at position 27 is Leu, Gly, Arg, Thr, Ser, or Ala;

Xaa at position 28 is Lys, Arg, Leu, Gln, Gly, Pro, Val or Trp;

Xaa at position 29 is Gln, Asn, Leu, Pro, Arg, or Val;

Xaa at position 30 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu,

or Lys;

Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

Xaa at position 32 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or

Glu;

Xaa at position 33 is Pro, Leu, Gln, Ala, Thr, or Glu;

Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Glu, Gln, Thr,

Arg, Ala, Phe, Ile or Met;

Xaa at position 35 is Leu, Ala, Gly, Asn, Pro, Gln, or Val;

Xaa at position 36 is Asp, Leu, or Val;

Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile;

Xaa at position 38 is Asn, or Ala;

Xaa at position 40 is Leu, Trp, or Arg;

Xaa at position 41 is Asn, Cys, Arg, Leu, His, Met, or Pro;

Xaa at position 42 is Gly, Asp, Ser, Cys, Asn, Lys, Thr, Leu,

Val, Glu, Phe, Tyr, Ile, Met or Ala;

Xaa at position 43 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys,

Gln, Arg, Thr, Gly or Ser;

Xaa at position 44 is Asp, Ser, Leu, Arg, Lys, Thr, Met, Trp,

Glu, Asn, Gln, Ala or Pro;

Xaa at position 45 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys,

Trp, Asp, Asn, Arg, Ser, Ala, Ile, Glu or His;

Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Giu, Asn, Gln,

Lys, His, Ala, Tyr, Ile, Val or Gly;

Xaa at position 47 is Ile, Gly, Val, Ser, Arg, Pro, or His;

Xaa at position 48 is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu,

Lys, Thr, Ala, Met, Val or Asn;

Xaa at position 49 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;

Xaa at position 50 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser,

Ala, Ile, Val, His, Phe, Met or Gln;

Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;

Xaa at position 52 is Asn, His, Arg, Leu, Gly, Ser, or Thr;

Xaa at position 53 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser,

or Met;

Xaa at position 54 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn,

Lys, His, Ala or Leu;

Xaa at position 55 is Arg, Thr, Val, Ser, Leu, or Gly;

Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His,

Thr, Ala, Tyr, Phe, Leu, Val or Lys;

Xaa at position 57 is Asn or Gly;

Xaa at position 58 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;

Xaa at position 59 is Glu Tyr, His, Leu, Pro, or Arg;

Xaa at position 60 is Ala, Ser, Pro, Tyr, Asn, or Thr;

Xaa at position 61 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser;

Xaa at position 62 is Asn His, Val, Arg, Pro, Thr, Asp, or Ile;

Xaa at position 63 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or

Val;

Xaa at position 64 is Ala, Asn, Pro, Ser, or Lys;

Xaa at position 65 is Val, Thr, Pro, His, Leu, Phe, or Ser;

Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

Xaa at position 67 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro,

or His;

Xaa at position 68 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or

His;

Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly,

or Leu;

Xaa at position 70 is Asn, Leu, Val, Trp, Pro, or Ala;

Xaa at position 71 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln,

Trp, or Asn;

Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or

Asp;

Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or

Arg;

Xaa at position 74 is Ile, Met, Thr, Pro, Arg, Gly, Ala;

Xaa at position 75 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser,

Gln, or Leu;

Xaa at position 76 is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly,

or Asp;

Xaa at position 77 is Ile, Ser, Arg, Thr, or Leu;

Xaa at position 78 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg;

Xaa at position 79 is Lys, Thr, Asn, Met, Arg, Ile, Gly, or

Asp;

Xaa at position 80 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or

Arg;

Xaa at position 81 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or

Lys;

Xaa at position 82 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn,

His, Thr, Ser, Ala, Tyr, Phe, Ile, Met or Val;

Xaa at position 83 is Pro, Ala, Thr, Trp, Arg, or Met;

Xaa at position 84 is Cys, Glu, Gly, Arg, Met, or Val;

Xaa at position 85 is Leu, Asn, Val, or Gln;

Xaa at position 86 is Pro, Cys, Arg, Ala, or Lys;

Xaa at position 87 is Leu, Ser, Trp, or Gly;

Xaa at position 88 is Ala, Lys, Arg, Val, or Trp;

Xaa at position 89 is Thr, Asp, Cys, Leu, Val, Glu, His, Asn,

or Ser;

Xaa at position 90 is Ala, Pro, Ser, Thr, Gly, Asp, Ile, or

Met;

Xaa at position 91 is Ala, Pro, Ser, Thr, Phe, Leu, Asp, or His;

Xaa at position 92 is Pro, Phe, Arg, Ser, Lys, His, Ala, Gly, Ile

or Leu;

Xaa at position 93 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;

Xaa at position 94 is Arg, Ile, Ser, Glu, Leu, Val, Gln, Lys, His,

Ala,

or Pro;

Xaa at position 95 is His, Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn,

Lys,

Ser, Ala, Trp, Phe, Ile, or Tyr;

Xaa at position 96 is Pro, Lys, Tyr, Gly, Ile, or Thr;

Xaa at position 97 is Ile, Val, Lys, Ala, or Asn;

Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr,

Glu, Gln, Ser, Phe, Met, Val, Lys, Arg, Tyr or Pro;

Xaa at position 99 is Ile, Leu, Arg, Asp, Val, Pro, Gln,

Gly, Ser, Phe, or His;

Xaa at position 100 is Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln,

or Pro;

Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val,

Tyr, Glu, Asn, Ser, Ala, Gly, Ile, Leu, or Gln;

Xaa at position 102 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;

Xaa at position 103 is Asp, or Ser;

Xaa at position 104 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu,

Gln, Lys, Ala, Phe, or Gly;

Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,

Leu, Lys, Ile, Asp, or His;

Xaa at position 106 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;

Xaa at position 108 is Arg, Lys, Asp, Leu, Thr, Ile, Gln, His, Ser,

Ala

or Pro;

Xaa at position 109 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly;

Xaa at position 110 is Lys, Ala, Asn, Thr, Leu, Arg, Gln, His, Glu,

Ser,

or Trp;

Xaa at position 111 is Leu, Ile, Arg, Asp, or Met;

Xaa at position 112 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe;

Xaa at position 113 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp,

Lys, Leu, Ile, Val or Asn;

Xaa at position 114 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;

Xaa at position 115 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr,

Trp, or Met;

Xaa at position 116 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu,

Arg, Trp, Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile;

Xaa at position 117 is Thr, Ser, Asn, Ile, Trp, Lys, or Pro;

Xaa at position 118 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;

Xaa at position 119 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg;

Xaa at position 120 is Asn, Ala, Pro, Leu, His, Val, or Gln;

Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or

Gly;

Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,

Ile, Tyr, or Cys;

Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3.

Included in the present invention are human interleukin-3 mutant polypeptide of the Formula II:

Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn

1 5 10 15

Cys Xaa Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa Xaa

35 40 45

Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa

50 55 60

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

65 70 75

Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa Xaa

80 85 90

Xaa Xaa Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa

95 100 105

Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa Xaa

110 115 120

Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe [SEQ ID NO:16]

125 130

wherein

Xaa at position 17 is Ser, Gly, Asp, Met, or Gln;

Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

Xaa at position 19 is Met, Phe, Ile, Arg, or Ala;

Xaa at position 20 is Ile or Pro;

Xaa at position 21 is Asp or Glu;

Xaa at positicn 23 is Ile, Val, Ala, Leu, or Gly;

Xaa at position 24 is Ile, Val, Phe, or Leu;

Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

Xaa at position 26 is His, Phe, Gly, Arg, or Ala;

Xaa at position 28 is Lys, Leu, Gln, Gly, Pro, or Val;

Xaa at position 29 is Gln, Asn, Leu, Arg, or Val;

Xaa at position 30 is Pro, His, Thr, Gly, or Gln;

Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

Xaa at position 33 is Pro, Leu, Gln, Ala, or Glu;

Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu,

Ile, Phe, Thr or Met;

Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val;

Xaa at position 36 is Asp or Leu;

Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile;

Xaa at position 38 is Asn or Ala;

Xaa at position 41 is Asn, Cys, Arg, His, Met, or Pro;

Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met,

Tyr, Val or Arg;

Xaa at position 44 is Asp or Glu;

Xaa at position 45 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn, Glu,

Ser, or Trp;

Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu,

His, Ile, Lys, Tyr, Val or Gly;

Xaa at position 47 is Ile, Val, or His;

Xaa at position 49 is Met, Asn, or Asp;

Xaa at position 50 is Glu, Thr, Ala, Asn, Ser or Asp;

Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;

Xaa at position 52 is Asn or Gly;

Xaa at position 53 is Leu, Met, or Phe;

Xaa at position 54 is Arg, Ala, or Ser;

Xaa at position 55 is Arg, Thr, Val, Leu, or Gly;

Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Ala, Arg, Asn, Glu,

His,

Leu, Thr, Val or Lys;

Xaa at position 59 is Glu, Tyr, His, Leu, or Arg;

Xaa at position 60 is Ala, Ser, Asn, or Thr;

Xaa at position 61 is Phe or Ser;

Xaa at position 62 is Asn, Val, Pro, Thr, or Ile;

Xaa at position 63 is Arg, Tyr, Lys, Ser, His, or Val;

Xaa at position 64 is Ala or Asn;

Xaa at position 65 is Val, Thr, Leu, or Ser;

Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

Xaa at position 67 is Ser, Phe, Val, Gly, Asn, Ile, or His;

Xaa at position 68 is Leu, Val, Ile, Phe, or His;

Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

Xaa at position 70 is Asn or Pro;

Xaa at position 71 is Ala, Met, Pro, Arg, Glu, Thr, or Gln;

Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;

Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, Arg, or

Pro;

Xaa at position 74 is Ile or Met;

Xaa at position 75 is Glu, Gly, Asp, Ser, or Gln;

Xaa at position 76 is Ser, Val, Ala, Asn, Glu, Pro, Gly, or

Asp;

Xaa at position 77 is Ile, Ser, or Leu;

Xaa at position 79 is Lys, Thr, Asn, Met, Arg, Ile, Gly, or

Asp;

Xaa at position 80 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;

Xaa at position 81 is Leu, or Val;

Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu, His,

Met, Phe, Ser, Thr, Tyr or Val;

Xaa at position 83 is Pro, Ala, Thr, Trp, or Met;

Xaa at position 85 is Leu or Val;

Xaa at position 87 is Leu or Ser;

Xaa at position 88 is Ala, Arg, or Trp;

Xaa at position 89 is Thr, Asp, Glu, His, Asn, or Ser;

Xaa at position 90 is Ala, Asp, or Met;

Xaa at position 91 is Ala, Pro, Ser, Thr, Phe, Leu, or Asp;

Xaa at position 92 is Pro or Ser;

Xaa at position 93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

Xaa at position 95 is His, Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe,

Ser or Thr;

Xaa at position 96 is Pro or Tyr;

Xaa at position 97 is Ile, Val, or Ala;

Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr, Leu, Arg,

Gln,

Glu, lys, Met, Ser, Tyr, Val or Pro;

Xaa at position 99 is Ile, Leu, Val, or Phe;

Xaa at position 100 is Lys, Leu, His, Arg, Ile, Gln, Pro, or

Ser;

Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val,

Asn, Ile, Leu or Tyr;

Xaa at position 102 is Gly, Glu, Lys, or Ser;

Xaa at position 104 is Trp, Val, Tyr, Met, or Leu;

Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,

Leu, Lys, Ile, Asp, or His;

Xaa at position 106 is Glu, Ser, Ala, or Gly;

Xaa at position 108 is Arg, Ala, Gln, Ser or Lys;

Xaa at position 109 is Arg, Thr, Glu, Leu, Ser, or Gly;

Xaa at position 112 is Thr, Val, Gln, Glu, His, or Ser;

Xaa at position 114 is Tyr or Trp;

Xaa at position 115 is Leu or Ala;

Xaa at position 116 is Lys, Thr, Met, Val, Trp, Ser, Leu, Ala,

Asn,

Gln, His, Met, Phe, Tyr or Ile;

Xaa at position 117 is Thr, Ser, or Asn;

Xaa at position 119 is Glu, Ser, Pro, Leu, Thr, or Tyr;

Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;

Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or

Gly;

Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,

Ile, Tyr, or Cys;

Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn

1 5 10 15

Cys Xaa Xaa Xaa Ile Xaa Glu Xaa Xaa Xaa Xaa Leu Lys Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa Xaa

35 40 45

Xaa Ile Leu Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Glu Xaa

50 55 60

Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile Glu

65 70 75

Xaa Xaa Leu Xaa Xaa Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr Ala

80 85 90

Xaa Pro Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa Xaa

95 100 105

Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Glu Xaa

110 115 120

Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe [SEQ ID NO:17]

125 130

wherein

Xaa at position 17 is Ser, Gly, Asp, Met, or Gln;

Xaa at position 18 is Asn, His, or Ile;

Xaa at position 19 is Met or Ile;

Xaa at position 21 is Asp or Glu;

Xaa at position 23 is Ile, Ala, Leu, or Gly;

Xaa at position 24 is Ile, Val, or Leu;

Xaa at position 25 is Thr, His, Gln, or Ala;

Xaa at position 26 is His or Ala;

Xaa at position 29 is Gln, Asn, or Val;

Xaa at position 30 is Pro, Gly, or Gln;

Xaa at position 31 is Pro, Asp, Gly, or Gln;

Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

Xaa at position 33 is Pro or Glu;

Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln,

Glu, Ile, Phe, Thr or Met;

Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val;

Xaa at position 37 is Phe, Ser, Pro, or Trp;

Xaa at position 38 is Asn or Ala;

Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu,

Met, Tyr or Arg;

Xaa at position 44 is Asp or Glu;

Xaa at position 45 is Gln, Val, Met, Leu, Thr, Ala, Asn, Glu,

Ser or Lys;

Xaa at position 46 is Asp, Phe, Ser, Thr, Ala, Asn Gln, Glu, His,

Ile, Lys, Tyr, Val or Cys;

Xaa at position 50 is Glu, Ala, Asn, Ser or Asp;

Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;

Xaa at position 54 is Arg or Ala;

Xaa at position 54 is Arg or Ala;

Xaa at position 55 is Arg, Thr, Val, Leu, or Gly;

Xaa at position 56 is Pro, Gly, Ser, Gln, Ala, Arg, Asn, Glu,

Leu, Thr, Val or Lys;

Xaa at position 60 is Ala or Ser;

Xaa at position 62 is Asn, Pro, Thr, or Ile;

Xaa at position 63 is Arg or Lys;

Xaa at position 64 is Ala or Asn;

Xaa at position 65 is Val or Thr;

Xaa at position 66 is Lys or Arg;

Xaa at position 67 is Ser, Phe, or His;

Xaa at position 68 is Leu, Ile, Phe, or His;

Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

Xaa at position 71 is Ala, Pro, or Arg;

Xaa at position 72 is Ser, Glu, Arg, or Asp;

Xaa at position 73 is Ala or Leu;

Xaa at position 76 is Ser, Val, Ala, Asn, Glu, Pro, or Gly;

Xaa at position 77 is Ile or Leu;

Xaa at position 79 is Lys, Thr, Asn, Met, Arg, Ile, Gly, or

Asp;

Xaa at position 80 is Asn, Gly, Glu, or Arg;

Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu, His,

Ile, Met, Phe, Ser, Thr, Tyr or Val;

Xaa at position 83 is Pro or Thr;

Xaa at position 85 is Leu or Val;

Xaa at position 87 is Leu or Ser;

Xaa at position 88 is Ala or Trp;

Xaa at position 91 is Ala or Pro;

Xaa at position 93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

Xaa at position 95 is His, Pro, Arg, Val, Leu1, Gly, Asn, Phe, Ser

or Thr;

Xaa at position 96 is Pro or Tyr;

Xaa at position 97 is Ile or Val;

Xaa at position 98 is His, Ile, Asn, Leu, Ala, Thr, Arg, Gln,

Lys, Met, Ser, Tyr, Val or Pro;

Xaa at position 99 is Ile, Leu, or Val;

Xaa at position 100 is Lys, Arg, Ile, Gln, Pro, or Ser;

Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Pro, Asn,

Ile, Leu or Tyr;

Xaa at position 104 is Trp or Leu;

Xaa at position 105 is Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu,

Lys, Ile, Asp, or His;

Xaa at position 106 is Glu or Gly;

Xaa at position 108 is Arg, Ala, or Ser;

Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser;

Xaa at position 112 is Thr, Val, or Gln;

Xaa at position 114 is Tyr or Trp;

Xaa at position 115 is Leu or Ala;

Xaa at position 116 is Lys, Thr, Val, Trp, Ser, Ala, His, Met,

Phe, Tyr or Ile;

Xaa at position 117 is Thr or Ser;

Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;

Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Asp, or Gly;

Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,

Ile, Tyr, or Cys;

Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 35 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133)human interleukin-3.

Included in the present invention are human interleukin-3 mutant polypeptide of the Formula IV:

Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn

1 5 10 15

Cys Xaa Xaa Met Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa Xaa

20 25 30

Pro Xaa Pro Xaa Xaa ASP Phe Xaa Asn Leu Asn Xaa Glu Asp Xaa

35 40 45

Xaa Ile Leu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Glu Ala

50 55 60

Phe Xaa Arg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile Glu

65 70 75

Xaa Xaa Leu Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr Ala

80 85 90

Xaa Pro Xaa Arg Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp Xaa

95 100 105

Glu Phe Xaa Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu Xaa

110 115 120

Xaa Xaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe [SEQ ID NO:18)

125 130

wherein

Xaa at position 17 is Ser, Gly, Asp, or Gln;

Xaa at position 18 is Asn, His, or Ile;

Xaa at position 23 is Ile, Ala, Leu, or Gly;

Xaa at position 25 is Thr, His, or Gln;

Xaa at position 26 is His or Ala;

Xaa at position 29 is Gln or Asn;

Xaa at position 30 is Pro or Gly;

Xaa at position 32 is Leu, Arg, Asn, or Ala;

Xaa at position 34 is Leu, Val, Ser, Ala, Arg, Gln, Glu, Ile,

Phe, Thr, or Met;

Xaa at position 35 is Leu, Ala, Asn, or Pro;

Xaa at position 38 is Asn or Ala;

Xaa at position 42 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met,

Tyr or Arg;

Xaa at position 45 is Gln, Val, Met, Leu, Ala, Asn, Glu, or Lys;

Xaa at position 46 is Asp, Phe, Ser, Gln, Glu, His, Val

or Thr;

Xaa at position 50 is Glu Asn, Ser or Asp;

Xaa at position 51 is Asn, Arg, Pro, Thr, or His;

Xaa at position 55 is Arg, Leu, or Gly;

Xaa at position 56 is Pro, Gly, Ser, Ala, Asn, Val, Leu or Gln;

Xaa at position 62 is Asn, Pro, or Thr;

Xaa at position 64 is Ala or Asn;

Xaa at position 65 is Val or Thr;

Xaa at position 67 is Ser or Phe;

Xaa at position 68 is Leu or Phe;

Xaa at position 69 is Gln, Ala, Glu, or Arg;

Xaa at position 76 is Ser, Val, Asn, Pro, or Gly;

Xaa at position 77 is Ile or Leu;

Xaa at position 79 is Lys, Asn, Met, Arg, Ile, or Gly;

Xaa at position 80 is Asn, Gly, Glu, or Arg;

Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His, Met,

Phe, Ser, Thr, Tyr or Val;

Xaa at position 87 is Leu or Ser;

Xaa at position 88 is Ala or Trp;

Xaa at position 91 is Ala or Pro;

Xaa at position 93 is Thr, Asp, or Ala;

Xaa at position 95 is His, Pro, Arg, Val, Gly, Asn, Ser or Thr;

Xaa at position 98 is His, Ile, Asn, Ala, Thr, Gln, Glu,

Lys, Met, Ser, Tyr, Val or Leu;

Xaa at position 99 is Ile or Leu;

Xaa at position 100 is Lys or Arg;

Xaa at position 101 is Asp, Pro, Met, Lys, Thr, His, Pro, Asn, Ile,

Leu or Tyr;

Xaa at position 105 is Asn, Pro, Ser, Ile or Asp;

Xaa at position 108 is Arg, Ala, or Ser;

Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser;

Xaa at position 112 is Thr or Gln;

Xaa at position 116 is Lys, Val, Trp, Ala, His, Phe, Tyr or Ile;

Xaa at position 117 is Thr or Ser;

Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;

Xaa at position 121 is Ala, Ser, Ile, Pro, or Asp;

Xaa at position 122 is Gln, Met, Trp, Phe, Pro, His, Ile, or Tyr;

Xaa at position 123 is Ala, Met, Glu, Ser, or Leu;

and which can additionally have Met- preceding the amino acid in position 1; and wherein from 1 to 14 amino acids can be deleted from the N-terminus and/or from 1 to 15 amino acids can be deleted from the C-terminus; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133)human interleukin-3.

Included in the present invention are (15-125)human interleukin-3 mutant polypeptides of the Formula V:

Asn Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

1 5 10 15

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa

20 25 30

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

35 40 45

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

50 55 60

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

65 70 75

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

80 85 90

Xaa Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

95 100 105

Xaa Xaa Xaa Xaa Gln Gln [SEQ ID NO:19]

110

wherein

Xaa at position 3 is Ser, Lys, Gly, Asp, Met, Gln, or Arg;

Xaa at position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

Xaa at position 5 is Met, Phe, Ile, Arg, Gly, Ala, or Cys;

Xaa at position 6 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;

Xaa at position 7 is Asp, Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn,

Thr, Ser or Val;

Xaa at position 8 is Glu, Trp, Pro, Ser, Ala, His, Asp, Asn, Gln,

Leu, Val, or Gly;

Xaa at position 9 is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe,

Ser, or Arg;

Xaa at position 10 is Ile, Gly, Val, Arg, Ser, Phe, or Leu;

Xaa at position 11 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

Xaa at position 12 is His, Thr, Phe, Gly, Arg, Ala, or Trp;

Xaa at position 13 is Leu, Gly, Arg, Thr, Ser, or Ala;

Xaa at position 14 is Lys, Arg, Leu, Gln, Gly, Pro, Val or Trp;

Xaa at position 15 is Gln, Asn, Leu, Pro, Arg, or Val;

Xaa at position 16 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, or

Lys;

Xaa at position 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

Xaa at position 18 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu;

Xaa at position 19 is Pro, Leu, Gln, Ala, Thr, or Glu;

Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Glu, Gln, Thr,

Arg, Ala, Phe, Ile or Met;

Xaa at position 21 is Leu, Ala, Gly, Asn, Pro, Gln, or Val;

Xaa at position 22 is Asp, Leu, or Val;

Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile;

Xaa at position 24 is Asn, or Ala;

Xaa at position 26 is Leu, Trp, or Arg;

Xaa at position 27 is Asn, Cys, Arg, Leu, His, Met, Pro;

Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Lys, Asn, Thr, Leu,

Val, Glu, Phe, Tyr, Ile or Met;

Xaa at position 29 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys, Gln,

Arg, Thr, Gly or Ser;

Xaa at position 30 is Asp, Ser, Leu, Arg, Lys, Thr,Met, Trp, Glu,

Asn, Gln, Ala or Pro;

Xaa at position 31 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys, Asp,

Asn, Arg, Ser, Ala, Ile, Glu, His or Trp;

Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Glu, Asn, Gln,

Lys, His, Ala, Tyr, Ile, Val or Gly;

Xaa at position 33 is Ile, Gly, Val, Ser, Arg, Pro, or His;

Xaa at position 34 is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu,

Lys, Thr, Ala, Met, Val or Asn;

Xaa at position 35 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;

Xaa at position 36 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala,

Ile, Val, His, Phe, Met or Gln;

Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;

Xaa at position 38 is Asn, His, Arg, Leu, Gly, Ser, or Thr;

Xaa at position 39 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser,

Met, or;

Xaa at position 40 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn,

Lys, His, Ala or Leu:

Xaa at position 41 is Arg, Thr, Val, Ser, Leu, or Gly;

Xaa at position 42 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His,

Thr, Ala, Tyr, Phe, Leu, Val or Lys;

Xaa at position 43 is Asn or Gly;

Xaa at position 44 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;

Xaa at position 45 is Glu Tyr, His, Leu, Pro, or Arg:

Xaa at position 46 is Ala, Ser, Pro, Tyr, Asn, or Thr;

Xaa at position 47 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser;

Xaa at position 48 is Asn, His, Val, Arg, Pro, Thr, Asp, or Ile;

Xaa at position 49 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or Val;

Xaa at position 50 is Ala, Asn, Pro, Ser, or Lys;

Xaa at position 51 is Val, Thr, Pro, His, Leu, Phe, or Ser;

Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

Xaa at position 53 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, or

His;

Xaa at position 54 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or His;

Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly, or

Leu;

Xaa at position 56 is Asn, Leu, Val, Trp, Pro, or Ala;

Xaa at position 57 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln,

Trp, or Asn;

Xaa at position 58 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;

Xaa at position 59 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or Arg;

Xaa at position 60 is Ile, Met, Thr, Pro, Arg, Gly, Ala;

Xaa at position 61 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser,

Gln, or Leu;

Xaa at position 62 is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly, or

Asp;

Xaa at position 63 is Ile, Ser, Arg, Thr, or Leu;

Xaa at position 64 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg;

Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or

Asp;

Xaa at position 66 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or Arg;

Xaa at position 67 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys;

Xaa at position 68 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn,

His, Thr, Ser, Ala, Tyr, Phe, Ile, Met or Val;

Xaa at position 69 is Pro, Ala, Thr, Trp, Arg, or Met;

Xaa at position 70 is Cys, Glu, Gly, Arg, Met, or Val;

Xaa at position 71 is Leu, Asn, Val, or Gln;

Xaa at position 72 is Pro, Cys, Arg, Ala, or Lys;

Xaa at position 73 is Leu, Ser, Trp, or Gly;

Xaa at position 74 is Ala, Lys, Arg, Val, or Trp;

Xaa at position 75 is Thr, Asp, Cys, Leu, Val, Glu, His, Asn, or

Ser;

Xaa at position 76 is Ala, Pro, Ser, Thr, Gly, Asp, Ile, or Met;

Xaa at position 77 is Ala, Pro, Ser, Thr, Phe, Leu, Asp, or His;

Xaa at position 78 is Pro, Phe, Arg, Ser, Lys, His, Ala, Gly, Ile

or Leu:

Xaa at position 79 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;

Xaa at position 80 is Arg, Ile, Ser, Glu, Leu, Val, Gln, Lys, His,

Ala or Pro;

Xaa at position 81 is His, Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn,

Lys, Ser, Ala, Trp, Phe, Ile or Tyr;

Xaa at position 82 is Pro, Lys, Tyr, Gly, Ile, or Thr;

Xaa at position 83 is Ile, Val, Lys, Ala, or Asn;

Xaa at position 84 is His, Ile, Asn, Leu, Asp, Ala, Thr, Glu,

Gln, Ser, Phe, Met, Val, Lys, Arg, Tyr or Pro;

Xaa at position 85 is Ile, Leu, Arg, Asp, Val, Pro, Gln,

Gly, Ser, Phe, or His;

Xaa at position 86 is Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln,

Pro;

Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Val,

Tyr, Glu, Asn, Ser, Ala, Gly, Ile, Leu or Gln;

Xaa at position 88 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;

Xaa at position 89 is Asp, or Ser;

Xaa at position 90 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu,

Gln, Lys, Ala, Phe, or Gly;

Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,

Leu, Lys, Ile, Asp, or His;

Xaa at position 92 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;

Xaa at position 94 is Arg, Lys, Asp, Leu, Thr, Ile, Gln,

His, Ser, Ala, or Pro;

Xaa at position 95 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly;

Xaa at position 96 is Lys, Asn, Thr, Leu, Gln, Arg,

His, Glu, Ser, Ala or Trp;

Xaa at position 97 is Leu, Ile, Arg, Asp, or Met;

Xaa at position 98 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe;

Xaa at position 99 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp,

Lys, Leu, Ile, Val or Asn;

Xaa at position 100 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;

Xaa at position 101 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr,

Trp, or Met;

Xaa at position 102 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu, Arg,

Trp,

Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile;

Xaa at position 103 is Thr, Ser, Asn, Ile, Trp, Lys, or Pro;

Xaa at position 104 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;

Xaa at position 105 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg;

Xaa at position 106 is Asn, Ala, Pro, Leu, His, Val, or Gln;

Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or

Gly;

Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,

Ile, Tyr, or Cys;

Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

and which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding native amino acids of (1-133) human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.

Included in the present invention are (15-125)human interleukin-3 mutant polypeptides of the Formula VI:

Asn Cys Xaa Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa

1 5 10 15

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa

20 25 30

Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa

35 40 45

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

50 55 60

Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa

65 70 75

Xaa Xaa Xaa Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa

80 85 90

Xaa Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa

95 100 105

Xaa Xaa Xaa Xaa Gln Gln [SEQ ID NO:20]

110

wherein

Xaa at position 3 is Ser, Gly, Asp, Met, or Gln;

Xaa at position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

Xaa at position 5 is Met, Phe, Ile, Arg, or Ala;

Xaa at position 6 is Ile or Pro;

Xaa at position 7 is Asp, or Glu;

Xaa at position 9 is Ile, Val, Ala, Leu, or Gly;

Xaa at position 10 is Ile, Val, Phe, or Leu;

Xaa at position 11 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

Xaa at position 12 is His, Phe, Gly, Arg, or Ala;

Xaa at position 14 is Lys, Leu, Gln, Gly, Pro, or Val;

Xaa at position 15 is Gln, Asn, Leu, Arg, or Val;

Xaa at position 16 is Pro, His, Thr, Gly, or Gln;

Xaa at position 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

Xaa at position 19 is Pro, Leu, Gln, Ala, or Glu;

Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln,

Glu, Ile, Phe, Thr or Met;

Xaa at position 21 is Leu, Ala, Asn, Pro, Gln, or Val;

Xaa at position 22 is Asp or Leu;

Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile;

Xaa at position 24 is Asn or Ala;

Xaa at position 27 is Asn, Cys, Arg, His, Met, or Pro;

Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu,

Met, Tyr, or Arg;

Xaa at position 30 is Asp, or Glu;

Xaa at position 31 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn Glu,

Ser or Trp;

Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln,

Glu, His, Ile, Lys, Tyr, Val or Gly;

Xaa at position 33 is Ile, Val, or His;

Xaa at position 35 is Met, Asn, or Asp;

Xaa at position 36 is Glu, Thr, Ala, Asn, Ser or Asp;

Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;

Xaa at position 38 is Asn or Gly;

Xaa at position 39 is Leu, Met, or Phe;

Xaa at position 40 is Arg, Ala or Ser;

Xaa at position 41 is Arg, Thr, Val, Leu, or Gly;

Xaa at position 42 is Pro, Gly, Cys, Ser, Gln, Ala, Arg, Asn,

Glu, His, Leu, Thr, Val or Lys;

Xaa at position 45 is Glu, Tyr, His, Leu, or Arg;

Xaa at position 46 is Ala, Ser, Asn, or Thr;

Xaa at position 47 is Phe or Ser;

Xaa at position 48 is Asn, Val, Pro, Thr, or Ile;

Xaa at position 49 is Arg, Tyr, Lys, Ser, His, or Val;

Xaa at position 50 is Ala or Asn;

Xaa at position 51 is Val, Thr, Leu, or Ser;

Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

Xaa at position 53 is Ser, Phe, Val, Gly, Asn, Ile, or His;

Xaa at position 54 is Leu, Val, Ile, Phe, or His;

Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

Xaa at position 56 is Asn or Pro;

Xaa at position 57 is Ala, Met, Pro, Arg, Glu, Thr, or Gln;

Xaa at position 58 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp:

Xaa at position 59 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, Arg, or

Pro;

Xaa at position 60 is Ile or Met;

Xaa at position 61 is Glu, Gly, Asp, Ser, or Gln;

Xaa at position 62 is Ser, Val, Ala, Asn, Glu, Pro, Gly, or

Asp;

Xaa at position 63 is Ile, Ser, or Leu;

Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or

Asp;

Xaa at position 66 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;

Xaa at position 67 is Leu, or Val;

Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu,

His, Met, Phe, Ser, Thr, Tyr or Val;

Xaa at position 69 is Pro, Ala, Thr, Trp, or Met;

Xaa at position 71 is Leu or Val;

Xaa at position 73 is Leu or Ser;

Xaa at position 74 is Ala, Arg, or Trp;

Xaa at position 75 is Thr, Asp, Glu, His, Asn, or Ser;

Xaa at position 76 is Ala, Asp, or Met;

Xaa at position 77 is Ala, Pro, Ser, Thr, Phe, Leu, or Asp;

Xaa at position 78 is Pro or Ser:

Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe,

Ser or Thr;

Xaa at position 82 is Pro or Tyr;

Xaa at position 83 is Ile, Val, or Ala;

Xaa at position 84 is His, Ile, Asn, Leu, Asp, Ala, Thr,

Arg, Gln, Glu, Lys, Met, Ser, Tyr, Val or Pro;

Xaa at position 85 is Ile, Leu, Val, or Phe;

Xaa at position 86 is Lys, Leu, His, Arg, Ile, Gln, Pro or

Ser;

Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Val,

Asn, Ile, Leu or Tyr;

Xaa at position 88 is Gly, Glu, Lys, or Ser;

Xaa at position 90 is Trp, Val, Tyr, Met, or Leu;

Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,

Leu, Lys, Ile, Asp, or His:

Xaa at position 92 is Glu, Ser, Ala, or Gly;

Xaa at position 94 is Arg, Ala, Gln, Ser or Lys;

Xaa at position 95 is Arg, Thr, Glu, Leu, Ser, or Gly;

Xaa at position 98 is Thr, Val, Gln, Glu, His, or Ser;

Xaa at position 100 is Tyr or Trp;

Xaa at position 101 is Leu or Ala;

Xaa at position 102 is Lys, Thr, Met, Val, Trp, Ser, Leu,

Ala, Asn, Gln, His, Met, Phe, Tyr or Ile;

Xaa at position 103 is Thr, Ser, or Asn;

Xaa at position 105 is Glu, Ser, Pro, Leu, Thr, or Tyr;

Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;

Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or

Gly;

Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,

Ile, Tyr, or Cys;

Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu:

and which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 44 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133) human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.

Included in the present invention are (15-125)human interleukin-3 mutant polypeptides of the Formula VII:

Asn Cys Xaa Xaa Xaa Ile Xaa Glu Xaa Xaa Xaa Xaa Leu Lys Xaa

1 5 10 15

Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa

20 25 30

Xaa Xaa Ile Leu Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Glu

35 40 45

Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile

50 55 60

Glu Xaa Xaa Leu Xaa Xaa Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr

65 70 75

Ala Xaa Pro Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa

80 85 90

Xaa Xaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Glu

95 100 105

Xaa Xaa Xaa Xaa Gln Gln [SEQ ID NO:21]

110

wherein

Xaa at position 3 is Ser, Gly, Asp, Met, or Gln;

Xaa at position 4 is Asn, His, or Ile;

Xaa at position 5 is Met or Ile;

Xaa at position 7 is Asp or Glu;

Xaa at position 9 is Ile, Ala, Leu, or Gly;

Xaa at position 10 is Ile, Val, or Leu;

Xaa at position 11 is Thr, His, Gln, or Ala;

Xaa at position 12 is His or Ala;

Xaa at position 15 is Gln, Asn, or Val;

Xaa at position 16 is Pro, Gly, or Gln;

Xaa at position 17 is Pro, Asp, Gly, or Gln;

Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

Xaa at position 19 is Pro or Glu;

Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Ala, Arg,

Gln, Glu, Ile, Phe, Thr or Met;

Xaa at position 21 is Leu, Ala, Asn, Pro, Gln, or Val;

Xaa at position 23 is Phe, Ser, Pro, or Trp;

Xaa at position 24 is Asn or Ala;

Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile,

Leu, Met Tyr or Arg;

Xaa at position 30 is Asp or Glu;

Xaa at position 31 is Gln, Val, Met, Leu, Thr, Ala, Asn,

Glu, Ser or Lys;

Xaa at position 32 is Asp, Phe, Ser, Thr, Ala, Asn, Gln, Glu,

His, Ile, Lys, Tyr, Val or Cys;

Xaa at position 36 is Glu, Ala, Asn, Ser or Asp;

Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;

Xaa at position 40 is Arg or Ala;

Xaa at position 41 is Arg, Thr, Val, Leu, or Gly;

Xaa at position 42 is Pro, Gly, Ser, Gln, Ala, Arg, Asn,

Glu, Leu, Thr, Val or Lys;

Xaa at position 46 is Ala or Ser;

Xaa at position 48 is Asn, Pro, Thr, or Ile;

Xaa at position 49 is Arg or Lys;

Xaa at position 50 is Ala or Asn;

Xaa at position 51 is Val or Thr;

Xaa at position 52 is Lys or Arg;

Xaa at position 53 is Ser, Phe, or His;

Xaa at position 54 is Leu, Ile, Phe, or His;

Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

Xaa at position 57 is Ala, Pro, or Arg;

Xaa at position 58 is Ser, Glu, Arg, or Asp;

Xaa at position 59 is Ala or Leu;

Xaa at position 62 is Ser, Val, Ala, Asn, Glu, Pro, or Gly;

Xaa at position 63 is Ile or Leu;

Xaa at position 65 is Lys, Thr, Asn, Met, Arg, Ile, Gly,

or Asp;

Xaa at position 66 is Asn, Gly, Glu, or Arg;

Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu,

His, Ile, Met, Phe, Ser, Thr, Tyr or Val;

Xaa at position 69 is Pro or Thr;

Xaa at position 71 is Leu or Val:

Xaa at position 73 is Leu or Ser;

Xaa at position 74 is Ala or Trp;

Xaa at position 77 is Ala or Pro;

Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Phe,

Ser or Thr;

Xaa at position 82 is Pro or Tyr;

Xaa at position 83 is Ile or Val;

Xaa at position 84 is His, Ile, Asn, Leu, Ala, Thr, Arg,

Gln, Lys, Met, Ser, Tyr, Val or Pro;

Xaa at position 85 is Ile, Leu, or Val;

Xaa at position 86 is Lys, Arg, Ile, Gln, Pro, or Ser;

Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Asn, Ile,

Leu or Tyr;

Xaa at position 90 is Trp or Leu;

Xaa at position 91 is Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu,

Lys, Ile, Asp, or His;

Xaa at position 92 is Glu, or Gly;

Xaa at position 94 is Arg, Ala, or Ser;

Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser;

Xaa at position 98 is Thr, Val, or Gln;

Xaa at position 100 is Tyr or Trp;

Xaa at position 101 is Leu or Ala;

Xaa at position 102 is Lys, Thr, Val, Trp, Ser, Ala, His,

Met, Phe, Tyr or Ile;

Xaa at position 103 is Thr or Ser;

Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;

Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Asp, or Gly;

Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,

Ile, Tyr, or Cys;

Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 35 of the amino acids designated by Xaa are different from the corresponding amino acids of native human interleukin-3.

Included in the present invention are (15-125)human interleukin-3 mutant polypeptides of the Formula VIII:

Asn Cys Xaa Xaa Met Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa

1 5 10 15

Xaa Pro Xaa Pro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Glu Asp

20 25 30

Xaa Xaa Ile Leu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Glu

35 40 45

Ala Phe Xaa Arg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile

50 55 60

Glu Xaa Xaa Leu Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr

65 70 75

Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp

80 85 90

Xaa Glu Phe Xaa Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu

95 100 105

Xaa Xaa Xaa Xaa Gln Gln [SEQ ID NO:22]

110

wherein

Xaa at position 3 is Ser, Gly, Asp, or Gln;

Xaa at position 4 is Asn, His, or Ile;

Xaa at position 9 is Ile, Ala, Leu, or Gly;

Xaa at position 11 is Thr, His, or Gln;

Xaa at position 12 is His or Ala;

Xaa at position 15 is Gln or Asn;

Xaa at position 16 is Pro or Gly;

Xaa at position 18 is Leu, Arg, Asn, or Ala:

Xaa at position 20 is Leu, Val, Ser, Ala, Arg, Gln, Glu, Ile,

Phe, Thr or Met:

Xaa at position 21 is Leu, Ala, Asn, or Pro;

Xaa at position 24 is Asn or Ala;

Xaa at position 28 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met,

Tyr or Arg;

Xaa at position 31 is Gln, Val, Met, Leu, Ala, Asn, Glu or Lys;

Xaa at position 32 is Asp, Phe, Ser, Ala, Gln, Glu, His, Val

or Thr;

Xaa at position 36 is Glu, Asn, Ser or Asp;

Xaa at position 37 is Asn, Arg, Pro, Thr, or His;

Xaa at position 41 is Arg, Leu, or Gly;

Xaa at position 42 is Pro, Gly, Ser, Ala, Asn, Val, Leu or Gln;

Xaa at position 48 is Asn, Pro, or Thr;

Xaa at position 50 is Ala or Asn;

Xaa at position 51 is Val or Thr;

I

Xaa at position 53 is Ser or Phe;

Xaa at position 54 is Leu or Phe;

Xaa at position 55 is Gln, Ala, Glu, or Arg;

Xaa at position 62 is Ser, Val, Asn, Pro, or Gly;

Xaa at position 63 is Ile or Leu;

Xaa at position 65 is Lys, Asn, Met, Arg, Ile, or Gly;

Xaa at position 66 is Asn, Gly, Glu, or Arg;

Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His,

Met, Phe, Ser, Thr, Tyr or Val;

Xaa at position 73 is Leu or Ser;

Xaa at position 74 is Ala or Trp:

Xaa at position 77 is Ala or Pro;

Xaa at position 79 is Thr, Asp, or Ala;

Xaa at position 81 is His, Pro, Arg, Val, Gly, Asn, Ser or Thr;

Xaa at position 84 is His, Ile, Asn, Ala, Thr, Arg, Gln, Glu,

Lys, Met, Ser, Tyr, Val or Leu;

Xaa at position 85 is Ile or Leu;

Xaa at position 86 is Lya or Arg:

Xaa at position 87 is Asp, Pro, Met, Lys, His, Pro, Asn, Ile,

Leu or Tyr;

Xaa at position 91 is Asn, Pro, Ser, Ile or Asp;

Xaa at position 94 is Arg, Ala, or Ser;

Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser;

Xaa at position 98 is Thr or Gln;

Xaa at position 102 is Lys, Val, Trp, or Ile;

Xaa at position 103 is Thr, Ala, His, Phe, Tyr or Ser;

Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;

Xaa at position 107 is Ala, Ser, Ile, Pro, or Asp;

Xaa at position 108 is Gln, Met, Trp, Phe, Pro, His, Ile, or

Tyr;

Xaa at position 109 is Ala, Met, Glu, Ser, or Leu;

and which can additionally have Met- or Met-Ala- preceding the amino acid in position 1; and wherein from 4 to 26 of the amino acids designated by Xaa are different from the corresponding amino acids of native (1-133)human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.

The present invention includes polypeptides of the formula

1 5 10

[SEQ ID NO:129]

(Met)

m

-Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr

15 20

Ser Trp Val Asn Cys Ser Xaa Xaa Xaa Asp Glu Xaa Ile

25 30 35

Xaa His Leu Lys Xaa Pro Pro Xaa Pro Xaa Leu Asp Xaa

40 45 50

Xaa Asn Leu Asn Xaa Glu Asp Xaa Xaa Ile Leu Xaa Xaa

55 60

Xaa Asn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa Xaa

65 70 75

Ala Xaa Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa

80 85

Ile Leu Xaa Asn Leu Xaa Pro Cys Xaa Pro Xaa Xaa Thr

90 95 100

Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Ile Xaa Xaa Gly

105 110 115

Asp Trp Xaa Glu Phe Arg Xaa Lys Leu Xaa Phe Tyr Leu

120 125

Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln Thr Thr Leu

130

Ser Leu Ala Ile Phe

wherein m is 0 or 1; Xaa at position 18 is Asn or Ile; Xaa at position 19 is Met, Ala or Ile; Xaa at position 20 is Ile, Pro or Leu; Xaa at position 23 is Ile, Ala or Leu; Xaa at position 25 is Thr or His; Xaa at position 29 is Gln, Arg, Val or Leu; Xaa at position 32 is Leu, Ala, Asn or Arg; Xaa at position 34 is Leu or Ser; Xaa at position 37 is Phe, Pro, or Ser; Xaa at position 38 is Asn or Ala; Xaa at position 42 is Gly, Ala, Ser, Asp or Asn; Xaa at position 45 is Gln, Val, or Met; Xaa at position 46 is Asp or Ser; Xaa at position 49 is Met, Ile, Leu or Asp; Xaa at position 50 is Glu or Asp; Xaa at position 51 is Asn Arg or Ser; Xaa at position 55 is Arg, Leu, or Thr; Xaa at position 56 is Pro or Ser; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaa at position 62 is Asn, Val or Pro; Xaa at position 63 is Arg or His; Xaa at position 65 is Val or Ser; Xaa at position 67 is Ser, Asn, His or Gly; Xaa at position 69 is Gln or Glu; Xaa at position 73 is Ala or Gly; Xaa at position 76 is Ser, Ala or Pro; Xaa at position 79 is Lys, Arg or Ser; Xaa at position 82 is Leu, Glu, Val or Trp; Xaa at position 85 is Leu or Val; Xaa at position 87 is Leu, Ser, Trp; Xaa at position 88 is Ala or Trp; Xaa at position 91 is Ala or Pro; Xaa at position 93 is Pro or Ser; Xaa at position 95 is His or Thr; Xaa at position 98 is His, Ile, or Thr; Xaa at position 100 is Lys or Arg; Xaa at position 101 is Asp, a or Met; Xaa at position 105 is Asn or Gln; Xaa at position 109 is Arg, Glu or Leu; Xaa at position 112 is Thr or Gln; Xaa at position 116 is Lys, Val, Trp or Ser; Xaa at position 117 is Thr or Ser; Xaa at position 120 is Asn, Gln, or His; Xaa at position 123 is Ala or Glu; with the proviso that from four to twenty-six of the amino acids designated by Xaa are different from the corresponding amino acids of native human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.

Preferred polypeptides of the present invention are those of the formula

1 5 10

[SEQ ID NO:130]

(Met

m

-Ala

n

)

p

-Asn Cys Ser Xaa Xaa Xaa Asp Glu Xaa Ile

15 20

Xaa His Leu Lys Xaa Pro Pro Xaa Pro Xaa Leu Asp Xaa

25 30 35

Xaa Asn Leu Asn Xaa Glu Asp Xaa Xaa Ile Leu Xaa Xaa

40 45

Xaa Asn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa Xaa

50 55 60

Ala Xaa Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa

65 70 75

Ile Leu Xaa Asn Xaa Xaa Pro Cys Xaa Pro Xaa Xaa Thr

80 85

Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Ile Xaa Xaa Gly

90 95 100

Asp Trp Xaa Glu Phe Arg Xaa Lys Leu Xaa Phe Tyr Leu

105 110

Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln

wherein m is 0 or 1; n is 0 or 1; p is 0 or 1; Xaa at position 4 is Asn or Ile; Xaa at position 5 is Met, Ala or Ile: Xaa at position 6 is Ile, Pro or Leu; Xaa at position 9 is Ile, Ala or Leu; Xaa at position 11 is Thr or His; Xaa at position 15 is Gln, Arg, Val or Leu; Xaa at position 18 is Leu, Ala, Asn or Arg; Xaa at position 20 is Leu or Ser; Xaa at position 23 is Phe, Pro, or Ser; Xaa at position 24 is Asn or Ala; Xaa at position 28 is Gly, Ala, Ser, Asp or Asn; Xaa at position 31 is Gln, Val, or Met; Xaa at position 32 is Asp or Ser; Xaa at position 35 is Met, Ile, Leu or Asp; Xaa at position 36 is Glu or Asp; Xaa at position 37 is Asn, Arg or Ser; Xaa at position 41 is Arg, Leu, or Thr; Xaa at position 42 is Pro or Ser; Xaa at position 45 is Glu or Leu; Xaa at position 46 is Ala or Ser; Xaa at position 48 is Asn, Val or Pro; Xaa at position 49 is Arg or His; Xaa at position 51 is Val or Ser; Xaa at position 53 is Ser, Asn, His or Gly; Xaa at position 55 is Gln or Glu; Xaa at position 59 is Ala or Gly; Xaa at position 62 is Ser, Ala or Pro; Xaa at position 65 is Lys, Arg or Ser; Xaa at position 67 is Leu, Glu, or Val; Xaa at position 68 is Leu, Glu, Val or Trp; Xaa at position 71 is Leu or Val; Xaa at position 73 is Leu, Ser or Trp; Xaa at position 74 is Ala or Trp; Xaa at position 77 is Ala or Pro; Xaa at position 79 is Pro or Ser; Xaa at position 81 is His or Thr; Xaa at position 84 is His, Ile, or Thr; Xaa at position 86 is Lys or Arg; Xaa at position 87 is Asp, Ala or Met; Xaa at position 91 is Asn or Glu; Xaa at position 95 is Arg, Glu, Leu; Xaa at position 98 Thr or Gln; Xaa at position 102 is Lys, Val, Trp or Ser; Xaa at position 103 is Thr or Ser; Xaa at position 106 is Asn, Gln, or His; Xaa at position 109 is Ala or Glu; with the proviso that from four to twenty-six of the amino acids designated by Xaa are different from the corresponding amino acids of native (15-125)human interleukin-3; or a polypeptide having substantially the same structure and substantially the same biological activity.

“Mutant amino acid sequence,” “mutant protein” or “mutant polypeptide” refers to a polypeptide having an amino acid sequence which varies from a native sequence or is encoded by a nucleotide sequence intentionally made variant from a native sequence. “Mutant protein,” “variant protein” or “mutein” means a protein comprising a mutant amino acid sequence and includes polypeptides which differ from the amino acid sequence of native hIL-3 due to amino acid deletions, substitutions, or both. “Native sequence” refers to an amino acid or nucleic acid sequence which is identical to a wild-type or native form of a gene or protein.

Human IL-3 can be characterized by its ability to stimulate colony formation by human hematopoietic progenitor cells. The colonies formed include erythroid, granulocyte, megakaryocyte, granulocytic macrophages and mixtures thereof. Human IL-3 has demonstrated an ability to restore bone marrow function and peripheral blood cell populations to therapeutically beneficial levels in studies performed initially in primates and subsequently in humans (Gillio, A. P., et al. (1990); Ganser, A, et al. (1990); Falk, S., et al. (1991). Additional activities of hIL-3 include the ability to stimulate leukocyte migration and chemotaxis; the ability to prime human leukocytes to produce high levels of inflammatory mediators like leukotrienes and histamine; the ability to induce cell surface expression of molecules needed for leukocyte adhesion; and the ability to trigger dermal inflammatory responses and fever. Many or all of these biological activities of hIL-3 involve signal transduction and high affinity receptor binding. Mutant polypeptides of the present invention may exhibit useful properties such as having similar or greater biological activity when compared to native hIL-3 or by having improved half-life or decreased adverse side effects, or a combination of these properties. They may also be useful as antagonists. hIL-3 mutant polypeptides which have little or no activity when compared to native hIL-3 may still be useful as antagonists, as antigens for the production of antibodies for use in immunology or immunotherapy, as genetic probes or as intermediates used to construct other useful hIL-3 muteins. Since hIL-3 functions by binding to its receptor(s) and triggering second messages resulting in competent signal transduction, hIL-3 muteins of this invention may be useful in helping to determine which specific amino acid sequences are responsible for these activities.

The novel hIL-3 mutant polypeptides of the present invention will preferably have at least one biological property of human IL-3 or of an IL-3-like growth factor and may have more than one IL-3-like biological property, or an improved property, or a reduction in an undesirable biological property of human IL-3. Some mutant polypeptides of the present invention may also exhibit an improved side effect profile. For example, they may exhibit a decrease in leukotriene release or histamine release when compared to native hIL-3 or (15-125) hIL-3. Such hIL-3 or hIL-3-like biological properties may include one or more of the following biological characteristics and in vivo and in vitro activities.

One such property is the support of the growth and differentiation of progenitor cells committed to erythroid, lymphoid, and myeloid lineages. For example, in a standard human bone marrow assay, an IL-3-like biological property is the stimulation of granulocytic type colonies, megakaryocytic type colonies, monocyte/macrophage type colonies, and erythroid bursts. Other IL-3-like properties are the interaction with early multipotential stem cells, the sustaining of the growth of pluripotent precursor cells, the ability to stimulate chronic myelogenous leukemia (CML) cell proliferation, the stimulation of proliferation of mast cells, the ability to support the growth of various factor-dependent cell lines, and the ability to trigger immature bone marrow cell progenitors. Other biological properties of IL-3 have been disclosed in the art. Human IL-3 also has some biological activities which may in some cases be undesirable, for example the ability to stimulate leukotriene release and the ability to stimulate increased histamine synthesis in spleen and bone marrow cultures and in vivo.

Biological activity of hIL-3 and hIL-3 mutant proteins of the present invention is determined by DNA synthesis by human acute myelogenous leukemia cells (AML). The factor-dependent cell line AML 193 was adapted for use in testing biological activity.

One object of the present invention is to provide hIL-3 muteins and hIL-3 deletion muteins with four or more amino acid substitutions in the polypeptide sequence which have similar or improved biological activity in relation to native hIL-3 or native (15-125)hIL-3.

The present invention includes mutant polypeptides comprising minimally amino acids residues 15 to 118 of hIL-3 with or without additional amino acid extensions to the N-terminus and/or C-terminus which further contain four or more amino acid substitutions in the amino acid sequence of the polypeptide. It has been found that the (15-125)hIL-3 mutant is more soluble than is hIL-3 when expressed in the cytoplasm of

E. coli,

and the protein is secreted to the periplasm in

E. coli

at higher levels compared to native hIL-3.

When expressed in the

E. coli

cytoplasm, the above-mentioned mutant hIL-3 polypeptides of the present invention may also be constructed with Met-Ala- at the N-terminus so that upon expression the Met is cleaved off leaving Ala at the N-terminus. These mutant hIL-3 polypeptides may also be expressed in

E. coli

by fusing a signal peptide to the N-terminus. This signal peptide is cleaved from the polypeptide as part of the secretion process. Secretion in

E. coli

can be used to obtain the correct amino acid at the N-terminus (e.g., Asn

15

in the (15-125) hIL-3 polypeptide) due to the precise nature of the signal peptidase. This is in contrast to the heterogeneity often observed at the N-terminus of proteins expressed in the cytoplasm in

E. coli.

The hIL-3 mutant polypeptides of the present invention may have hIL-3 or hIL-3-like activity. For example, they may possess one or more of the biological activities of native hIL-3 and may be useful in stimulating the production of hematopoietic cells by human or primate progenitor cells. The hIL-3 muteins of the present invention and pharmaceutical compositions containing them may be useful in the treatment of conditions in which hematopoietic cell populations have been reduced or destroyed due to disease or to treatments such as radiation or chemotherapy.

hIL-3 muteins of the present invention may also be useful as antagonists which block the hIL-3 receptor by binding specifically to it and preventing binding of the agonist.

One potential advantage of the (15-125) hIL-3 muteins of the present invention, particularly those which retain activity similar to or better than that of native hIL-3, is that it may be possible to use a smaller amount of the biologically active mutein to produce the desired therapeutic effect. This may make it possible to reduce the number of treatments necessary to produce the desired therapeutic effect. The use of smaller amounts may also reduce the possibility of any potential antigenic effects or other possible undesirable side effects. For example, if a desired therapeutic effect can be achieved with a smaller amount of polypeptide it may be possible to reduce or eliminate side effects associated with the administration of native IL-3 such as the stimulation of leukotriene and/or histamine release. The hIL-3 muteins of the present invention may also be useful in the activation of stem cells or progenitors which have low receptor numbers. Pharmaceutical compositions containing (15-125) hIL-3 muteins of the present invention can be administered parenterally, intravenously, or subcutaneously.

As another aspect of the present invention, there is provided a novel method for producing the novel family of human IL-3 muteins. The method of the present invention involves culturing a suitable cell or cell line, which has been transformed with a vector containing a DNA sequence coding for expression of a novel hIL-3 mutant polypeptide. Suitable cells or cell lines may be bacterial cells. For example, the various strains of

E. coli

are well-known as host cells in the field of biotechnology. Examples of such strains include

E. coli

strains JM101 [Yanish-Perron, et al. (1985)] and MON105 [Obukowicz, et al. (1992)]. Various strains of

B. subtilis

may also be employed in this method. Many strains of yeast cells known to those skilled in the art are also available as host cells for expression of the polypeptides of the present invention.

Also suitable for use in the present invention are mammalian cells, such as Chinese hamster ovary cells (CHO). General methods for expression of foreign genes in mammalian cells are reviewed in: Kaufman, R. J. (1987). High level production of proteins in mammalian cells, in

Genetic Engineering, Principles and Methods,

Vol. 9, J. K. Setlow, editor, Plenum Press, New York. An expression vector is constructed in which a strong promoter capable of functioning in mammalian cells drives transcription of a eukaryotic secretion signal peptide coding region, which is translationally fused to the coding region for the hIL-3 variant. For example, plasmids such as pcDNA I/Neo, pRc/RSV, and pRc/CMV (obtained from Invitrogen Corp., San Diego, Calif.) can be used. The eukaryotic secretion signal peptide coding region can be from the hIL-3 gene itself or it can be from another secreted mammalian protein (Bayne, M. L. et al. (1987)

Proc. Natl. Acad. Sci. USA

84, 2638-2642). After construction of the vector containing the hIL-3 variant gene, the vector DNA is transfected into mammalian cells. Such cells can be, for example, the COS7, HeLa, BHK, CHO, or mouse L lines. The cells can be cultured, for example, in DMEM media (JRH Scientific). The hIL-3 variant secreted into the media can be recovered by standard biochemical approaches following transient expression 24-72 hours after transfection of the cells or after establishment of stable cell lines following selection for neomycin resistance. The selection of suitable mammalian host cells and methods for transformation, culture, amplification, screening and product production and purification are known in the art. See, e.g., Gething and Sambrook,

Nature,

293:620-625 (1981), or alternatively, Kaufman et al,

Mol. Cell. Biol.,

5(7):1750-1759 (1985) or Howley et al., U.S. Pat. No. 4,419,446. Another suitable mammalian cell line is the monkey COS-1 cell line. A similarly useful mammalian cell line is the CV-1 cell line.

Where desired, insect cells may be utilized as host cells in the method of the present invention. See, e.g. Miller et al,

Genetic Engineering,

8:277-298 (Plenum Press 1986) and references cited therein. In addition, general methods for expression of foreign genes in insect cells using Baculovirus vectors are described in: Summers, M. D. and Smith, G. E. (1987)—A manual of methods for Baculovirus vectors and insect cell culture procedures, Texas Agricultural Experiment Station Bulletin No. 1555. An expression vector is constructed comprising a Baculovirus transfer vector, in which a strong Baculovirus promoter (such as the polyhedron promoter) drives transcription of a eukaryotic secretion signal peptide coding region, which is translationally fused to the coding region for the hIL-3 variant polypeptide. For example, the plasmid pVL1392 (obtained from Invitrogen Corp., San Diego, Calif.) can be used. After construction of the vector carrying the hIL-3 variant gene, two micrograms of this DNA is cotransfected with one microgram of Baculovirus DNA (see Summers & Smith, 1987) into insect cells, strain SF9. Pure recombinant Baculovirus carrying the hIL-3 variant is used to infect cells cultured, for example, in Excell 401 serum-free medium (JRH Biosciences, Lenexa, Kans.). The hIL-3 variant secreted into the medium can be recovered by standard biochemical approaches.

Another aspect of the present invention provides plasmid DNA vectors for use in the method of expression of these novel hIL-3 muteins. These vectors contain the novel DNA sequences described above which code for the novel polypeptides of the invention. Appropriate vectors which can transform microorganisms capable of expressing the hIL-3 muteins include expression vectors comprising nucleotide sequences coding for the hIL-3 muteins joined to transcriptional and translational regulatory sequences which are selected according to the host cells used.

Vectors incorporating modified sequences as described above are included in the present invention and are useful in the production of the hIL-3 mutant polypeptides. The vector employed in the method also contains selected regulatory sequences in operative association with the DNA coding sequences of the invention and capable of directing the replication and expression thereof in selected host cells.

Additional details may be found in WO 94/12639, which is hereby incorporated by reference in its entirety.

All references, patents or applications cited herein are incorporated by reference in their entirety.

The present invention also includes the construction and expression of (15-125)human interleukin-3 muteins having four or more amino acid substitutions in secretion vectors that optimize accumulation of correctly folded, active polypeptide. While many heterologous proteins have been secreted in

E. coli

there is still a great deal of unpredictability and limited success (Stader and Silhavy 1990). Full-length hIL-3 is such a protein, where attempts to secrete the protein in

E. coli

resulted in low secretion levels. Secretion of the variant (15-125) hIL-3 mutant polypeptides of the present invention as a fusion with a signal peptide such as LamB results in correctly folded protein that can be removed from the periplasm of

E. coli

by osmotic shock fractionation. This property of the variant (15-125) hIL-3 muteins allows for the direct and rapid screening for bioactivity of the secreted material in the crude osmotic shock fraction, which is a significant advantage. Furthermore, it provides a means of using the (15-125)hIL-3 muteins to conduct structure activity relationship (SAR) studies of the hIL-3 molecule. A further advantage of secretion of (15-125) hIL-3 muteins fused to the LamB signal peptide is that the secreted polypeptide has the correct N-terminal amino acid (Asn) due to the precise nature of the cleavage of the signal peptide by signal peptidase, as part of the secretion process.

The (15-125)hIL-3 muteins of the present invention may include hIL-3 polypeptides having Met-, Ala- or Met-Ala- attached to the N-terminus. When the muteins are expressed in the cytoplasm of

E. coli,

polypeptides with and without Met attached to the N-terminus are obtained. The methionine can in some cases be removed by methionine aminopeptidase.

Amino terminal sequences of hIL-3 muteins made in

E. coli

were determined using the method described by Hunkapillar et al., (1983). It was found that hIL-3 proteins made in

E. coli

from genes encoding Met-(15-125)hIL-3 were isolated as Met-(15-125) hIL-3. Proteins produced from genes encoding Met-Ala-(15-125) hIL-3 were produced as Ala-(15-125) hIL-3. The N-termini of proteins made in the cytoplasm of

E. coli

are affected by posttranslational processing by methionine aminopeptidase (Ben-Bassat et al., 1987) and possibly by other peptidases.

One method of creating the preferred hIL-3 (15-125) mutant genes is cassette mutagenesis [Wells, et al. (1985)] in which a portion of the coding sequence of hIL-3 in a plasmid is replaced with synthetic oligonucleotides that encode the desired amino acid substitutions in a portion of the gene between two restriction sites. In a similar manner amino acid substitutions could be made in the full-length hIL-3 gene, or genes encoding variants of hIL-3 in which from 1 to 14 amino acids have been deleted from the N-terminus and/or from 1 to 15 amino acids have been deleted from the C-terminus. When properly assembled these oligonucleotides would encode hIL-3 variants with the desired amino acid substitutions and/or deletions from the N-terminus and/or C-terminus. These and other mutations could be created by those skilled in the art by other mutagenesis methods including; oligonucleotide-directed mutagenesis [Zoller and Smith (1982, 1983, 1984), Smith (1985), Kunkel (1985), Taylor, et al. (1985), Deng and Nickoloff (1992)] or polymerase chain reaction (PCR) techniques [Saiki, (1985)].

Pairs of complementary synthetic oligonucleotides encoding portions of the amino terminus of the hIL-3 gene can be made and annealed to each other. Such pairs would have protruding ends compatible with ligation to NcoI at one end. The NcoI site would include the codon for the initiator methionine. At the other end of oligonucleotide pairs, the protruding (or blunt) ends would be compatible with a restriction site that occurs within the coding sequence of the hIL-3 gene. The DNA sequence of the oligonucleotide would encode sequence for amino acids of hIL-3 with the exception of those substituted and/or deleted from the sequence.

The NcoI enzyme and the other restriction enzymes chosen should have recognition sites that occur only once in the DNA of the plasmid chosen. Plasmid DNA can be treated with the chosen restriction endonucleases then ligated to the annealed oligonucleotides. The ligated mixtures can be used to transform competent JM101 cells to resistance to an appropriate antibiotic. Single colonies can be picked and the plasmid DNA examined by restriction analysis and/or DNA sequencing to identify plasmids with mutant hIL-3 genes.

One example of a restriction enzyme which cleaves within the coding sequence of the hIL-3 gene is ClaI whose recognition site is at codons 20 and 21. The use of ClaI to cleave the sequence of hIL-3 requires that the plasmid DNA be isolated from an

E. coli

strain that fails to methylate adenines in the DNA at GATC recognition sites. This is because the recognition site for ClaI, ATCGAT, occurs within the sequence GATCGAT which occurs at codons 19, 20 and 21 in the hIL-3 gene. The A in the GATC sequence is methylated in most

E. coli

host cells. This methylation prevents ClaI from cleaving at that particular sequence. An example of a strain that does not methylate adenines is GM48.

Interpretation of Activity of Single Amino Acid Mutants in IL-3 (15-125)

As illustrated in Tables 6 and 9, there are certain positions in the IL-3 (15-125) molecule which are intolerant of substitutions, in that most or all substitutions at these positions resulted in a considerable decrease in bioactivity. There are two likely classes of such “down-mutations”: mutations that affect overall protein structure, and mutations that interfere directly with the interaction between the IL-3 molecule and its receptor. Mutations affecting the three-dimensional structure of the protein will generally lie in the interior of the protein, while mutations affecting receptor binding will generally lie on the surface of the protein. Although the three-dimensional structure of IL-3 is unknown, there are simple algorithms which can aid in the prediction of the structure. One such algorithm is the use of “helical wheels” (Kaiser, E. T. & Kezdy, F. J., Science, 223:249-255 (1984)). In this method, the presence of alpha helical protein structures can be predicted by virtue of their amphipathic nature. Helices in globular proteins commonly have an exposed hydrophilic side and a buried hydrophobic side. As a broad generalization, in globular proteins, hydrophobic residues are present in the interior of the protein, and hydrophilic residues are present on the surface. By displaying the amino acid sequence of a protein on such a “helical wheel” it is possible to derive a model for which amino acids in alpha helices are exposed and which are buried in the core of the protein. Such an analysis of the IL-3 (15-125) molecule predicts that the following helical residues are buried in the core:

M19, I20, I23, I24, L27, L58, F61, A64, L68, A71, I74, I77, L78, L81, W104, F107, L111, Y114, L115, L118.

In addition, cysteine residues at positions 16 and 84 are linked by a disulfide bond, which is important for the overall structure or “folding” of the protein. Finally, mutations which result in a major disruption of the protein structure may be expressed at low level in the secretion system used in our study, for a variety of reasons: either because the mis-folded protein is poorly recognized by the secretion machinery of the cell; because mis-folding of the protein results in aggregation, and hence the protein cannot be readily extracted from the cells; or because the mis-folded protein is more susceptible to degradation by cellular proteases. Hence, a block in secretion may indicate which positions in the IL-3 molecule which are important for maintenance of correct protein structure.

In order to retain the activity of a variant of IL-3, it is necessary to retain both the structural integrity of the protein, and retain the specific residues important for receptor contact. Hence it is possible to define specific amino acid residues in IL-3 (15-125) which must be retained in order to preserve biological activity.

Residues predicted to be important for interaction with the receptor: D21, E22, E43, D44, L48, R54, R94, D103, K110, P113.

Residues predicted to be structurally important: C16, L58, F61, A64, I74, L78, L81, C84, P86, P92, P96, F107, L111, L115, L118.

The hIL-3 muteins of the present invention may be useful in the treatment of diseases characterized by a decreased levels of either myeloid, erythroid, lymphoid, or megakaryocyte cells of the hematopoietic system or combinations thereof. In addition, they may be used to activate mature myeloid and/or lymphoid cells. Among conditions susceptible to treatment with the polypeptides of the present invention is leukopenia, a reduction in the number of circulating leukocytes (white cells) in the peripheral blood. Leukopenia may be induced by exposure to certain viruses or to radiation. It is often a side effect of various forms of cancer therapy, e.g., exposure to chemotherapeutic drugs and of infection or hemorrhage. Therapeutic treatment of leukopenia with these hIL-3 mutant polypeptides of the present invention may avoid undesirable side effects caused by treatment with presently available drugs.

The hIL-3 muteins of the present invention may be useful in the treatment of neutropenia and, for example, in the treatment of such conditions as aplastic anemia, cyclic neutropenia, idiopathic neutropenia, Chediak-Higashi syndrome, systemic lupus erythematosus (SLE), leukemia, myelodysplastic syndrome and myelofibrosis.

Many drugs may cause bone marrow suppression or hematopoietic deficiencies. Examples of such drugs are AZT, DDI, alkylating agents and anti-metabolites used in chemotherapy, antibiotics such as chloramphenicol, penicillin and sulfa drugs, phenothiazones, tranquilizers such as meprobamate, and diuretics. The hIL-3 muteins of the present invention may be useful in preventing or treating the bone marrow suppression or hematopoietic deficiencies which often occur in patients treated with these drugs.

Hematopoietic deficiencies may also occur as a result of viral, microbial or parasitic infections and as a result of treatment for renal disease or renal failure, e.g., dialysis. The hIL-3 muteins of the present invention may be useful in treating such hematopoietic deficiency.

The treatment of hematopoietic deficiency may include administration of the hIL-3 mutein of a pharmaceutical composition containing the hIL-3 mutein to a patient. The hIL-3 muteins of the present invention may also be useful for the activation and amplification of hematopoietic precursor cells by treating these cells in vitro with the muteins of the present invention prior to injecting the cells into a patient.

Various immunodeficiencies e.g., in T and/or B lymphocytes, or immune disorders, e.g., rheumatoid arthritis, may also be beneficially affected by treatment with the hIL-3 mutant polypeptides of the present invention. Immunodeficiencies may be the result of viral infections e.g. HTLVI, HTLVII, HTLVIII, severe exposure to radiation, cancer therapy or the result of other medical treatment. The hIL-3 mutant polypeptides of the present invention may also be employed, alone or in combination with other hematopoietins, in the treatment of other blood cell deficiencies, including thrombocytopenia (platelet deficiency), or anemia. Other uses for these novel polypeptides are in the treatment of patients recovering from bone marrow transplants in vivo and ex vivo, and in the development of monoclonal and polyclonal antibodies generated by standard methods for diagnostic or therapeutic use.

Other aspects of the present invention are methods and therapeutic compositions for treating the conditions referred to above. Such compositions comprise a therapeutically effective amount of one or more of the hIL-3 muteins of the present invention in a mixture with a pharmaceutically acceptable carrier. This composition can be administered either parenterally, intravenously or subcutaneously. When administered, the therapeutic composition for use in this invention is preferably in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such a parenterally acceptable protein solution, having due regard to pH, isotonicity, stability and the like, is within the skill of the art.

The dosage regimen involved in a method for treating the above-described conditions will be determined by the attending physician considering various factors which modify the action of drugs, e.g. the condition, body weight, sex and diet of the patient, the severity of any infection, time of administration and other clinical factors. Generally, a daily regimen may be in the range of 0.2-150 μg/kg of non-glycosylated IL-3 protein per kilogram of body weight. This dosage regimen is referenced to a standard level of biological activity which recognizes that native IL-3 generally possesses an EC

50

at or about 10 picoMolar to 100 picoMolar in the AML proliferation assay described herein. Therefore, dosages would be adjusted relative to the activity of a given mutein vs. the activity of native (reference) IL-3 and it would not be unreasonable to note that dosage regimens may include doses as low as 0.1 microgram and as high as 1 milligram per kilogram of body weight per day. In addition, there may exist specific circumstances where dosages of IL-3 mutein would be adjusted higher or lower than the range of 10-200 micrograms per kilogram of body weight. These include co-administration with other CSF or growth factors: co-administration with chemotherapeutic drugs and/or radiation; the use of glycosylated IL-3 mutein; and various patient-related issues mentioned earlier in this section. As indicated above, the therapeutic method and compositions may also include co-administration with other human factors. A non-exclusive list of other appropriate hematopoietins, CSFs and interleukins for simultaneous or serial co-administration with the polypeptides of the present invention includes GM-CSF, CSF-1, G-CSF, Meg-CSF, M-CSF, erythropoietin (EPO), IL-1, IL-4, IL-2, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, LIF, B-cell growth factor, B-cell differentiation factor and eosinophil differentiation factor, stem cell factor (SCF) also known as steel factor or c-kit ligand, or combinations thereof. The dosage recited above would be adjusted to compensate for such additional components in the therapeutic composition. Progress of the treated patient can be monitored by periodic assessment of the hematological profile, e.g., differential cell count and the like.

Materials and Methods for hIL-3 Mutein Expression in

E. coli

Unless noted otherwise, all specialty chemicals were obtained from Sigma Co., (St. Louis, Mo.). Restriction endonucleases, T4 poly-nucleotides kinase,

E. coli

DNA polymerase I large fragment (Klenow) and T4 DNA ligase were obtained from New England Biolabs (Beverly, Mass.).

Escherichia coli

Strains

Strain JM101: delta (pro lac), supE, thi, F′ (traD36, rpoAB, lacI-Q, lacZdeltaM15) (Messing, 1979). This strain can be obtained from the American Type Culture Collection (ATCC), 12301 Parklawn Drive, Rockville, Md. 20852, accession number 33876. MON 105 (W3110 rpoH358) is a derivative of W3110 (Bachmann, 1972) and has been assigned ATCC accession number 55204. Strain GM48: dam-3, dcm-6, gal, ara, lac, thr, leu, tonA, tsx (Marinus, 1973) was used to make plasmid DNA that is not methylated at the sequence GATC.

Genes and Plasmids

The gene used for hIL-3 production in

E. coli

was obtained from British Biotechnology Incorporated, Cambridge, England, catalogue number BBG14. This gene is carried on a pUC based plasmid designated pP0518.

The plasmids used for production of hIL-3 in

E. coli

contain genetic elements whose use has been described (Olins et al., 1988; Olins and Rangwala, 1990). The replicon used is that of pBR327 (Covarrubias, et al., 1981) which is maintained at a copy number of about 100 in the cell (Soberon et al., 1980). A gene encoding the beta-lactamase protein is present on the plasmids. This protein confers ampicillin resistance on the cell. This resistance serves as a selectable phenotype for the presence of the plasmid in the cell.

For cytoplasmic expression vectors the transcription promoter was derived from the recA gene of

E. coli

(Sancar et al., 1980). This promoter, designated precA, includes the RNA polymerase binding site and the lexa repressor binding site (the operator). This segment of DNA provides high level transcription that is regulated even when the recA promoter is on a plasmid with the pBR327 origin of replication (Olins et al., 1988) incorporated herein by reference.

In secretion expression plasmids the transcription promoter was derived from the ara B, A, and D genes of

E. coli

(Greenfield et al., 1978). This promoter is designated pAraBAD and is contained on a 323 base pair SacII, BglII restriction fragment. The LamB secretion leader (Wong et al., 1988, Clement et al., 1981) was fused to the N-terminus of the hIL-3 gene at the recognition sequence for the enzyme NcoI (5′CCATGG3′). The hIL-3 genes used were engineered to have a HindIII recognition site (5′AAGCTT3′) following the coding sequence of the gene.

These hIL-3 variants were expressed as a fusion with the LamB signal peptide shown in

FIG. 8

, operatively joined to the araBAD promoter (Greenfield, 1978) and the g10-L ribosome binding site (Olins et al. 1988). The processed form was selectively released from the periplasm by osmotic shock as a correctly folded and fully active molecule. Secretion of (15-125) hIL-3 was further optimized by using low inducer (arabinose) concentration and by growth at 30° C. These conditions resulted in lower accumulation levels of unprocessed LamB signal peptide (15-125) hIL-3 fusion, maximal accumulation levels of processed (15-125) hIL-3 and selective release of (15-125) hIL-3 by osmotic shock fractionation. The use of a tightly regulated promoter such as araBAD from which the transcription level and hence the expression level can be modulated allowed for the optimization of secretion of (15-125) hIL-3.

The ribosome binding site used is that from gene 10 of phage T7 (Olins et al., 1988). This is encoded in a 100 base pair (bp) fragment placed adjacent to precA. In the plasmids used herein, the recognition sequence for the enzyme NcoI (CCATGG) follows the glO-L. It is at this NcoI site that the hIL-3 genes are joined to the plasmid. It is expected that the nucleotide sequence at this junction will be recognized in mRNA as a functional start site for translation (Olins et al., 1988). The hIL-3 genes used were engineered to have a HindIII recognition site (AAGCTT) downstream from the coding sequence of the gene. At this HindIII site is a 514 base pair RsaI fragment containing the origin of replication of the single stranded phage f1 (Dente et al., 1983; Olins, et al., 1990) both incorporated herein by reference. A plasmid containing these elements is pMON2341. Another plasmid containing these elements is pMON5847 which has been deposited at the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Md. 20852 under the accession number ATCC 68912.

Synthesis of Oligonucleotides

Oligonucleotides were synthesized on Nucleotide Synthesizer model 380A or 380B from Applied Biosystems, Inc. (Foster City, Calif.). Oligonucleotides were purified by polyacrylamide gel electrophoresis at concentrations from 12-20% (19:1 crosslinked) in 0.5×Tris borate buffer (0.045 M Tris, 0.045 M boric acid, 1.25 mM EDTA) followed by passage through a Nensorb column obtained from New England Nuclear (Boston, Mass.) using a PREP Automated Sample Processor obtained from DuPont, Co. (Wilmington, Del.).

Quantitation of Synthetic Oligonucleotides

Synthetic oligonucleotides were resuspended in water and quantitated by reading the absorbance at 260 nm on a Beckman DU40 Spectrophotometer (Irvine, Calif.) using a one centimeter by one millimeter quartz cuvette (Maniatis, 1982). The concentration was determined using an extinction coefficient of 1×10

4

(Voet et al., 1963; Mahler and Cordes, 1966). The oligonucleotides were then diluted to a desired concentration.

Quantitation of synthetic DNA fragments can also be achieved by adding 10 to 100 picomoles of DNA to a solution containing kinase buffer (25 mM Tris pH 8.0, 10 mM MgCl

2

, 10 mM DTT and 2 mM spermidine). To the reaction mix is added ATP to 20 micromolar, ATP radiolabeled at the gamma phosphate (5000-10,0000 dpm/pmol) and 5 units of T4 polynucleotide kinase. Radiolabelled material is obtained from New England Nuclear (Boston, Mass.). The 10 microliter mixture is incubated at 37° C. for one hour. A 1 microliter aliquot of the mixture was chromatographed on DEAE paper (Whatman) in 0.3 M ammonium bicarbonate. The counts that remained at the origin were used to determine the concentration of the synthetic DNA.

Recombinant DNA Methods

Isolation of plasmid DNA from

E. coli

cultures was performed as described (Birnboim and Doly, 1979). Some DNAs were purified by Magic™ columns, available from Promega (Madison, Wis.).

Purified plasmid DNA was treated with restriction endonucleases according to manufacturer's instructions. Analysis of the DNA fragments produced by treatment with restriction enzymes was done by agarose or polyacrylamide gel electrophoresis. Agarose (DNA grade from Fisher, Pittsburgh Pa.) was used at a concentration of 1.0% in a Tris-acetate running buffer (0.04 M Tris-acetate, 0.001M EDTA). Polyacrylamide (BioRad, Richmond Calif.) was used at a concentration of 6% (19:1 crosslinked) in 0.5×Tris-borate buffer (0.045 M Tris, 0.045 M boric acid, 1.25 mM EDTA), hereafter referred to as PAGE.

DNA polymerase I, large fragment, Klenow enzyme was used according to manufacturers instructions to catalyze the addition of mononucleotides from 5′ to 3′ of DNA fragments which had been treated with restriction enzymes that leave protruding ends. The reactions were incubated at 65° C. for 10 minutes to heat inactivate the Klenow enzyme.

The synthetic oligonucleotides were made without 5′ or 3′ terminal phosphates. In cases where such oligonucleotides were ligated end to end, the oligonucleotides were treated at a concentration of 10 picomoles per microliter with T4 polynucleotide kinase in the following buffer: 25 mM Tris, pH 8.0, 10 mM MgCl

2

, 10 mM dithiothreitol, 2 mM spermidine, 1 mM rATP. After incubation for 30 minutes at 37° C., the samples were incubated at 65° C. for five minutes to heat inactivate the kinase.

Synthetic Gene Assembly

The (15-125) hIL-3 gene was divided into four regions separated by five convenient restriction sites. In each of the four regions synthetic oligonucleotides were designed so that they would anneal in complementary pairs, with protruding single stranded ends, and when the pairs were properly assembled would result in a DNA sequence that encoded a portion of the hIL-3 gene. Amino acid substitutions in the hIL-3 gene were made by designing the oligonucleotides to encode the desired substitutions. The complementary oligonucleotides were annealed at concentration of 1 picomole per microliter in ligation buffer plus 50 mM NaCl. The samples were heated in a 100 ml beaker of boiling water and permitted to cool slowly to room temperature. One picomole of each of the annealed pairs of oligonucleotides were ligated with approximately 0.2 picomoles of plasmid DNA, digested with the appropriate restriction enzymes, in ligation buffer (25 mM Tris pH 8.0, 10 mM MgCl

2

, 10 mM dithiothreitol, 1 mM ATP, 2 mM spermidine) with T4 DNA ligase obtained from New England Biolabs (Beverly, Mass.) in a total volume of 20 μl at room temperature overnight.

DNA fragments were isolated from agarose gels by intercepting the restriction fragments on DEAE membranes from Schleicher and Schuell (Keene, N.H.) and eluting the DNA in 10 mM Tris, 1 mM EDTA, 1 M NaCl at 55° C. for 1 hour, according to manufacturer's directions. The solutions containing the DNA fragment were concentrated and desalted by using Centricon 30 concentrators from Amicon (W. R. Grace, Beverly Mass.) according to the manufacturer's directions. Ligations were performed at 15° C. overnight, except as noted, in ligation buffer.

Polymerase Chain Reaction

Polymerase Chain Reaction (hereafter referred to as PCR) techniques (Saiki, 1985) used the reagent kit and thermal cycler from Perkin-Elmer Cetus (Norwalk, Conn.). PCR is based on a thermostable DNA polymerase from

Thermus aquaticus.

The PCR technique is a DNA amplification method that mimics the natural DNA replication process in that the number of DNA molecules doubles after each cycle, in a way similar to in vivo replication. The DNA polymerase mediated extension is in a 5′ to 3′ direction. The term “primer” as used herein refers to an oligonucleotide sequence that provides an end to which the DNA polymerase can add nucleotides that are complementary to a nucleotide sequence. The latter nucleotide sequence is referred to as the “template”, to which the primers are annealed. The amplified PCR product is defined as the region comprised between the 5′ ends of the extension primers. Since the primers have defined sequences, the product will have discrete ends, corresponding to the primer sequences. The primer extension reaction was carried out using 20 picomoles (pmoles) of each of the oligonucleotides and 1 picogram of template plasmid DNA for 35 cycles (1 cycle is defined as 94 degrees C. for one minute, 50 degrees C. for two minutes and 72 degrees for three minutes.). The reaction mixture was extracted with an equal volume of phenol/chloroform (50% phenol and 50% chloroform, volume to volume) to remove proteins. The aqueous phase, containing the amplified DNA, and solvent phase were separated by centrifugation for 5 minutes in a microcentrifuge (Model 5414 Eppendorf Inc, Fremont Calif.). To precipitate the amplified DNA the aqueous phase was removed and transferred to a fresh tube to which was added 1/10 volume of 3M NaOAc (pH 5.2) and 2.5 volumes of ethanol (100% stored at minus 20 degrees C.). The solution was mixed and placed on dry ice for 20 minutes. The DNA was pelleted by centrifugation for 10 minutes in a microcentrifuge and the solution was removed from the pellet. The DNA pellet was washed with 70% ethanol, ethanol removed and dried in a speedvac concentrator (Savant, Farmingdale, N.Y.). The pellet was resuspended in 25 microliters of TE (20 mM Tris-HCl pH 7.9, 1 mM EDTA). Alternatively the DNA was precipitated by adding equal volume of 4M NH

4

OAc and one volume of isopropanol [Treco et al., (1988)]. The solution was mixed and incubated at room temperature for 10 minutes and centrifuged. These conditions selectively precipitate DNA fragments larger than ˜20 bases and were used to remove oligonucleotide primers. One quarter of the reaction was digested with restriction enzymes [Higuchi, (1989)] an on completion heated to 70 degrees C. to inactivate the enzymes.

Recovery of Recombinant Plasmids from Ligation Mixes

E. coli

JM101 cells were made competent to take up DNA. Typically, 20 to 100 ml of cells were grown in LB medium to a density of approximately 150 Klett units and then collected by centrifugation. The cells were resuspended in one half culture volume of 50 mM CaCl

2

and held at 4° C. for one hour. The cells were again collected by centrifugation and resuspended in one tenth culture volume of 50 mM CaCl

2

. DNA was added to a 150 microliter volume of these cells, and the samples were held at 4° C. for 30 minutes. The samples were shifted to 42° C. for one minute, one milliliter of LB was added, and the samples were shaken at 37° C. for one hour. Cells from these samples were spread on plates containing ampicillin to select for transformants. The plates were incubated overnight at 37° C. Single colonies were picked, grown in LB supplemented with ampicillin overnight at 37° C. with shaking. From these cultures DNA was isolated for restriction analysis.

Culture Medium

LB medium (Maniatis et al., 1982) was used for growth of cells for DNA isolation. M9 minimal medium supplemented with 1.0% casamino acids, acid hydrolyzed casein, Difco (Detroit, Mich.) was used for cultures in which recombinant hIL-3 was produced. The ingredients in the M9 medium were as follows: 3 g/liter KH

2

PO

4

, 6 g/l Na

2

HPO

4

, 0.5 g/l NaCl, 1 g/l NH

4

Cl, 1.2 mM MgSO

4

, 0.025 mM CaCl

2

, 0.2% glucose (0.2% glycerol with the AraBAD promoter), 1% casamino acids, 0.1 ml/l trace minerals (per liter 108 g FeCl

3

.6H

2

O, 4.0 g ZnSO

4

.7H

2

O, 7.0 CoCl

2

.2H

2

O, 7.0 g Na

2

MoO

4

.2H

2

O, 8.0 g CuSO

4

.5H

2

O, 2.0 g H

3

BO

3

, 5.0 g MnSO

4

.H

2

O, 100 ml concentrated HCl). Bacto agar was used for solid media and ampicillin was added to both liquid and solid LB media at 200 micrograms per milliliter.

DNA Sequence Analysis

The nucleotide sequencing of plasmid DNA was determined using a Genesis 2000 sequencer obtained from DuPont (Wilmington, Del.) according to the methods of Prober et al. (1987) and Sanger et al. (1977). Some DNA sequences were performed using Sequenase™ polymerase (U.S. Biochemicals, Cleveland, Ohio) according to manufacturer's directions.

Production of Recombinant hIL-3 Muteins in

E. coli

with Vectors Employing the recA Promoter

E. coli

strains harboring the plasmids of interest were grown at 37° C. in M9 plus casamino acids medium with shaking in a Gyrotory water bath Model G76 from New Brunswick Scientific (Edison, N.J.). Growth was monitored with a Klett Summerson meter (green 54 filter), Klett Mfg. Co. (New York, N.Y.). At a Klett value of approximately 150, an aliquot of the culture (usually one milliliter) was removed for protein analysis. To the remaining culture, nalidixic acid (10 mg/ml) in 0.1 N NaOH was added to a final concentration of 50 μg/ml. The cultures were shaken at 37° C. for three to four hours after addition of nalidixic acid. A high degree of aeration was maintained throughout the bacterial growth in order to achieve maximal production of the desired gene product. The cells were examined under a light microscope for the presence of refractile bodies (RBs). One milliliter aliquots of the culture were removed for analysis of protein content.

Production of Recombinant hIL-3 Proteins from the pAraBAD Promoter in

E. coli

E. coli

strains harboring the plasmids of interest were grown at 30° C. with shaking in M9 medium plus casamino acids and glycerol. Growth was monitored with a Klett Summerson calorimeter, using a green 54 filter. At a Klett value of about 150, an aliquot of the culture (usually one milliliter) was removed for protein analysis. To the remaining culture, 20% arabinose was added to a final concentration of 0.05%. The cultures were shaken at 30° C. for three to four hours after addition of arabinose. A high degree of aeration was maintained throughout the bacterial growth in order to achieve maximal production of the desired gene product. One milliliter aliquots of the culture were removed for analysis of protein content.

Secretion and Osmotic Shock

Three hour post induction samples were fractionated by osmotic shock [Neu and Heppel (1965)]. The optical density (Klett value) of the cultures was determined and 1 ml of cells were centrifuged in a Sigma microcentrifuge (West Germany) model 202MK in 1.5 mls snap top microcentrifuge tubes for 5 minutes at 10,000 rpm. The cell pellet was resuspended very gently by pipeting in a room temperature sucrose solution (20% sucrose w/v, 30 mM Tris-Hcl pH7.5, 1 mM EDTA), using 1 μl Klett unit. Following a 10 minute incubation at room temperature, the cells were centrifuged for 5 minutes at 10,000 rpm. The sucrose fraction was carefully removed from the cell pellet. The cell pellet was then resuspended very gently by pipeting in ice cold distilled water, using 1 μl/1 Klett unit. Following a 10 minute incubation on ice, the cells were centrifuged for 5 minutes at 12,000 rpm. The water fraction was carefully removed. Equal volumes of the sucrose and water fractions were pooled and aliquoted to provide samples for activity screening.

Analysis of Protein Content of

E. coli

Cultures Producing hIL-3 Mutant Polypeptides

Bacterial cells from cultures treated as described above were collected from the medium by centrifugation. Aliquots of these cells were resuspended in SDS loading buffer (4×: 6 g SDS, 10 ml beta-mercaptoethanol, 25 ml upper Tris gel stock (0.5 M Tris HCl pH 6.8, 0.4% SDS) brought to 50 ml with glycerol, 0.2% bromophenol blue was added) at a concentration of one microliter per Klett unit. These samples were incubated at 85° C. for five minutes and vortexed. Five or ten microliter aliquots of these samples were loaded on 15% polyacrylamide gels prepared according to the method of Laemmli (1970). Protein bands were visualized by staining the gels with a solution of acetic acid, methanol and water at 5:1:5 ratio (volume to volume) to which Coomassie blue had been added to a final concentration of 1%. After staining, the gels were washed in the same solution without the Coomassie blue and then washed with a solution of 7% acetic acid, 5% methanol. Gels were dried on a gel drier Model SE1160 obtained from Hoeffer (San Francisco, Calif.). The amount of stained protein was measured using a densitometer obtained from Joyce-Loebl (Gateshead, England). The values obtained were a measure of the amount of the stained hIL-3 protein compared to the total of the stained protein of the bacterial cells.

Western Blot Analysis of hTL-3 Muteins made in

E. coli

In some

E. coli

cultures producing hIL-3, the level of accumulation of the hIL-3 protein is lower than 5% of total bacterial protein. To detect hIL-3 produced at this level, Western blot analysis was used. Proteins from cultures induced with nalidixic acid or arabinose were run on polyacrylamide gels as described above except that volumes of sample loaded were adjusted to produce appropriate signals. After electrophoresis, the proteins were electroblotted to APT paper, Transa-bind, Schleicher and Schuell (Keene, N.H.) according to the method of Renart et al. (1979). Antisera used to probe these blots had been raised in rabbits, using peptides of the sequence of amino acids 20 to 41 and 94 to 118 of hIL-3 as the immunogens. The presence of bound antibody was detected with Staphylococcal protein A radiolabeled with

125

I, obtained from New England Nuclear (Boston, Mass.).

Fractionation of

E. coli

Cells Producing hIL-3 Proteins in the Cytoplasm

Cells from

E. coli

cultures harboring plasmids that produce hIL-3 muteins were induced with nalidixic acid. After three hours, the hIL-3 muteins accumulated in refractile bodies. The first step in purification of the hIL-3 muteins was to sonicate cells. Aliquots of the culture were resuspended from cell pellets in sonication buffer: 10 mM Tris, pH 8.0, 1 mM EDTA, 50 mM NaCl and 0.1 mM PMSF. These resuspended cells were subjected to several repeated sonication bursts using the microtip from a Sonicator cell disrupter, Model W-375 obtained from Heat Systems-Ultrasonics Inc. (Farmingdale, N.Y.). The extent of sonication was monitored by examining the homogenates under a light microscope. When nearly all of the cells had been broken, the homogenates were fractionated by centrifugation. The pellets, which contain most of the refractile bodies, are highly enriched for hIL-3 muteins.

Methods: Extraction, Refolding and Purification of Interleukin-3 (IL-3) Muteins Expressed as Refractile Bodies in

E. coli

Extraction of Refractile Bodies (RB's)

For each gram of RB's (and typically one gram is obtained from a 300 ml

E. coli

culture), 5 ml of a solution containing 6M guanidine hydrochloride (GnHCl), 50 mM 2-N-cyclohexylaminoethanesulfonic acid (CHES) ph 9.5 and 20 mM dithiothreitol (DTT) was added. The RB's were extracted with a Bio-Homogenizer for 15-30 seconds and gently rocked for 2 hours at 5 degrees centigrade (5° C.) to allow the protein to completely reduce and denature.

Refolding of the IL-3 Muteins

The protein solution was transferred to dialysis tubing (1000 molecular weight cut-off) and dialyzed against at least 100 volumes of 4M GnHCl—50 mM CHES pH 8.0. The dialysis was continued overnight at 5° C. while gently stirring. Subsequently dialysis was continued against at least 100 volumes of 2M GnHCl—50 mM CHES pH 8.0 and dialyzed overnight at 5° C. while gently stirring.

Purification of the IL-3 Muteins

The protein solution was removed from the dialysis tubing and acidified by the addition of 40% acetonitrile (CH

3

CN)—0.2% trifluoroacetic acid (TFA) to a final concentration of 20% CH

3

CN—0.1% TFA. This was centrifuged (16,000×g for 5 minutes) to clarify and the supernatant was loaded onto a Vydac C-18 reversed phase column (10×250 mm) available from Vydac (Hesperia, Calif.) previously equilibrated in 20% CH

3

CN—0.1% TFA. The column was eluted with a linear gradient (0.2% CH

3

CN/minute) between 40-50% CH

3

CN—0.1% TFA at a flow rate of 3 ml/minute while collecting 1.5 ml fractions. The fractions were analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and the appropriate fractions pooled. The pooled material was dried by lyophilization or in a Speed Vac concentrator. The dry powder was reconstituted with 10 mM ammonium bicarbonate pH 7.5, centrifuged (16,000×g for 5 minutes) to clarify and assayed for protein concentration by the method of Bradford (1976) with bovine serum albumin as the standard. Such protein can be further analyzed by additional techniques such as, SDS-PAGE, electrospray mass spectrometry, reverse phase HPLC, capillary zone electrophoresis, amino acid composition analysis, and ELISA (enzyme-linked immunosorbent assay).

hIL-3 Sandwich ELISA

IL-3 protein concentrations can be determined using a sandwich ELISA based on an affinity purified polyclonal goat anti-rhIL-3. Microtiter plates (Dynatech Immulon II) were coated with 150 μl goat-anti-rhIL-3 at a concentration of approximately 1 μg/ml in 100 mM NaHCO3, pH 8.2. Plates were incubated overnight at room temperature in a chamber maintaining 100% humidity. Wells were emptied and the remaining reactive sites on the plate were blocked with 200 μl of solution containing 10 mM PBS, 3% BSA and 0.05% Tween 20, pH 7.4 for 1 hour at 37° C. and 100% humidity. Wells were emptied and washed 4× with 150 mM NaCl containing 0.05% Tween 20 (wash buffer). Each well then received 150 μl of dilution buffer (10 mM PBS containing 0.1% BSA, 0.01% Tween 20, pH 7.4), containing rhIL-3 standard, control, sample or dilution buffer alone. A standard curve was prepared with concentrations ranging from 0.125 ng/ml to 5 ng/ml using a stock solution of rhIL-3 (concentration determined by amino acid composition analysis). Plates were incubated 2.5 hours at 37° C. and 100% humidity. Wells were emptied and each plate was washed 4× with wash buffer. Each well then received 150 μl of an optimal dilution (as determined in a checkerboard assay format) of goat anti-rhIL-3 conjugated to horseradish peroxidase. Plates were incubated 1.5 hours at 37° C. and 100% humidity. Wells were emptied and each plate was washed 4× with wash buffer. Each well then received 150 ul of ABTS substrate solution (Kirkegaard and Perry). Plates were incubated at room temperature until the color of the standard wells containing 5 ng/ml rhIL-3 had developed enough to yield an absorbance between 0.5-1.0 when read at a test wavelength of 410 nm and a reference wavelength of 570 nm on a Dynatech microtiter plate reader. Concentrations of immunoreactive rhIL-3 in unknown samples were calculated from the standard curve using software supplied with the plate reader.

AML Proliferation Assay for Bioactive Human Interleukin-3

The factor-dependent cell line AML 193 was obtained from the American Type Culture Collection (ATCC, Rockville, Md.). This cell line, established from a patient with acute myelogenous leukemia, is a growth factor dependent cell line which displayed enhanced growth in GM/CSF supplemented medium (Lange, B., et al., (1987); Valtieri, M., et al., (1987). The ability of AML 193 cells to proliferate in the presence of human IL-3 has also been documented. (Santoli, D., et al., (1987)). A cell line variant was used, AML 193 1.3, which was adapted for long term growth in IL-3 by washing out the growth factors and starving the cytokine dependent AML 193 cells for growth factors for 24 hours. The cells were then replated at 1×10

5

cells/well in a 24 well plate in media containing 100 U/ml IL-3. It took approximately 2 months for the cells to grow rapidly in IL-3. These cells were maintained as AML 193 1.3 thereafter by supplementing tissue culture medium (see below) with human IL-3.

AML 193 1.3 cells were washed 6 times in cold Hanks balanced salt solution (HBSS, Gibco, Grand Island, N.Y.) by centrifuging cell suspensions at 250×g for 10 minutes followed by decantation of supernatant. Pelleted cells were resuspended in HBSS and the procedure was repeated until six wash cycles were completed. Cells washed six times by this procedure were resuspended in tissue culture medium at a density ranging from 2×10

5

to 5×10

5

viable cells/ml. This medium was prepared by supplementing Iscove's modified Dulbecco's Medium (IMDM, Hazleton, Lenexa, Kans.) with albumin, transferrin, lipids and 2-mercaptoethanol. Bovine albumin (Boehringer-Mannheim, Indianapolis, Ind.) was added at 500 μg/ml; human transferrin (Boehringer-Mannheim, Indianapolis, Ind.) was added at 100 μg/ml; soybean lipid (Boehringer-Mannheim, Indianapolis, Ind.) was added at 50 μg/ml; and 2-mercaptoethanol (Sigma, St. Louis, Mo.) was added at 5×10

−5

M.

Serial dilutions of human interleukin-3 or human interleukin-3 variant protein (hIL-3 mutein) were made in triplicate series in tissue culture medium supplemented as stated above in 96 well Costar 3596 tissue culture plates. Each well contained 50 μl of medium containing interleukin-3 or interleukin-3 variant protein once serial dilutions were completed. Control wells contained tissue culture medium alone (negative control). AML 193 1.3 cell suspensions prepared as above were added to each well by pipetting 50 μl (2.5×10

4

cells) into each well. Tissue culture plates were incubated at 37° C. with 5% CO

2

in humidified air for 3 days. On day 3, 0.5 μCi

3

H-thymidine (2 Ci/mM, New England Nuclear, Boston, Mass.) was added in 50 μl of tissue culture medium. Cultures were incubated at 37° C. with 5% CO

2

in humidified air for 18-24 hours. Cellular DNA was harvested onto glass filter mats (Pharmacia LKB, Gaithersburg, Md.) using a TOMTEC cell harvester (TOMTEC, Orange, Conn.) which utilized a water wash cycle followed by a 70% ethanol wash cycle. Filter mats were allowed to air dry and then placed into sample bags to which scintillation fluid (Scintiverse II, Fisher Scientific, St. Louis, Mo. or BetaPlate Scintillation Fluid, Pharmacia LKB, Gaithersburg, Md.) was added. Beta emissions of samples from individual tissue culture wells were counted in a LKB Betaplate model 1205 scintillation counter (Pharmacia LKB, Gaithersburg, Md.) and data was expressed as counts per minute of

3

H-thymidine incorporated into cells from each tissue culture well. Activity of each human interleukin-3 preparation or human interleukin-3 variant preparation was quantitated by measuring cell proliferation (

3

H-thymidine incorporation) induced by graded concentrations of interleukin-3 or interleukin-3 variant. Typically, concentration ranges from 0.05 pM-10

5

pM are quantitated in these assays. Activity is determined by measuring the dose of interleukin-3 or interleukin-3 variant which provides 50% of maximal proliferation [EC

50

=0.5×(maximum average counts per minute of

3

H-thymidine incorporated per well among triplicate cultures of all concentrations of interleukin-3 tested—background proliferation measured by

3

H-thymidine incorporation observed in triplicate cultures lacking interleukin-3]. This EC

50

value is also equivalent to 1 unit of bioactivity. Every assay was performed with native interleukin-3 as a reference standard so that relative activity levels could be assigned.

Relative biological activities of IL-3 muteins of the present invention are shown in Table 1. The Relative Biological Activity of IL-3 mutants is calculated by dividing the EC

50

of (1-133) hIL-3 by the EC

50

of the mutant. The Relative Biological Activity may be the average of replicate assays.

TABLE 1

BIOLOGICAL ACTIVITY OF IL-3 MUTEINS

Relative*

Plasmid

Polypeptide

Biological

Code

Structure

Activity

Reference (1-133)hIL-3

1

pMON13298

SEQ ID NO. 82

3

pMON13299

SEQ ID NO. 83

2

pMON13300

SEQ ID NO. 84

3

pMON13301

SEQ ID NO. 85

2

pMoN13302

SEQ ID NO. 86

1.2

pMON13303

SEQ ID NO. 87

0.6

pMON13287

SEQ ID NO. 88

26

pMON13288

SEQ ID NO. 89

24

pMON13289

SEQ ID NO. 90

13

pMON13290

SEQ ID NO. 91

20

pMON13292

SEQ ID NO. 92

6

pMON13294

SEQ ID NO. 93

3

pMON13295

SEQ ID NO. 94

3

pMON13312

SEQ ID NO. 95

4

pMON13313

SEQ ID NO. 96

8

pMON13285

SEQ ID NO. 259

32

pMON13286

SEQ ID NO. 260

8

pMON13325

SEQ ID NO. 261

8

pMON13326

SEQ ID NO. 262

25

pMON13330

SEQ ID NO. 263

19

pMON13329

SEQ ID NO. 406

10

pMON13364

SEQ ID NO. 117

13

pMON13475

SEQ ID NO. 280

7

pMON13366

SEQ ID NO. 281

38

pMON13367

SEQ ID NO. 282

36

pMON13368

SEQ ID NO. 278

1.6

pMON13369

SEQ ID NO. 283

10

pMON13370

SEQ ID NO. 284

6

pMON13373

SEQ ID NO. 285

12

pMON13374

SEQ ID NO. 286

6

pMON13375

SEQ ID NO. 287

14

pMON13376

SEQ ID NO. 288

0.4

pMON13377

SEQ ID NO. 289

0.4

pMON13379

SEQ ID NO. 291

0.9

pMON13380

SEQ ID NO. 279

0.05

pMON13381

SEQ ID NO. 293

10

pMON13382

SEQ ID NO. 313

38

pMON13383

SEQ ID NO. 294

0.5

pMON13384

SEQ ID NO. 295

0.25

pMON13385

SEQ ID NO. 292

1

pMON13387

SEQ ID NO. 308

32

pMON13388

SEQ ID NO. 296

23

pMON13389

SEQ ID NO. 297

10

pMON13391

SEQ ID NO. 298

30

pMON13392

SEQ ID NO. 299

17

pMON13393

SEQ ID NO. 300

32

pMON13394

SEQ ID NO. 301

20

pMON13395

SEQ ID NO. 302

11

pMON13396

SEQ ID NO. 303

20

pMON13397

SEQ ID NO. 304

16

pMON13398

SEQ ID NO. 305

36

pMON13399

SEQ ID NO. 306

18

pMON13404

SEQ ID NO. 307

1.3

pMON13417

SEQ ID NO. 310

24

pMON13420

SEQ ID NO. 311

19

pMON13421

SEQ ID NO. 331

0.5

pMON13432

SEQ ID NO. 312

10

pMON13400

SEQ ID NO. 317

0.09

pM0N13402

SEQ ID NO. 318

20

pMON13403

SEQ ID NO. 321

0.03

pMON13405

SEQ ID NO. 267

9

pMON13406

SEQ ID NO. 264

5

pMON13407

SEQ ID NO. 266

16

pMON13408

SEQ ID NO. 269

7

pMON13409

SEQ ID NO. 270

15

pMON13410

SEQ ID NO. 271

0.4

pMON13411

SEQ ID NO. 322

1.2

pMON13412

SEQ ID NO. 323

0.5

pMON13413

SEQ ID NO. 324

0.6

pMON13414

SEQ ID NO. 265

4

pMON13415

SEQ ID NO. 268

4

pMON13418

SEQ ID NO. 326

0.5

pMON13419

SEQ ID NO. 325

0.015

pMON13422

SEQ ID NO. 272

0.4

pMON13423

SEQ ID NO. 273

0.4

pMON13424

SEQ ID NO. 274

3

pMON13425

SEQ ID NO. 275

6

pMON13426

SEQ ID NO. 276

>0.0003

pMON13429

SEQ ID NO. 277

>0.0002

pMoN13440

SEQ ID NO. 319

9

pMON13451

SEQ ID NO. 320

0.1

pMON13459

SEQ ID NO. 328

0.003

pMON13416

SEQ ID NO. 309

19.9

pMON13428

SEQ ID NO. 327

0.008

pMON13467

SEQ ID NO. 329

0.16

pMON13446

SEQ ID NO. 315

21.5

pMON13390

SEQ ID NO. 316

20

*The Relative Biological Activity of IL-3 mutants is calculated by dividing the EC

50

of (1-133) hIL-3 by the EC

50

of the mutant.

The following assay is used to measure IL-3 mediated sulfidoleukotriene release from human mononuclear cells.

IL-3 Mediated Sulfidoleukotriene Release from Human Mononuclear Cells

Heparin-containing human blood was collected and layered onto an equal volume of Ficoll-Paque (Pharmacia # 17-0840-02) ready to use medium (density 1.077 g/ml.). The Ficoll was warmed to room temperature prior to use and clear 50 ml polystyrene tubes were utilized. The Ficoll gradient was spun at 300×g for 30 minutes at room temperature using a H1000B rotor in a Sorvall RT6000B refrigerated centrifuge. The band containing the mononuclear cells was carefully removed, the volume adjusted to 50 mls with Dulbecco's phosphate-buffered saline (Gibco Laboratories cat. # 310-4040PK), spun at 400×g for 10 minutes at 4° C. and the supernatant was carefully removed. The cell pellet was washed twice with HA Buffer [20 mM Hepes (Sigma # H-3375), 125 mM NaCl (Fisher # S271-500), 5 mM KCl (Sigma # P-9541), 0.5 mM glucose (Sigma # G-5000), 0.025% Human Serum Albumin (Calbiochem # 126654) and spun at 300×g, 10 min., 4° C. The cells were resuspended in HACM Buffer (HA buffer supplemented with 1 mM CaCl2 (Fisher # C79-500) and 1 mM MgCl2 (Fisher # M-33) at a concentration of 1×106 cells/ml and 180 μl were transferred into each well of 96 well tissue culture plates. The cells were allowed to acclimate at 37° C. for 15 minutes. The cells were primed by adding 10 μls of a 20× stock of various concentrations of cytokine to each well (typically 100000, 20000, 4000, 800, 160, 32, 6.4, 1.28, 0 fM IL3). The cells were incubated for 15 minutes at 37° C. Sulfidoleukotriene release was activated by the addition of 10 μls of 20× (1000 nM) fmet-leu-phe (Calbiochem # 344252) final concentration 50 nM FMLP and incubated for 10 minutes at 37° C. The plates were spun at 350×g at 4° C. for 20 minutes. The supernatants were removed and assayed for sulfidoleukotrienes using Cayman's Leukotriene C4 EIA kit (Cat. #420211) according to manufacturers' directions. Native (15-125)hIL-3 was run as a standard control in each assay.

Native hIL-3 possesses considerable inflammatory activity and has been shown to stimulate synthesis of the arachidonic acid metabolites LTC

4

, LTD

4

, and LTE

4

; histamine synthesis and histamine release. Human clinical trials with native hIL-3 have documented inflammatory responses (Biesma, et al., BLOOD, 80:1141-1148 (1992) and Postmus, et al., J. CLIN. ONCOL., 10:1131-1140 (1992)). A recent study indicates that leukotrienes are involved in IL-3 actions in vivo and may contribute significantly to the biological effects of IL-3 treatment (Denzlinger, C., et al., BLOOD, 81:2466-2470 (1993))

Some muteins of the present invention may have an improved therapeutic profile as compared to native hIL-3 or (15-125)hIL-3. For example, some muteins of the present invention may have a similar or more potent growth factor activity relative to native hIL-3 or (15-125)hIL-3 without having a similar or corresponding increase in the stimulation of leukotriene or histamine. These muteins would be expected to have a more favorable therapeutic profile since the amount of polypeptide which needs to be given to achieve the desired growth factor activity (e. g. cell proliferation) would have a lesser leukotriene or histamine stimulating effect. In studies with native hIL-3, the stimulation of inflammatory factors has been an undesirable side effect of the treatment. Reduction or elimination of the stimulation of mediators of inflammation would provide an advantage over the use of native hIL-3.

The pMON13288 polypeptide has demonstrated a more potent growth factor activity relative to native hIL-3 in the AML 193 cell proliferation assay (EC

50

=0.8-3.8 pM for pMON13288 and EC

50

=30.2 pM for native hIL-3) without demonstrating a corresponding increase in the stimulation of leukotriene C

4

(LTC

4

) production and histamine release, i. e., it stimulated LTC

4

production and histamine release with a potency similar to that of native hIL-3 while having an improved ability to stimulate cell proliferation compared to native hIL-3. Thus with the pMON13288 polypeptide it would be expected that one would be able to produce a desired therapeutic response, e. g., cell proliferation, with less stimulation of the undesirable inflammatory mediators.

Some muteins of the present invention have antigenic profiles which differ from that of native hIL-3. For example, in a competition ELISA with an affinity purified polyclonal goat anti-hIL-3 antibody, native hIL-3 significantly blocked the binding of labeled hIL-3 to polyclonal anti-hIL-3 antibody whereas the pMON13288 polypeptide failed to block the binding of hIL-3 to anti-hIL-3 antibody.

Table 2 lists the sequences of some oligonucleotides used in making the muteins of the present invention.

Table 3 lists the amino acid sequence of native (15-125)hIL-3 (Peptide #1) and the amino acid sequences of some mutant polypeptides of the present invention. The sequences are shown with the amino acid numbering corresponding to that of native hIL-3 [FIG.

1

].

Table 4 lists the nucleotide sequences of some DNA sequences which encode mutant polypeptides of the present invention.

TABLE 2

OLIGONUCLEOTIDES

Oligo #1

Length: 000040

CATGGCTAAC TGCTCTATAA TGATCGATGA AATTATACAT

[SEQ ID NO:15]

Oligo #2

Length: 000045

CTTTAAGTGA TGTATAATTT CATCGATCAT TATAGAGCAG TTAGC

[SEQ ID NO:16]

Oligo #3

Length: 000036

CACTTAAAGA GACCACCTGC ACCTTTGCTG GACCCG

[SEQ ID NO:17]

Oligo #4

Length: 000036

GAGGTTGTTC GGGTCCAGCA AAGGTGCAGG TGGTCT

[SEQ ID NO:18]

Oligo #5

Length: 000036

CACTTAAAGA GACCACCTAA CCCTTTGCTG GACCCG

[SEQ ID NO:19]

Oligo #6

Length: 000036

GAGGTTGTTC GGGTCCAGCA AAGGGTTAGG TGGTCT

[SEQ ID NO:20]

Oligo #7

Length: 000036

CACTTAAAGG TTCCACCTGC ACCTTTGCTG GACAGT

[SEQ ID NO:21]

Oligo #8

Length: 000036

GAGGTTGTTA CTGTCCAGCA AAGGTGCAGG TGGAAC

[SEQ ID NO:22]

Oligo #9

Length: 000027

AACAACCTCA ATGCTGAAGA CGTTGAT

[SEQ ID NO:23]

Oligo #10

Length: 000018

ATCAACGTCT TCAGCATT

[SEQ ID NO:24]

Oligo #11

Length: 000027

AACAACCTCA ATTCTGAAGA CATGGAT

[SEQ ID NO:25]

Oligo #12

Length: 000018

ATCCATGTCT TCAGAATT

[SEQ ID NO:26]

Oligo #13

Length: 000022

CATGGGAACC ATATGTCAGG AT

[SEQ ID NO:27]

Oligo #14

Length: 000018

ATCCTGACAT ATGGTTCC

[SEQ ID NO:28]

Oligo #15

Length: 000016

TGAACCATAT GTCAGG

[SEQ ID NO:29]

Oligo #16

Length: 000024

AATTCCTGAC ATATGGTTCA TGCA

[SEQ ID NO:30]

Oligo #17

Length: 000020

AATTCGAACC ATATGTCAGA

[SEQ ID NO:31]

Oligo #18

Length: 000020

AGCTTCTGAC ATATGGTTCG

[SEQ ID NO:32]

Oligo #19

Length: 000022

ATCGAACCAT ATGTCAGATG CA

[SEQ ID NO:33]

Oligo #20

Length: 000018

TCTGACATAT GGTTCGAT

[SEQ ID NO:34]

Oligo #21

Length: 000036

ATCCTGATGG AACGAAACCT TCGACTTCCA AACCTG

[SEQ ID NO:35]

Oligo #22

Length: 000027

AAGTCGAAGG TTTCGTTCCA TCAGGAT

[SEQ ID NO:36]

Oligo #23

Length: 000036

ATCCTGATGG AACGAAACCT TCGAACTCCA AACCTG

[SEQ ID NO:37]

Oligo #24

Length: 000027

AGTTCGAAGG TTTCGTTCCA TCAGGAT

[SEQ ID NO:38]

Oligo #25

Length: 000024

CTCGCATTCG TAAGGGCTGT CAAG

[SEQ ID NO:39]

Oligo #26

Length: 000024

CCTTACGAAT GCGAGCAGGT TTGG

[SEQ ID NO:40]

Oligo #27

Length: 000024

GAGAGCTTCG TAAGGGCTGT CAAG

[SEQ ID NO:41]

Oligo #28

Length: 000024

CCTTACGAAG CTCTCCAGGT TTGG

[SEQ ID NO:42]

Oligo #29

Length: 000015

CACTTAGAAA ATGCA

[SEQ ID NO:43]

Oligo #30

Length: 000020

TTTTCTAAGT GCTTGACAGC

[SEQ ID NO:44]

Oligo #31

Length: 000015

AACTTAGAAA ATGCA

[SEQ ID NO:45]

Oligo #32

Length: 000020

TTTTCTAAGT TCTTGACAGC

[SEQ ID NO:46]

Oligo #33

Length: 000048

GGTGATTGGA TGTCGAGAGG GTGCGGCCGT GGCAGAGGGC AGACATGG

[SEQ ID NO:47]

Oligo #34

Length: 000048

CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCA CCATCAAG

[SEQ ID NO:48]

Oligo #35

Length: 000048

GATGATTGGA TGTCGAGAGG GTGCGGCCGT GGCAGAGGGC AGACATGG

[SEQ ID NO:49]

Oligo #36

Length: 000048

CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCA TCATCAAG

[SEQ ID NO:50]

Oligo #37

Length: 000018

TACGAGATTA CGAAGAAT

[SEQ ID NO:51]

Oligo #38

Length: 000018

CGTAATCTCG TACCATGT

[SEQ ID NO:52]

Oligo #39

Length: 000018

TTGGAGATTA CGAAGAAT

[SEQ ID NO:53]

Oligo #40

Length: 000018

CGTAATCTCC AACCATGT

[SEQ ID NO:54]

Oligo #41

Length: 000019

TGCCTCAATA CCTGATGCA

[SEQ ID NO:55]

Oligo #42

Length: 000021

TCAGGTATTG AGGCAATTCT T

[SEQ ID NO:56]

Oligo #43

Length: 000026

AATTCTTGCC AGTCACCTGC CTTGAT

[SEQ ID NO:57]

Oligo #44

Length: 000016

GCAGGTGACT GCCAAG

[SEQ ID NO:58]

Oligo #45

Length: 000032

AATTCCGGGA AAAACTGACG TTCTATCTGG TT

[SEQ ID NO:59]

Oligo #46

Length: 000037

CTCAAGGGAA ACCAGATAGA ACGTCAGTTT TTCCCGG

[SEQ ID NO:60]

Oligo #47

Length: 000032

ACCCTTGAGC ACGCGCAGGA ACAACAGTAA TA

[SEQ ID NO:61]

Oligo #48

Length: 000027

AGCTTATTAC TGTTGTTCCT GCGCGTG

[SEQ ID NO:62]

Oligo #49

Length: 000032

ACCCTTGAGC AAGCGCAGGA ACAACAGTAA TA

[SEQ ID NO:63]

Oligo #50

Length: 000027

AGCTTATTAC TGTTGTTCCT GCGCTTG

[SEQ ID NO:64]

Oligo #51

Length: 000034

GCCGATACCGCGGCATACTCCCACCATTCAGAGA

[SEQ ID NO:155]

Oligo #52

Length: 000033

GCCGATAAGATCTAAAACGGGTATGGAGAAACA

[SEQ ID NO:156]

Oligo #53

ATAGTCTTCCCCAGATATCTAACGCTTGAG

[SEQ ID NO:157]

Oligo #54

Length: 24

CAATACCTGATGCGTTTTCTAAGT

[SEQ ID NO:158]

Oligo #55

Length: 33

GGTTTCGTTCCATCAGAATGTCCATGTCTTCAG

[SEQ ID NO:159]

Oligo #165

NCOECRV1.REQ Length: 000040

CATGGCTAAC TGCTCTAACA TGATCGATGA AATTATAACA

[SEQ ID NO:162]

Oligo #166

NCOECRV4.REQ Length: 000045

CTTTAAGTGT GTTATAATTT CATCGATCAT GTTAGAGCAG TTAGC

[SEQ ID NO:163]

Oligo #167

NCOECRV2.REQ Length: 000036

CACTTAAAGC AGCCACCTTT GCCTTTGCTG GACTTC

[SEQ ID NO:164]

Oligo #168

NCOECRV5.REQ Length: 000036

GAGGTTGTTG AAGTCCAGCA AAGGCAAAGG TGGCTG

[SEQ ID NO:165]

Oligo #169

2D5M6SUP.REQ Length: 000027

AACAACCTCA ATGACGAAGA CATGTCT

[SEQ ID NO:166]

Oligo #170

2D5M6SLO.REQ Length: 000018

AGACATGTCT TCGTCATT

[SEQ ID NO:167)

Oligo #15(A)

Length: 000016

TGAACCATAT GTCAGG

[SEQ ID NO:168]

Oligo #16(A)

Length: 000024

AATTCCTGAC ATATGGTTCA TGCA

[SEQ ID NO:169]

Oligo #B1

19ALA1.REQ Length: 000040

CATGGCAAAC TGCTCTATAG CTATCGATGA AATTATACAT

[SEQ ID NO:170]

Oligo #B2

19ALA4.REQ Length: 000045

CTTTAAGTGA TGTATAATTT CATCGATAGC TATAGAGCAG TTTGC

[SEQ ID NO:171]

Oligo #B3

191LE1.REQ Length: 000040

CATGGCAAAC TGCTCTATAA TCATCGATGA AATTATACAT

[SEQ ID NO:172]

Oligo #B4

191LE4.REQ Length: 000045

CTTTAAGTGA TGTATAATTT CATCGATGAT TATAGAGCAG TTTGC

[SEQ ID NO:173]

Oligo #B5

49ASP1.REQ Length: 000036

ATCCTGGACG AACGAARCCT TCGAACTCCA AACCTG

[SEQ ID NO:174]

Oligo #B6

49ASP4.REQ Length: 000027

AGTTCGAAGG TTTCGTTCGT CCAGGAT

[SEQ ID NO:175]

Oligo #B7

49ILE1.REQ Length: 000036

ATCCTGATCG AACGAAACCT TCGARCTCCA AACCTG

[SEQ ID NO:176]

Oligo #B8

49ILE4.REQ Length: 000027

AGTTCGAAGG TTTCGTTCGA TCAGGAT

[SEQ ID NO:177]

Oligo #B9

49LEU1.REQ Length: 000036

ATCCTGCTGG AACGAARCCT TCGAACTCCA AACCTG

[SEQ ID NO:178]

Oligo #B1O

49LEU4.REQ Length: 000027

AGTTCGAAGG TTTCGTTCCA GCAGGAT

[SEQ ID NO:179]

Oligo #B11

42S45V3.REQ Length: 000027

AACAACCTCA ATTCTGAAGA CGTTGAT

[SEQ ID NO:180]

Oligo #B12

42S45V6.REQ Length: 000018

ATCARCGTCT TCAGARTT

[SEQ ID NO:181]

Oligo #B13

18I23A5H.REQ Length: 000051

CGCGCCATGG CTAACTGCTC TATAATGATC GATGAAGCAA TACATCACTTA

[SEQ ID NO:182]

Oligo #B14

2341HIN3.REQ Length: 000018

CGCGTCGATA AGCTTATT

[SEQ ID NO:183]

Oligo #B15

2341NC0.REQ Length: 000018

GGAGATATAT CCATGGCT

[SEQ ID NO:184]

Oligo #B16

2A5M6S0D.REQ Length: 000042

TCGGTCCATC AGAATAGACA TGTCTTCAGC ATTGAGGTTG TT

[SEQ ID NO:185]

Oligo #B17

2A5V6S0D.REQ Length: 000042

TCGGTCCATC AGAATAGAAA CGTCTTCAGC ATTGAGGTTG TT

[SEQ ID NO:186]

Oligo #B18

2D5M6S0D.REQ Length: 000042

TCGGTCCATC AGAATAGACA TGTCTTCGTC ATTGAGGTTG TT

[SEQ ID NO:187]

Oligo #B19

2D5V6S0D.REQ Length: 000042

TCGGTCCATC AGAATAGAAA CGTCTTCGTC ATTGAGGTTG TT

[SEQ ID NO:188]

Oligo #B20

2S5M650D.REQ Length: 000042

TCGGTCCATC AGAATAGACA TGTCTTCAGA ATTGAGGTTG TT

[SEQ ID NO:189]

Oligo #B21

2S5V6S0D.REQ Length: 000042

TCGGTCCATC AGAATAGAAA CGTCTTCAGA ATTGAGGTTG TT

[SEQ ID NO:190]

Oligo #B22

100ARG3.REQ Length: 000048

CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCA TCATCCGT

[SEQ ID NO:191]

Oligo #B23

100ARG8.REQ Length: 000026

AATTCTTGCC AGTCACCTGC ACGGAT

[SEQ ID NO:192]

Oligo #B24

101MET4.REQ Length: 000016

ATGGGTGACT GGCAAG

[SEQ ID NO:193]

Oligo #B25

10R01M8.REQ Length: 000026

AATTCTTGCC AGTCACCCAT ACGGAT

[SEQ ID NO:194]

Oligo #B26

23ALA1.REQ Length: 000040

CATGGCTAAC TGCTCTATTA TGATCGATGA AGCAATACAT

[SEQ ID NO:195]

Oligo #B27

23ALA4.REQ Length: 000045

CTTTAAGTGA TGTATTGCTT CATCGATCAT AATAGAGCAG TTAGC

[SEQ ID NO:196]

Oligo #B28

29V2R4S2.REQ Length: 000036

CACTTAAAGG TACCACCTCG CCCTTCCCTG GACCCG

[SEQ ID NO:197]

Oligo #B29

29V2R4S5.REQ Length: 000036

GAGGTTGTTC GGGTCCAGGG AAGGGCGAGG TGGTAC

[SEQ ID NO:198]

Oligo #B30

34SER2.REQ Length: 000036

CACTTAAAGA GACCACCTGC ACCTTCCCTG GACCCG

[SEQ ID NO:199]

Oligo #B31

34SER5.REQ Length: 00003E

GAGGTTGTTC GGGTCCAGGG AAGGTGCAGG TGGTCT

[SEQ ID NO:200]

Oligo #B32

42D45M3.REQ Length: 000027

AACAACCTCA ATGACGAAGA CATGGAT

[SEQ ID NO:201]

Oligo #B33

42D45M6.REQ Length: 000018

ATCCATGTCT TCGTCATT

[SEQ ID NO:202]

Oligo #B34

42D45V3.REQ Length: 000027

AACAACCTCA ATGACGAAGA CGTCGAT

[SEQ ID NO:203]

Oligo #B35

42D45V6.REQ Length: 000018

ATCGACGTCT TCGTCATT

[SEQ ID NO:204]

Oligo #B36

42D5M6S3.REQ Length: 000027

AACAACCTCA ATGACGAAGA CATGTCT

[SEQ ID NO:205]

Oligo #B37

42D5M6S6.REQ Length: 000018

AGACATGTCT TCGTCATT

[SEQ ID NO:206]

Oligo #B38

42D5V6S3.REQ Length: 000027

AACAACCTCA ATGACGAAGA CGTCTCT

[SEQ ID NO:207]

Oligo #B39

42D5V6S6.REQ Length: 000018

AGAGACGTCT TCGTCATT

[SEQ ID NO:208]

Oligo #B40

50ASP1.REQ Length: 000036

ATCCTGATGG ACCGAAACCT TCGACTTCCA AACCTG

[SEQ ID NO:209]

Oligo #B41

50ASP4.REQ Length: 000027

AAGTCGAAGG TTTCGGTCCA TCAGGAT

[SEQ ID NO:210]

Oligo #B42

50D56S1.REQ Length: 000036

ATCCTGATGG ACCGAAACCT TCGACTTAGC AACCTG

[SEQ ID NO:211]

Oligo #B43

56SER5.REQ Length: 000024

CCTTACGAAG CTCTCCAGGT TGCT

[SEQ ID NO:212]

Oligo #B44

82TRP2.REQ Length: 000018

CGTAATCTCT GGCCATGT

[SEQ ID NO:213]

Oligo #B45

82TRP6.REQ Length: 000018

CCAGAGATTA CGAAGAAT

[SEQ ID NO:214]

Oligo #B46

9E12Q6W1.REQ Length: 000032

AATTCCGGGA AAAACTGCAA TTCTATCTGT GG

[SEQ ID NO:215]

Oligo #B47

9E12Q6W3.REQ Length: 000037

CTCAAGGGTC CACAGATAGA ATTGCAGTTT TTCCCGG

[SEQ ID NO:216]

Oligo #B48

9E12Q6V1.REQ Length: 000032

AATTCCGGGA AAAACTGCAA TTCTATCTGG TT

[SEQ ID NO:217]

Oligo #B49

9E12Q6V3.REQ Length: 000037

CTCAAGGGTA ACCAGATAGA ATTGCAGTTT TTCCCGG

[SEQ ID NO:218]

Oligo #B50

S09E16V1.REQ Length: 000023

AATTCCGGGA AAAACTGACG TTC

[SEQ ID NO:219]

Oligo #B51

S09E16V3.REQ Length: 000028

AACCAGATAG AACGTCAGTT TTTCCCGG

[SEQ ID NO:220]

Oligo #B52

S116VD31.REQ Length: 000023

TATCTGGTTA CCCTTGAGTA ATA

[SEQ ID NO:221]

Oligo #B53

SECR1D33.REQ Length: 000018

AGCTTATTAC TTCAAGGGT

[SEQ ID NO:222]

Oligo #B54

S9E2Q6V1.REQ Length: 000023

AATTCCGGGA AAAACTGCAA TTC

[SEQ ID NO:223]

Oligo #B55

S9E2Q6V3.REQ Length: QQOQ28

AACCAGATAG AATTGCAGTT TTTCCCGG

[SEQ ID NO:224]

Oligo #B56

Ent338.Lo Length: 61

[SEQ ID NO:225]

CGATCATTAT AGAGCAGTTA GCCTTGTCAT CGTCGTCCTT GTAATCAGTT

TCTGGATATG C

Oligo #B57

Ent338.UP Length: 63

CATGGCATAT CCAGAAACTG ATTACAAGGA CGACGATGAC AAGGCTAACT

SEQ ID NO:226]

GCTCTATAAT GAT

09L2Q6S1.REQ Length: 000032

AATTCCGGCT TAAACTGCCA TTCTATCTGT CT

[SEQ ID NO:227]

09L2Q6S3.REQ Length: 000037

CTCAAGGGTA GACAGATAGA ATTGCAGTTT AAGCCGG

[SEQ ID NO:228]

117S2.REQ Length: 000032

TCTCTTGAGC AAGCGCAGGA ACAACAGTAA TA

[SEQ ID NO:229]

19I0L3A1.REQ Length: 000040

CATGGCAAAC TGCTCTATAA TACTCGATGA AGCAATACAT

[SEQ ID NO:230]

19I0L3A4.REQ Length: 000045

CTTTAAGTGA TGTATTGCTT CATCGAGTAT TATAGAGCAG TTTGC

[SEQ.ID NO.:231]

20P23A1.REQ Length: 000040

CATGGCAAAC TGCTCTATAA TGCCAGATGA AGCAATACAT

[SEQ. ID NO.:232]

20P23A4.REQ Length: 000045

CTTTAAGTGA TGTATTGCTT CATCTGGCAT TATAGAGCAG TTTGC

[SEQ. ID NO.:233]

23L1.REQ Length: 000040

CATGGCaAAC TGCTCTATAA TGATCGATGA AactgATACAT

[SEQ. ID NO.:234]

23L4.REQ Length: 000045

CTTTAAGTGA TGTATcagTT CATCGATCAT TATAGAGCAG TTtGC

[SEQ. ID NO.:235]

29I4S7S2.REQ Length: 000036

CACTTAAAGA TACCACCTAA CCCTAGCCTG GACAGT

[SEQ. ID NO.:236]

29I4S7S5.REQ Length: 000036

GAGGTTAGCA CTGTCCAGGC TAGGGTTAGG TGGTAT

[SEQ. ID NO.:237]

38A5V6S3.REQ Length: 000027

GCTAACCTCA ATTCCGAAGA CGTCTCT

[SEQ. ID NO.:238]

38A5V6S6.REQ Length: 000018

AGAGACGTCT TCGGAATT

[SEQ. ID NO.:239]

50D51S1.REQ Length: 000036

ATCCTGATGG ACTCCAACCT TCGAACTCCA AACCTG

[SEQ. ID NO.:240]

50D5154.REQ Length: 000027

AGTTCGAAGG TTGGAGTCCA TCAGGAT

[SEQ. ID NO.:241]

5VYWPTT3.REQ Length: 000048

GTTCCCTATT GGACGGCCCC TCCCTCTCGA ACACCAATCA CGATCAAG

[SEQ. ID NO.:242]

5VYWPTT7.REQ Length: 000049

CGTGATTGGT GTTCGAGAGG GAGGGGCCGT CCAATAGGGA ACACATGG

[SEQ. ID NO.:243]

62P3H5S2.REQ Length: 000024

CTCGCATTCC CACATGCTTC TAAG

[SEQ. ID NO.:244]

62P63H2.REQ Length: 000024

CTCGCATTCC CACATGCTGT CAAG

[SEQ. ID NO.:245]

62P63H5.REQ Length: 000024

ATGTGGGAAT GCGAGCAGGT TTGG

[SEQ. ID NO.:246]

65S67Q6.REQ Length: 000020

TTTTCTAATT GCTTAGAAGC

[SEQ. ID NO.:247]

67Q3.REQ Length: 000015

CAATTAGAAA ATGCA

[SEQ. ID NO.:248]

67Q6.REQ Length: 000023

TTTTCTAATT GCTTGACAGC

[SEQ. ID NO.:249]

76P1.REQ Length: 000021

TCAGGTATTG AGCCAATTCT T

[SEQ. ID NO.:250]

76P5.REQ Length: 000019

TGGCTCAATA CCTGATGCA

[SEQ. ID NO.:251]

79S2.REQ Length: 000018

TCTAATCTCC AACCATGT

[SEQ. ID NO.:252)

79S6.REQ Length: 000018

TTGGAGATTA GAAAGAAT

[SEQ. ID NO.:253]

9L2Q67S3.REQ Length: 000037

CTCAAGAGAA GACAGATAGA ATTGCAGTTT AAGCCGG

[SEQ. ID NO.:254]

9LQS1181.REQ Length: 000043

AATTCCGGCT TAAACTGCAA TTCTATCTGT CTACCCTTTA ATA

[SEQ. ID NO.:256]

9LQS1183.REQ Length: 000043

AGCTTATTAA AGGGTAGACA GATAGAATTG CAGTTTAAGC CGG

[SEQ. ID NO.:257]

S9L2Q6S1.REQ Length: 000043

AATTCCGGCT TAAACTGCAA TTCTATCTGT CTACCCTTTA ATA

[SEO. ID NO.:258]

TABLE 3

POLYPEPTIDES

The numbering of the mutated amino acid positions which describe the following polypeptides corresponds to the sequence of the full-length, native (1-133)hIL-3.

PEPTIDE #1:

pMON5988 (Example 43), (15-125)hIL-3

SEQ ID NO: 65;

PEPTIDE #2:

pMON13344 (Example 8), (15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A and 45V)

SEQ ID NO: 66;

PEPTIDE #3:

pMON13345 (Example 9), (15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S and 45M)

SEQ ID NO: 67;

PEPTIDE #4:

pMON13346 (Example 10), (15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S and 45M)

SEQ ID NO: 68;

PEPTIDE #5:

pMON13347 (Example 12), (15-125)hIL-3 (51R, 55L, 59L, 62V, 67N and 69E)

SEQ ID NO: 69;

PEPTIDE #6:

pMON13348 (Example 13), (15-125)hIL-3 (51R, 55L, 60S, 62V, 67N and 69E)

SEQ ID NO: 70;

PEPTIDE #7:

pMON13349 (Example 14), (15-125)hIL-3 (51R, 55T, 59L, 62V, 67H and 69E)

SEQ ID NO: 71;

PEPTIDE #8:

pMON13350 (Example 16), (15-125)hIL-3 (73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A and 105Q)

SEQ ID NO: 72;

PEPTIDE #9:

pMON13355 (Example 17), (15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A and 105Q)

SEQ ID NO: 73;

PEPTIDE #10:

pMON13352 (Example 19), (15-125)hIL-3 (109E, 116V, 120Q and 123E)

SEQ ID NO: 74;

PEPTIDE #11:

pMON13354 (Example 20), (15-125)hIL-3 (109E, 116V, 117S, 120H and 123E)

SEQ ID NO: 75;

PEPTIDE #12:

pMON13360 (Example 21), (15-125)hIL-3 (73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

SEQ. ID. NO: 76;

PEPTIDE #13:

pMON13361 (Example 22), (15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E),

SEQ ID NO: 77;

PEPTIDE #14:

pMON13362 (Example 23), (15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E)

SEQ ID NO: 78;

PEPTIDE #15:

pMON13363 (Example 24), (15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N and 69E)

SEQ ID NO: 79;

PEPTIDE #16:

pMON13364 (Example 25), (15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H and 69E)

SEQ ID NO: 80;

PEPTIDE #17:

pMON13365 (Example 26), (15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N and 69E)

SEQ ID NO: 81;

PEPTIDE #18:

pMON13298 (Example 27), Met-Ala-(15-125)hIL-3 (73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

SEQ ID NO: 82;

PEPTIDE #19:

pMON13299 (Example 28), Met-Ala-(15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E);

SEQ ID NO: 83;

PEPTIDE #20:

pMON13300 (Example 29), Met-Ala-(15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E)

SEQ ID NO: 84;

PEPTIDE #21:

pMON13301 (Example 30), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N and 69E)

SEQ ID NO: 85;

PEPTIDE #22:

pMON13302 (Example 31), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H and 69E)

SEQ ID NO: 86;

PEPTIDE #23:

pMON13303 (Example 32), Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N and 69E)

SEQ ID NO: 87;

PEPTIDE #24:

pMON13287 (Example 33), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 88;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #25:

pMON13288 (Example 34), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 89;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #26:

pMON13289 (Example 35), Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 90;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #27:

pMON13290 (Example 36), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T,

SEQ ID NO: 91;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #28:

pMON13292 (Example 37), Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T,

SEQ ID NO: 92;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #29:

pMON13294 (Example 38), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T,

SEQ ID NO: 93;

101A, 105Q, 109E, 116V, 117S, 120H and 123E)

PEPTIDE #30:

pMON13295 (Example 39), Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T,

SEQ ID NO: 94;

101A, 105Q, 109E, 116V, 117S, 120H and 123E)

PEPTIDE #31:

pMON13312 (Example 40), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T,

SEQ ID NO: 95;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #32:

pMON13313 (Example 41), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T,

SEQ ID NO: 96;

101A, 105Q, 109E, 116V, 117S, 120H and 123E)

PEPTIDE #A3:

pMON13285 (Example 60), Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 50D)

SEQ ID NO: 259;

PEPTIDE #A4:

pMON13286 (Example 61), Met-Ala-(15-125)hIL-3 (42D, 45M, 46S)

SEQ ID NO: 260;

PEPTIDE #A5:

pMON13325 (Example 62), Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 116W)

SEQ ID NO: 261;

PEPTIDE #A6:

pMON13326 (Example 63), Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 50D, 116W)

SEQ ID NO: 262;

PEPTIDE #A7:

pMON13330 (Example 65), Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 50D, 95R, 98I, 100R, 116W)

SEQ ID NO: 263;

PEPTIDE #A8:

pMON13329 (Example 66), Met-Ala-(15-125)hIL-3, (42D, 45M, 46S, 98I, 100R, 116W)

SEQ ID NO: 406;

PEPTIDE #B1:

pMON13406 (Example 69), Met-Ala-(15-125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G,

SEQ ID NO: 264;

76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B2:

pMON13414 (Example 70), Met-Ala-(15-125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 265;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B3:

pMON13407 (Example 71), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45V, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 266;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B4:

pMON13405 (Example 72), Met-Ala-(15-125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 267;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B5:

pMON13415 (Example 73), Met-Ala-(15-125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 268;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B6:

pMON13408 (Example 74), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 49I, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 269;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B7:

pMON13409 (Example 75), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 49L, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 270;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B8:

pMON13410 (Example 76), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 49D, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 271;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B9:

pMON13422 (Example 77), Met-Ala-(15-125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 49I, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 272;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B10:

pMON13423 (Example 78), Met-Ala-(15-125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 49I, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 273;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B11:

pMON13424 (Example 79), Met-Ala-(15-125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 49L, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 274;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B12:

pMON13425 (Example 80), Met-Ala-(15-125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 49L, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 275;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B13:

pMON13426 (Example 81), Met-Ala-(15-125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 49D, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 276;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B14:

pMON13429 (Example 82), Met-Ala-(15-125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 49D, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 277;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B15:

pMON13368 (Example 83), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 278;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B16:

pMON13380 (Example 84), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 279;

101A, 105Q, 109E, 112Q, 116V, 120Q and 123E)

PEPTIDE #B17:

pMON13475 (Example 86), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 280;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B18:

pMON13366 (Example 87), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42N, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 281;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B19:

pMON13367 (Example 88), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 282;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B20:

pMON13369 (Example 89), Met-Ala-(15-125)hIL-3-(18I, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 283;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B21:

pMON13370 (Example 90), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45M, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 284;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B22:

pMON13373 (Example 91), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 285;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B23:

pMON13374 (Example 92), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42S, 45M, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 286;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B24:

pMON13375 (Example 93), Met-Ala-(15-119)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,)

SEQ ID NO: 287;

PEPTIDE #B25:

pMON13376 (Example 94), Met-Asp-(15-119)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 288;

101A, 105Q, 109E, 112Q, 116V)

PEPTIDE #B26:

pMON13377 (Example 95), Met-Ala-(15-119)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 289;

101A, 105Q, 109E, 112Q, 116V)

PEPTIDE #B27:

pMON13378 (Example 96), Met-Asp-(15-119)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 290;

101A, 105Q, 109E, 116V)

PEPTIDE #B28:

pMON13379 (Example 97), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 291;

101A, 105Q, 109E, 112Q, 116V, 120Q and 123E)

PEPTIDE #B29:

pMON13385 (Example 98), Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32R, 34S, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 292;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B30:

pMON13381 (Example 99), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82W, 87S, 93S, 98I,

SEQ ID NO: 293;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B31:

pMON13383 (Example 100), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 294;

101A, 105Q, 109E, 112Q, 116V, 120Q and 123E)

PEPTIDE #B32:

pMON13384 (Example 101), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 295;

101A, 105Q, 109E, 112Q, 116V, 120Q and 123E)

PEPTIDE #B33:

pMON13388 (Example 102), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 50D, 51R, 55L, 56S, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 296;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B34:

pMON13389 (Example 103), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 297;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B35:

pMON13391 (Example 104), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 34S, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 298;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B36:

pMON13392 (Example 105), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A,, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 299;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B37:

pMON13393 (Example 106), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45M, 46S, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 300;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B38:

pMON13394 (Example 107), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 46S, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 301;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B39:

pMON13395 (Example 108), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29V, 32R, 34S, 37P, 42D, 45V, 46S, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 302;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B40:

pMON13396 (Example 109), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 303;

101A, 105Q, 109E, 100R, 101M, 116V, 120Q and 123E)

PEPTIDE #B41:

pMON13397 (Example 110), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82W, 87S, 93S, 98I,

SEQ ID NO: 304;

101A, 105Q, 109E, 100R, 101M, 116V, 120Q and 123E)

PEPTIDE #B42:

pMON13398 (Example 111), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 305;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B43:

pMON13399 (Example 112), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29V, 32R, 34S, 37P, 42D, 45V, 46S, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 306;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B44:

pMON13404 (Example 113), Met-Ala-(15-119)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 307;

101A, 105Q, 109E, 112Q, 116V)

PEPTIDE #B45:

pMON13387 (Example 114), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 308;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B46:

pMON13416 (Example 115), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 309;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B47:

pMON13417 (Example 116), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 310;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B48:

pMON13420 (Example 117), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 56S, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 311;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B49:

pMON13421 (Example 118), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45M, 46S, 50D, 51R, 55L, 56S, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 331;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B50:

pMON13432 (Example 119), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45M, 46S, 50D, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 312;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B51:

pMON13382 (Example 120), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 313;

101A, 105Q, 109E, 112Q, 116W, 120Q and 123E)

PEPTIDE #B52:

pMON13476 (Example 85), Met-Asp-(15-125)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 314;

101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B53:

pMON13446 (Example 121), Met-Ala-Tyr-Pro-Glu-Thr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V,

SEQ ID NO: 315;

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #B54:

pMON13390 (Example 122), Met-Ala-Tyr-Pro-Glu-Thr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,

SEQ ID NO: 316;

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #C-2:

pMON13400 (Example 124), Met-Ala-(15-125)hIL-3 (18I, 20P, 23A, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R, 55T, 59L, 62V, 67H, 69E, 73G,

SEQ ID NO: 317;

76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #C-3:

pMON13402 (Example 125), Met-Ala-(15-125)hIL-3 (18I, 23L, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A,

SEQ ID NO: 318;

79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #C-10:

pMON13440 (Example 131), Met-Ala-(15-125)hIL-3 (18I, 23A, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A,

SEQ ID NO: 319;

79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #C-11:

pMON13451 (Example 132), Met-Ala-(15-125)hIL-3 (18I, 19I, 20L, 23A, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R, 55T, 59L, 62V, 67H, 69E,

SEQ ID NO: 320;

73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #C-4:

pMON13403 (Example 126), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 50D, 51S, 55T, 59L, 62P, 63H, 67Q, 69E, 73G, 76A, 79R, 82Q,

SEQ ID NO: 321;

87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

PEPTIDE #C-5:

pMON13411 (Example 127), Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 322;

101A, 105Q, 109L, 112Q, 116S, 120Q and 123E)

PEPTIDE #C-6:

pMON13412 (Example 128), Met-Ala-(15-118)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

SEQ ID NO: 323.

101A, 105Q, 109L, 112Q, 116S)

TABLE 4

DNA SEQUENCES

The numbering of the mutated amino acid positions in the

modified hIL-3 proteins corresponds to the sequence of the

full-length, native (1-133)hIL-3.

DNA Sequence #1: pMON13287 (Example 33), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E,

73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and

123E) SEQ ID NO:97;

DNA Sequence #2: pMON13290 (Example 36), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E,

73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and

123E) SEQ ID NO:98;

DNA Sequence #3: pMON13313 (Example 41), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E,

73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H

and 123E) SEQ ID NO:98;

DNA Sequence #4: pMON13288 (Example 34), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E,

73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and

123E) SEQ ID NO:100;

DNA Sequence #5: pMON13312 (Example 40), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E,

73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and

123E) SEQ ID NO:101;

DNA Sequence #6: pMON13294 (Example 38), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E,

73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H

and 123E) SEQ ID NO:102;

DNA Sequence #7: pMON13289 (Example 35), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E,

73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and

123E) SEQ ID NO:103;

DNA Sequence #8: pMON13292 (Example 37), encoding Met-Ala-(15-125)hIL-

3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E,

73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and

123E) SEQ ID NO:104;

DNA Sequence #9: pMON13295 (Example 39), encodlng Met-Ala-(15-125)hIL-

3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E,

73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H

and 123E) SEQ ID NO:105;

DNA Sequence #10: pMON13344 (Example 8), (15-125)hIL-3 (18I, 25H, 29R,

32A, 37p, 42A and 45V) SEQ ID NO:106;

DNA Sequence #11: pMON13345 (Example 9), (15-125)hIL-3 (18I, 25H, 29R,

32N, 37P, 42S and 45M) SEQ ID NO:107;

DNA Sequence #12: pMON13346 (Example 10), (15-125)hIL-3 (18I, 25H,

29V, 32A, 37S, 42S and 45M) SEQ ID NO:108;

DNA Sequence #13: pMON13347 (Example 12), (15-125)hIL-3 (51R, 55L,

59L, 62V, 67N and 69E) SEQ ID NO:109;

DNA Sequence #14: pMON13348 (Example 13), (15-125)hIL-3 (51R, 55L,

60S, 62V, 67N and 69E) SEQ ID N0:110;

DNA Sequence #15: pMON13349 (Example 14), (15-125)hIL-3 (51R, 55T,

59L, 62V, 67H and 69E) SEQ ID NO:111;

DNA Sequence #16: pMON13350 (Example 16), (15-125)hIL-3 (73G, 76A,

79R, 82Q, 87S, 93S, 98I, 101A and 105Q) SEQ ID NO:112;

DNA Sequence #17: pMON13355 (Example 17), (15-125)hIL-3 (73G, 76A,

79R, 82V, 87S, 93S, 98T, 101A and 105Q) SEQ ID NO:113;

DNA Sequence #18: pMON13352 (Example 19), (15-125)hIL-3 (109E, 116V,

120Q and 123E) SEQ ID NO:114;

DNA Sequence #19: pMON13354 (Example 20), (15-125)hIL-3 (109E, 116V,

117S, 120H and 123E) SEQ ID NO:115;

DNA Sequence #20: pMON13363 (Example 24), (15-125)hIL-3 (18I, 25H,

29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N and 69E)

SEQ ID NO:116;

DNA Sequence #21: pMON13364 (Example 25), (15-125)hIL-3 (18I, 25H,

29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H and 69E)

SEQ ID NO:117;

DNA Sequence #22: pMON13365 (Example 26), (15-125)hIL-3 (18I, 25H,

29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N and 69E)

SEQ ID NO:118;

DNA Sequence #23: pM0N13360 (Example 21), (15-125)hIL-3 (73G, 76A,

79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E),

SEQ ID NO:119;

DNA Sequence #24: pMON13361 (Example 22), (15-125)hIL-3 (73G, 76A,

79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E),

SEQ ID NO:120;

DNA Sequence 125: pMON13362 (Example 23), (15-125)hIL-3 (73G, 76A,

79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E)

SEQ ID NO:121;

DNA Sequence #26: pMON13301 (Example 30), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N

and 69E) SEQ ID NO:122;

DNA Sequence #27: pMON13302 (Example 31), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H

and 69E) SEQ ID NO:123;

DNA Sequence #28: pMON13303 (Example 32), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N

and 69E) SEQ ID NO:124;

DNA Sequence #29: pMON13298 (Example 27), encoding Met-Ala-(15-

125)hIL-3 (73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:125;

DNA Sequence #30: pMON13299 (Example 28), encoding Met-Ala-(15-

125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,

120Q and 123E); SEQ ID NO:126;

DNA Sequence #31: pMON13300 (Example 29), encoding Met-Ala-(15-

125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,

117S, 120H and 123E) SEQ ID No:127;

DNA Sequence #33: pMON13438 (Example 59), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q

and 123E) SEQ ID NO:161;

DNA Sequence #A3: pMON13285 (Example 60), encoding Met-Ala-(15-

125)hIL-3 (42D, 45M, 46S, 50D) SEQ ID NO:398;

DNA Sequence #A4: pMON13286 (Example 61), encoding Met-Ala-(15-

125)hIL-3 (42D, 45M, 46S) SEQ ID NO:399;

DNA Sequence #B1: pMON13406 (Example 69), encoding Met-Ala-(15-

125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:332;

DNA Sequence #B2: pMON13414 (Example 70), encoding Met-Ala-(15-

125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:333;

DNA Sequence #B3: pMON13407 (Example 71), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45V, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q

and 123E) SEQ ID NO:334;

DNA Sequence #B4: pMON13405 (Example 72), encoding Met-Ala-(15-

125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:335;

DNA Sequence #B5: pMON13415 (Example 73), encoding Met-Ala-(15-

125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:336;

DNA Sequence #B6: pMON13408 (Example 74), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 49I, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:337;

DNA Sequence #B7: pMON13409 (Example 75), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 49L, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:338;

DNA Sequence #B8: pMON13410 (Example 76), encoding,Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 49D, 51R, 55T, 59L, 62V,

67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:339;

(

DNA Sequence #B9: pMON13422 (Example 77), encoding Met-Ala-(15-

125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 49I, 51R, 55T, 59L,

62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:340;

DNA Sequence #B10: pMON13423 (Example 78), encoding Met-Ala-(15-

125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 49I, 51R, 55T, 59L,

62V, 67H, 69E, 73G, 76A, 79R, 82Q; 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 23E) SEQ ID NO:341;

DNA Sequence #B11: pMON13424 (Example 79), encoding Met-Ala-(15-

125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 49L, 51R, 55T, 59L,

62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:342;

DNA Sequence #B12: pMON13425 (Example 80),encoding Met-Ala-(15-

125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 49L, 51R, 55T, 59L,

62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:343;

DNA Sequence #B13: pMON13426 (Example 81), encoding Met-Ala-(15-

125)hIL-3 (18I, 19A, 25H, 29R, 32N, 37P, 42S, 45V, 49D, 51R, 55T,

59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:344;

DNA Sequence #B14: pMON13429 (Example 82), encoding Met-Ala-(15-

125)hIL-3 (18I, 19I, 25H, 29R, 32N, 37P, 42S, 45V, 49D, 51R, 55T,

59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:345;

DNA Sequence #B15: pMON13368 (Example 83), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:346;

DNA Sequence #B16: pMON13380 (Example 84), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 112Q,

116V, 120Q and 123E) SEQ ID NO:347;

DNA Sequence #B17: pMON13475 (Example 86), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L,

60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:348;

DNA Sequence #B18: pMON13366 (Example 87), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42N, 45V, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:349;

DNA Sequence #B19: pMON13367 (Example 88), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:350;

DNA Sequence #B20: pMON13369 (Example 89), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:351;

DNA Sequence #B21: pMON13370 (Example 90), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45M, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:352;

DNA Sequence #B22: pMON13373 (Example 91), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:353;

DNA Sequence #B23: pMON13374 (Example 92), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42S, 45M, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:354;

DNA Sequence #B24: pMON13375 (Example 93), encoding Met-Ala-(15-

119)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,) SEQ

ID NO:355;

DNA Sequence #B25: pMON13376 (Example 94), Met-Asp-(15-119)hIL-3 (18I,

23A, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G,

76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 112Q, 116V) SEQ ID

NO:356;

DNA Sequence #B26: pMON13377 (Example 95), encoding Met-Ala-(15-

119)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L,

60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 112Q, 116V) SEQ ID NO:357;

DNA Sequence #B27: pMON13378 (Example 96), Met-Asp-(15-119)hIL-3 (18I,

23A, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G,

76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V) SEQ ID NO:358;

I

DNA Sequence #B28: pMON13379 (Example 97), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

112Q, 116V, 120Q and 123E) SEQ ID NO:359;

DNA Sequence #B29: pMON13385 (Example 98), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29V, 32R, 34S, 37P, 42A, 45V, 51R, 55L, 60S, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:360;

DNA Sequence #B30: pMON13381 (Example 99), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82W, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q

and 123E) SEQ ID NO:361;

DNA Sequence #B31: pMON13383 (Example 100), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L,

60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 112Q, 116V, 120Q and 123E) SEQ ID NO:362;

DNA Sequence #B32: pMON13384 (Example 101), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 112Q, 116V,

120Q and 123E) SEQ ID NO:363;

DNA Sequence #B33: pMON13388 (Example 102), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 50D, 51R, 55L, 56S, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:346;

DNA Sequence #B34: pMON13389 (Example 103), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q

and 123E) SEQ ID NO:365;

DNA Sequence #B35: pMON13391 (Example 104), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 34S, 37P, 42A, 45V, 51R, 55L, 60S, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:366;

DNA Sequence #B36: pMON13392 (Example 105), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q

and 123E) SEQ ID NO:367;

DNA Sequence #B37: pMON13393 (Example 106), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45M, 46S, 51R, 55L,

60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:368;

DNA Sequence #B38: pMON13394 (Example 107), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 46S, 51R, 55L, 60S, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:369;

DNA Sequence #B39: pMON13395 (Example 108), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29V, 32R, 34S, 37P, 42D, 45V, 46S, 51R, 55L,

60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:370;

DNA Sequence #B40: pMON13396 (Example 109), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 100R, 101M,

116V, 120Q and 123E) SEQ ID NO:371;

DNA Sequence #B41: pMON13397 (Example 110), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82W, 87S, 93S, 98I, 101A, 105Q, 109E, 100R, 101M,

116V, 120Q and 123E) SEQ ID NO:372;

DNA Sequence #B42: pMON13398 (Example 111), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 51R, 55L, 60S, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:373;

DNA Sequence #B43: pMON13399 (Example 112), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29V, 32R, 34S, 37P, 42D, 45V, 46S, 51R, 55L,

60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:374;

DNA Sequence #B44: pMON13404 (Example 113), encoding Met-Ala-(15-

119)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 112Q, 116V)

SEQ ID NO:375;

DNA Sequence #B45: pMON13387 (Example 114), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 50D, 51R, 55L, 60S, 62V,

67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,

120Q and 123E) SEQ ID NO:376;

DNA Sequence #B46: pMON13416 (Example 115), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45V, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:377;

DNA Sequence #B47: pMON13417 (Example 116), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42D, 45M, 46S, 50D, 51R, 55L, 60S,

62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:378;

DNA Sequence #B48: pMON13420 (Example 117), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45V, 46S, 50D, 51R,

55L, 56S, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:379;

DNA Sequence #B49: pMON13421 (Example 118), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45M, 46S, 50D, 51R,

55L, 56S, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:380;

DNA Sequence #B50: pMON13432 (Example 119), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29R, 32A, 34S, 37P, 42D, 45M, 46S, 50D, 51R,

55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

1O5Q, 109E, 116V, 120Q and 123E) SEQ ID NO:381;

DNA Sequence #B51: pMON13382 (Example 120), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 112Q, 116W,

120Q and 123E) SEQ ID NO:382;

DNA Sequence #B52: pMON13476 (Example 85), Met-Asp-(15-125)hIL-3 (18I,

23A, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G,

76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E)

SEQ ID NO:383;

DNA Sequence #B53: pMON13446 (Example 121), encoding Met-Ala-Tyr-Pro-

Glu-Thr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-(15-125)hIL-3 (18I, 25H, 29R,

32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q,

87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:404;

DNA Sequence #B54: pMON13390 (Example 122), encoding Met-Ala-Tyr-Pro-

Glu-Thr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-(15-125)hIL-3 (18I, 25H, 29R,

32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q,

87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:405;

DNA Sequence #C-1: pMON13418 (Example 123), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76P, 793, 82Q, 85V, 87Y, 88W, 91P, 93S, 95T, 98T, 101A,

105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:393;

DNA Sequence #C-2: pMON13400 (Example 124), encoding Met-Ala-(15-

125)hIL-3 (18I, 20P, 23A, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S,

51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:384;

DNA Sequence #C-3: pMON13402 (Example 125), encoding Met-Ala-(15-

125)hIL-3 (18I, 23L, 25H, 291, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R,

55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:385;

DNA Sequence #C-4: pMON13403 (Example 126), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 50D, 51S, 55T, 59L, 62P,

63H, 67Q, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E,

116V, 120Q and 123E) SEQ ID NO:388;

DNA Sequence #C-5: pMON13411 (Example 127), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 981, 101A, 105Q, 109L, 112Q, 116S,

120Q and 123E) SEQ ID NO:390;

DNA Sequence #C-6: pMON13412 (Example 128), encoding Met-Ala-(15-

118)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109L, 112Q, 1165)

SEQ ID NO:391;

DNA Sequence #C-7: pMON13413 (Example 129), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q

and 123E) SEQ ID NO:392;

DNA Sequence #C-8: pMON13419 (Example 126), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 50D, 515, 55T, 59L, 62P,

63H, 65S, 67Q, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 981, 101A, 105Q,

109E, 116V, 120Q and 123E) SEQ ID NO:389;

DNA Sequence #C-9: pMON13428 (Example 133), encoding Met-Ala-(15-

125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H,

69E, 73G, 76P, 79S, 82Q, 85V, 87Y, 91P, 93S, 95T, 98T, 101A, 105Q,

109L, 112Q, 116S, 120Q and 123E) SEQ ID NO:394;

DNA Sequence #C-10: pMON13440 (Example 131), encoding Met-Ala-(15-

125)hIL-3 (18I, 23A, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R,

55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

1O5Q, 109E, 116V, 120Q and 123E) SEQ ID NO:386;

DNA Sequence #C-11: pM0N13451 (Example 132), encoding Met-Ala-(15-

125)hIL-3 (18I, 19I, 20L, 23A, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V,

46S, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I,

101A, 105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:387;

DNA Sequence #C-12: pMON13459 (Example 134), encoding Met-Ala-(15-

125)hIL-3 (18I, 23L, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R,

55T, 59L, 62V, 67H, 69E, 73G, 76P, 795, 82Q, 85V, 87Y, 91P, 93S, 95T,

98T, 101A, 105Q, 109L, 112Q, 116S, 120Q and 123E) SEQ ID NO:395;

DNA Sequence #C-13: pMON13467 (Example 135), encoding Met-Ala-(15-

125)hIL-3 (181, 23L, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R,

55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,

105Q, 109L, 112Q, 116S, 120Q and 123E) SEQ ID NO:396;

DNA Sequence #C-14: pMON13492 (Example 136), encoding Met-Ala-(15-

125)hIL-3 (18I, 23L, 25H, 29I, 32N, 34S, 37S, 38A, 42S, 45V, 46S, 51R,

55T, 59L, 62V, 67H, 69E, 73G, 76P, 79S, 82Q, 85V, 87Y, 91P, 93S, 95T,

98T, 101A, 105Q, 109E, 116V, 120Q and 123E) SEQ ID NO:397.

Polypeptides corresponding to [SEQ ID NO. 129] comprising (1-133)hIL-3 containing four of more amino acid substitutions can be made using the procedures described above and in the following examples by starting with the appropriate oligonuctiotides and then constructing the DNA encoding the polypeptide and expressing it in an appropriate host cell. In a similar manner polypeptides which correspond to [SEQ ID NO. 130] and contain four or more amino acid substitutions and wherein from 1 to 14 amino have been sequentially deleted from the N-terminus, or from 1 to 15 amino acids have been deleted from the C-terminus or deletions of amino acids have been made from both the N-terminus and the C-terminus can also be made by following the procedures described above and in the following examples, beginning with the appropriate starting materials.

Further details known to those skilled in the art may be found in T. Maniatis, et al.,

Molecular Cloning, A Laboratory Manual,

Cold Spring Harbor Laboratory (1982) and references cited therein, incorporated herein by references; and in J. Sambrook, et al.,

Molecular Cloning, A Laboratory Manual,

2nd edition, Cold Spring Harbor Laboratory (1989) and references cited therein, incorporated herein by reference.

The following examples will illustrate the invention in greater detail although it will be understood that the invention is not limited to these specific examples.

Amino acids are shown herein by standard one letter or three letter abbreviations as follows:

Abbreviated Designation

Amino Acid

A

Ala

Alanine

C

Cys

Cysteine

D

Asp

Aspartic acid

E

Glu

Glutamic acid

F

Phe

Phenylalanine

G

Gly

Glycine

H

His

Histidine

I

Ile

Isoleucine

K

Lys

Lysine

L

Leu

Leucine

M

Met

Methionine

N

Asn

Asparagine

P

Pro

Proline

Q

Gln

Glutamine

R

Arg

Arginine

S

Ser

Serine

T

Thr

Threonine

V

Val

Valine

W

Trp

Tryptophan

Y

Tyr

Tyrosine

Various other examples will be apparent to the person skilled in the art after reading the present disclosure without departing from the spirit and scope of the invention. It is intended that all such other examples be included within the scope of the appended claims.

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J. Immunol.

138:4042 (1987).

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34:315-323 (1985).

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Escherichia coli. Gene,

68: 193-203 (1988).

Yanisch-Perron, C., J. Viera and J. Messing. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.

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33: 103-119 (1985).

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3: 479 (1984).

EXAMPLE 1

Construction of pMON 5846 (

FIG. 4

) which Encodes [Met-(1-133) hIL-3 (Arg

129

)]

A plasmid containing the gene for the cDNA of hIL-3 cloned into pUC18 on an EcoRI to HindIII fragment was obtained from British Biotechnology Limited (Cambridge, England). This plasmid was designated pPO518. The purified plasmid DNA was cleaved by the restriction endonucleases NheI and BamHI. Approximately 0.5 micrograms of cleaved plasmid DNA was ligated to 1.0 picomoles of a pair of annealed oligonucleotides with the following sequence:

5′-CTAGCGATCTTTTAATAAGCTTG-3′

[SEQ ID NO: 1]

3′-GCTAGAAAATTATTCGAACCTAG-5′

[SEQ ID NO: 2]

The ligation mixture was used to transform competent JM101 cells to ampicillin resistance. Colonies were picked, and plasmid DNA was purified and subjected to restriction enzyme analysis. An isolate was identified in which the above oligonucleotide sequence had replaced the portion of the gene that encodes the extreme C terminus. Within the new sequence was a new stop codon, TAA, and a recognition site for the enzyme HindIII. The new plasmid was designated pMON5846.

EXAMPLE 2

(a) Construction of Expression Vector Plasmid pMON2341

The plasmid pMON2341 was used to supply the particular replicon and expression elements used for construction of many of the plasmids used to produce hIL-3 and hIL-3 muteins in

E. coli.

These expression elements are described in the materials and methods section. pMON2341 is derived from pMON5515 (Olins et al., 1988) and from pMON2429. pMON2429 consists of the phage mp18 (Yanisch-Perron et al., 1985) with a BclI fragment carrying the chloramphenicol acetyl transferase (cat) gene from pBR328 (Covarrubias et al., 1981) inserted into the BamHI site. The cat gene in pMON2429 has been altered from that in pBR328 by site directed mutagenesis (Kunkel, 1985). The recognition sites for NcoI and EcoRI which occur in the native gene were altered so that these two restriction enzymes no longer recognize these sites. The changes did not alter the protein specified by the gene. Also, an NcoI site was introduced at the N-terminus of the coding sequence so that it overlaps the codon for initiator methionine.

The steps involved in construction of pMON2341 are listed below:

(1) The DNAs of pMON5515 and pMON2429 were treated with NcoI and HindIII. The fragments were ligated and used to transform competent

E. coli

to ampicillin resistance. From these colonies, some were identified that were chloramphenicol resistant. From one of these colonies, plasmid DNA was isolated in which the rat atriopeptigen gene of pMON5515 had been replaced by the NcoI to HindIII fragment containing the cat gene from pMON2429. This fragment contains the recognition sites for several restriction enzymes in the portion derived from the multilinker region of mp18. The new plasmid was designated pMON2412.

(2) pMON2412 was treated with the enzyme ClaI which cleaves at one location in the pBR327 derived portion of the DNA. The protruding ends were rendered blunt by treatment with Klenow in the presence of nucleotide precursors. This DNA was mixed with an isolated 514 bp RsaI fragment derived from pEMBL8 (Dente et al., 1983). This RsaI fragment contains the origin of replication of phage f1. This ligation mixture was used to transform competent

E. coli

cells to ampicillin resistance. Among the plasmid DNAs isolated from these cells was pMON5578. This plasmid has the structure of pMON2412 with the f1 origin region inserted into the ClaI site. This is illustrated in the Figures and in Olins and Rangwala (1990).

(3) The DNA of pMON5578 was treated with restriction enzymes HindIII and MstII. The DNA was then treated with Klenow enzyme in the presence of nucleotide precursors to render the ends blunt. This treated DNA was ligated and used to transform competent

E. coli

to ampicillin resistance. From the ampicillin resistant colonies, one plasmid was recovered from which the portion between HindIII and MstII was absent. This deletion resulted in the removal of sequences from the plasmid which are recognized by a number of restriction endonuclease sites. The new plasmid was designated pMON5582.

(4) The DNA of pMON5582 was treated with SstII and BclII and ligated in the presence of annealed oligonucleotides with the sequences shown below.

5′- GGCAACAATTTCTACAAAACACTTGATACTGTATGAGCAT-

3′-CGCCGTTGTTAAAGATGTTTTGTGAACTATGACATACTCGTA-

ACAGTATAATTGCTTCAACAGAACAGATC-3′ [SEQ ID NO:3]

TGTCATATTAACGAAGTTGTCTTGT-5′ [SEQ ID NO:4]

This sequence encodes the essential elements of the recA promoter of

E. coli

including the transcription start site and the lexA repressor binding site (the operator) (Sancar et al., 1980). The plasmid recovered from the ligation mixes contained this recA promoter in place of the one in pMON5582 (and in pMON5515). The functionality of the recA promoter was illustrated by Olins and Rangwala (1990). The new plasmid was designated pMON5594.

(5) To eliminate the single EcoRI site in pMON5594, the DNA was treated with EcoRI, then with Klenow in the presence of nucleotide precursors to render the ends blunt and then the DNA was ligated. From this ligation mix a plasmid was recovered whose DNA was not cleaved with EcoRI. This plasmid was designated pMON5630.

(6) To alter the single recognition site for PstI, plasmid pMON5630 was subjected to site directed mutagensis (Kunkel, 1985). The oligonucleotide used in this procedure has the sequence shown below.

5′-CCATTGCTGCCGGCATCGTGGTC-3′ [SEQ ID NO:5]

The result of the procedure was to construct pMON2341 which differs from pMON5630 in that the PstI site in the beta-lactamase gene was altered so that PstI no longer recognizes the site. The single nucleotide change does not alter the amino acid sequence of the beta-lactamase protein.

(b) Construction of pMON5847 (

FIG. 5

) which Encodes [Met-(1-133)hIL-3 (Arg

129

)]

Plasmid pMON2341 was used to supply the replicon, promotor, ribosome binding site, transcription terminator and antibiotic resistance marker for the plasmids used to produce hIL-3 in

E. coli

from cDNA derived hIL-3 genes.

Plasmid pMON2341 was treated with restriction enzymes NcoI and HindIII. The restriction fragment containing the replication origin was purified. The DNA of plasmid pMON5846 was treated with NcoI and HindIII. The restriction fragment containing the hIL-3 gene was gel purified. These purified restriction fragments were mixed and ligated. The ligation mixture was used to transform competent JM101 cells to ampicillin resistance. Colonies were picked, and plasmid DNA was purified and analyzed using restriction enzymes. pMON5847 was identified as a plasmid with the replicon of pMON2341 and the hIL-3 gene in place of the chloramphenicol acetyl transferase gene. JM101 cells harboring this plasmid were cultured in M9 medium and treated with nalidixic acid as described above. Samples of the culture were examined for protein content. It was found that this hIL-3 mutein was produced at about 6% of total cell protein as measured on Coomassie stained polyacrylamide gels.

EXAMPLE 3

Construction of pMON5854 (

FIG. 7

) which Encodes [Met-(1-133)hTL-3 (Arg

129

)]

To increase the accumulation of hIL-3 in

E. coli,

the coding sequence of the amino terminal portion of the protein was altered to more closely reflect the codon bias found in

E. coli

genes that produce high levels of proteins (Gouy and Gautier, 1982). To change the coding sequence for the amino terminal portion of the gene, a pair of synthetic oligonucleotides were inserted between the NcoI and HpaI sites within the coding sequence. About 0.5 micrograms of DNA of the plasmid pMON5847 (Example 2) was treated with NcoI and HpaI. This DNA was mixed with an annealed pair of oligonucleotides with the following sequence:

5′-CATGGCTCCAATGACTCAGACTACTTCTCTTAAGACT-

3′-CGAGGTTACTGAGTCTGATGAAGAGAATTCTGA-

TCTTGGGTT-3′ [SEQ ID NO:6]

AGAACCCAA-5′ [SEQ ID NO:7]

The fragments were ligated. The ligation mixture was used to transform competent JM101 to ampicillin resistance. Colonies were picked into broth. From the cultures plasmid DNA was made and examined for the presence of a DdeI site (CTNAG) which occurs in the synthetic sequence but not between the NcoI and HpaI sites in the sequence of pMON5847. The new recombinant plasmid was designated pMON5854. The nucleotide sequence of the DNA in the coding sequence of the amino terminal portion of the hIL-3 gene in pMON5854 was determined by DNA sequencing and found to be the same as that of the synthetic oligonucleotide used in ligation. Cultures of JM101 cells harboring this plasmid were grown and treated with nalidixic acid to induce production of the hIL-3 mutant protein. Analysis of the proteins on Coomassie gels showed that the accumulation of hIL-3 mutein was about 25% of total cell protein in cultures harboring pMON5854, significantly higher than it was in cultures harboring pMON5847.

EXAMPLE 4

Construction of pMON5887 (

FIG. 12

) which Encodes [Met-(1-125)hIL-3]

The plasmid DNA of pMON5854 (Example 3) was treated with EcoRI and HindIII and the larger fragment gel was purified. About 0.5 microgram of this DNA was ligated to 1 picomole of an annealed pair of oligonucleotides which encode amino acids 107 through 125 of hIL-3. The sequences of these oligonucleotides are shown below.

EcoRI to HindIII

5′-AATTCCGTCGTAAACTGACCTTCTATCTGAAAA-

3′-GGCAGCATTTGACTGGAAGATAGACTTTT-

[SEQ ID NO:8]

CCTTGGAGAACGCGCAGGCTCAACAGTAATA-3′

[SEQ ID NO:9]

GGAACCTCTTGCGCGTCCGAGTTGTCATTATTCGA-5′

After ligation, the DNA was used to transform competent JM101 cells to ampicillin resistance. Colonies were picked into broth and plasmid DNA was isolated from each culture. Restriction analysis of the plasmid DNA showed the presence of an EcoRI to HindIII fragment smaller than that of pMON5854. The nucleotide sequence of the portion of the coding sequence between the EcoRI and HindIII sites was determined to confirm the accuracy of the replaced sequence. The new plasmid was designated pMON5887 encoding Met-(1-125)hIL-3 which has the following amino acid sequence:

[SEQ ID NO:10]

Met Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr

Ser Trp Val Asn Cys Ser Asn Met Ile Asp Glu Ile

Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu

Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile

Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu

Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala

Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro

Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His

Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe

Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu

Asn Ala Gln Ala Gln Gln

EXAMPLE 5

Construction of pMON5967 which Encodes [Met-Ala-(15-125) hIL-3]

Plasmid DNA of pMON5887 isolated from

E. coli

GM48 (dam-) was cleaved with NcoI and ClaI and ligated to 1 picomole of an annealed pair of oligonucleotides, encoding amino acids [Met Ala (15-20)hIL-3]. The sequence of these oligonucleotides is shown below.

5′-CATGGCTAACTGCTCTAACATGAT-3′ [SEQ ID NO:11]

3′-CGATTGACGAGATTGTACTAGC-5′ [SEQ ID NO:12]

The resulting ligation mix was used to transform competent

E. coli

JM101 cells to ampicillin resistant colonies. Plasmid DNA was isolated from these cells and the size of the inserted fragment was determined to be smaller than that of pMON5887 by restriction analysis using NcoI and NsiI. The nucleotide sequence of the region between NcoI and ClaI was determined and found to be that of the synthetic oligonucleotides. The new plasmid was designated pMON5967 and cells containing it were induced for protein production. Sonicated cell pellets and supernatants were used for protein purification and bio-assay.

EXAMPLE 6

Construction of pMON5978 which Encodes [Met-Ala-(15-125)hIL-3]

Plasmid DNA of pMON5967 isolated from

E. coli

GM48(dam-) was cleaved with ClaI and NsiI and ligated to 1 picomole of an annealed assembly of six oligonucleotides encoding hIL-3 amino acids 20-70 (FIG.

2

). This synthetic fragment encodes three unique restriction sites, EcoRV, XhoI and PstI. The sequence of these oligonucleotides is shown in FIG.

2

.

The resulting ligation mix was used to transform competent

E. coli

JM101 cells to ampicillin resistant colonies. Plasmid DNA was isolated and screened with XbaI and EcoRV for the presence of the new restriction site EcoRV. The DNA sequence of the region between ClaI and NsiI was determined and found to be the same as that of the synthetic oligonucleotides. The new plasmid was designated pMON5978, and cells containing it were induced for protein production. Sonicated cell pellets and supernatants were used for protein purification and bio-assay.

Plasmid pMON5978 encodes [Met-Ala-(15-125)hIL-3] which has the following amino acid sequence:

[SEQ ID NO:13]

Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

EXAMPLE 7

Construction of pMON13356

Plasmid pMON5988 DNA was digested with restriction enzymes NcoI and EcoRV, and the resulting 4190 base pair NcoI,EcoRV fragment contains the following genetic elements: beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 47-125 of (15-125)hIL-3. The 4190 base pair NcoI,EcoRV restriction fragment from pMON5988 was ligated to the following annealed complementary oligonucleotides from Table (2).

Oligo #13 [SEQ ID NO:27]

Oligo #14 [SEQ ID NO:28]

The ligation reaction mixture was used to transform

E. coli

K-12 strain JM101 and transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. In the resulting plasmid the 99 bases between the NcoI and EcoRV restriction sites in the (15-125) hIL-3 gene are replaced with 22 bases from the above mentioned oligonucleotides. This linker also contains a NdeI recognition sequence.

EXAMPLE 8

Construction of pMON13344

Plasmid pMON13356 DNA was digested with restriction enzymes NcoI and EcoRV, and the resulting 4190 base pair NcoI,EcoRV fragment contains the following genetic elements: beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 47-125 of (15-125)hIL-3. The second DNA fragment was generated by synthetic gene assembly using the following complementary oligonucleotide pairs that have overlapping ends:

Oligo #1 [SEQ ID NO:15]

Oligo #2 [SEQ ID NO:16]

Oligo #3 [SEQ ID NO:17]

Oligo #4 [SEQ ID NO:18]

Oligo #9 [SEQ ID NO:23]

Oligo #10 [SEQ ID NO:24]

The assembled oligonucleotides create NcoI and EcoRV restriction ends and the DNA sequence that encodes amino acids 15-46 of (15-125)hIL-3 with the following amino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A and 45V. The codons encoding amino acids 15-46 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those, positions where amino acid substitutions were made. The 4190 base pair NcoI,EcoRV restriction fragment from pMON13356 was ligated with the pairs of annealed oligonucleotides. The ligation reaction was digested with NdeI and subsequently used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA sequence was determined to be that of the oligonucleotides. The plasmid, pMON13344, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #2

Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:66]

Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala

Glu Asp Val Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn

Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu

Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly

Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr

Leu Glu Asn Ala Gln Ala Gln Gln

DNA sequence #10 [SEQ ID NO:106) codes for the foregoing pMON13344 polypeptide.

EXAMPLE 9

Construction of pMON13345

The 4190 base pair NcoI,EcoRV restriction fragment from pMON13356 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #1 [SEQ ID NO:15]

Oligo #2 [SEQ ID NO:16]

Oligo #5 [SEQ ID NO:19]

Oligo #6 [SEQ ID NO:20]

Oligo #11 [SEQ ID NO:25]

Oligo #12 [SEQ ID NO:26]

The assembled oligonucleotides create NcoI and EcoRV restriction ends and the DNA sequence that encodes amino acids 15-46 of (15-125)hIL-3 with the following amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S and 45M. The codons encoding amino acids 15-46 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13345, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #3

[SEQ ID NO:67]

Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Asn Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #11 [SEQ ID NO:107] codes for the foregoing pMON13345 polypeptide.

EXAMPLE 10

Construction of pMON13346

The 4190 base pair NcoI,EcoRV restriction fragment from pMON13356 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #1 [SEQ ID NO:15]

Oligo #2 [SEQ ID NO:16]

Oligo #7 [SEQ ID NO:21]

Oligo #8 [SEQ ID NO:22]

Oligo #11 [SEQ ID NO:25]

Oligo #12 [SEQ ID NO:26]

The assembled oligonucleotides create NcoI and EcoRV restriction ends and the DNA sequence that encodes amino acids 15-46 of (15-125)hIL-3 with the following amino acid substitutions: 18I, 25H, 29V, 32A, 37S, 42S and 45M. The codons encoding amino acids 15-46 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth and DNA sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13346, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #4

[SEQ ID NO:68]

Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Val Pro Pro Ala Pro Leu Leu Asp

Ser Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #12 (SEQ ID NO:108] codes for the foregoing pMON13346 polypeptide.

EXAMPLE 11

Construction of pMON13357

Plasmid pMON5988 DNA was digested with restriction enzymes EcoRV and NsiI, and the resulting 4218 base pair EcoRV,NsiI fragment contains the following genetic elements: beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-46 and 72-125 of (15-125)hIL-3.

The 4218 base pair EcoRV,NsiI restriction fragment from pMON5988 was ligated to the following annealed complementary oligonucleotides:

Oligo #19 [SEQ ID NO:33]

Oligo #20 [SEQ ID NO:34]

The ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth, and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. In the resulting plasmid the 71 bases between the EcoRV and NsiI restriction sites in the (15-125)hIL-3 gene are replaced with 22 bases from the above mentioned oligonucleotides. This linker also contains a NdeI recognition sequence.

EXAMPLE 12

Construction of pMON13347

The 4218 base pair EcoRV,NsiI restriction fragment from pMON13357 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #21 [SEQ ID NO:35]

Oligo #22 [SEQ ID NO:36]

Oligo #25 [SEQ ID NO:39]

Oligo #26 [SEQ ID NO:40]

Oligo #31 [SEQ ID NO:45]

Oligo #32 [SEQ ID NO:46]

The assembled oligonucleotides create EcoRV and NsiI restriction ends and the DNA sequence that encodes amino acids 47-71 of (15-125)hIL-3 with the following amino acid substitutions: 51R, 55L, 59L, 62V, 67N and 69E. The codons encoding amino acids 47-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13347, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #5

[SEQ ID NO:69]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #13 [SEQ ID NO:109] codes for the foregoing pMON13347 polypeptide.

EXAMPLE 13

Construction of pMON-13348

The 4218 base pair EcoRV,NsiI restriction fragment from pMON13357 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #21 [SEQ ID NO:35]

Oligo #22 [SEQ ID NO:36]

Oligo #27 [SEQ ID NO:41]

Oligo #28 [SEQ ID NO:42]

Oligo #31 [SEQ ID NO:45]

Oligo #32 [SEQ ID NO:46]

The assembled oligonucleotides create EcoRV and NsiI restriction ends and the DNA sequence that encodes amino acids 47-71 of (15-125)hIL-3 with the following amino acid substitutions: 51R, 55L, 60S, 62V, 67N and 69E. The codons encoding amino acids 47-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13348, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #6

[SEQ ID NO:70]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #14 [SEQ ID NO:110] encodes the foregoing pMON13348 polypeptide.

EXAMPLE 14

Construction of pMON13349

The 4218 base pair EcoRV,NsiI restriction fragment from pMON13357 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #23 [SEQ ID NO:37]

Oligo #24 [SEQ ID NO:38]

Oligo #25 [SEQ ID NO:39]

Oligo #26 [SEQ ID NO:40]

Oligo #29 [SEQ ID NO:43]

Oligo #30 [SEQ ID NO:44]

The assembled oligonucleotides create EcoRV and NsiI restriction ends and the DNA sequence that encodes amino acids 47-71 of (15-125)hIL-3 with the following amino acid substitutions: 51R, 55T, 59L, 62V, 67H and 69E. The codons encoding amino acids 47-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth and the DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13349, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #7

[SEQ ID NO:71]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #15 [SEQ ID NO:111] encodes the foregoing pMON13349 polypeptide.

EXAMPLE 15

Construction of pMON13358

Plasmid pMON5988 DNA was digested with restriction enzymes NsiI and EcoRI and the resulting 4178 base pair NsiI,EcoRI fragment contains the following genetic elements: beta-latamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-71 and 106-125 of (15-125)hIL-3. The 4178 base pair NsiI,EcoRI restriction fragment from pMON5988 was ligated to the following annealed complementary oligonucleotides.

Oligo #15 [SEQ ID NO:29]

Oligo #16 [SEQ ID NO:30]

The ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth, and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. In the resulting plasmid the 111 bases between the NsiI and EcoRI restriction sites in the (15-125) hIL-3 gene are replaced with 24 bases from the above mentioned oligonucleotides. This linker also contains a NdeI recognition sequence.

EXAMPLE 16

Construction of pMON13350

The 4178 base pair NsiI,EcoRI restriction fragment from pMON13358 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #41 [SEQ ID NO:55]

Oligo #42 [SEQ ID NO:56]

Oligo #39 [SEQ ID NO:53]

Oligo #40 [SEQ ID NO:54]

Oligo #35 [SEQ ID NO:49]

Oligo #36 [SEQ ID NO:50]

Oligo #43 [SEQ ID NO:57]

Oligo #44 [SEQ ID NO:58]

The assembled oligonucleotides create NsiI and EcoRI restriction ends and the DNA sequence that encodes amino acids 72-105 of (15-125)hIL-3 with the following amino acid substitutions: 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A and 105Q. The codons encoding amino acids 72-105 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13350, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #8

[SEQ ID NO:72]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #16 [SEQ ID NO:112] codes for the foregoing pMON13350 polypeptide.

EXAMPLE 17

Construction of pMON13355

The 4178 base pair NsiI,EcoRI restriction fragment from pMON13358 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #41 [SEQ ID NO:55]

Oligo #42 [SEQ ID NO:56]

Oligo #37 [SEQ ID NO:51]

Oligo #38 [SEQ ID NO:52]

Oligo #33 [SEQ ID NO:47]

Oligo #34 [SEQ ID NO:48]

Oligo #43 [SEQ ID NO:57]

Oligo #44 [SEQ ID NO:58]

The assembled oligonucleotides create NsiI and EcoRI restriction ends and the DNA sequence that encodes amino acids 72-105 of (15-125)hIL-3 with the following amino acid substitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A and 1050. The codons encoding amino acids 72-105 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13355, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #9

[SEQ ID NO:73]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #17 [SEQ ID NO:113] codes for the foregoing pMON13355 polypeptide.

EXAMPLE 18

Construction of pMON13359

Plasmid pMON5988 DNA was digested with restriction enzymes EcoRI and HindIII, and the resulting 4225 base pair EcoRI,HindIII fragment contains the following genetic elements: beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-105 of (15-125)hIL-3. The 4225 base pair EcoRI,HindIII restriction fragment from pMON5988 was ligated to the following annealed complementary oligonucleotides.

Oligo #17 [SEQ ID NO:31]

Oligo #18 [SEQ ID NO:32]

The ligation reaction was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth, and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. In the resulting plasmid the 64 bases between the EcoRI and HindIII restriction sites in the (15-125)hIL-3 gene are replaced with 20 bases from the above mentioned oligonucleotides. This linker also contains an NdeI recognition sequence.

EXAMPLE 19

Construction of pMON13352

The 4225 base pair EcoRI,HindIII restriction fragment from pMON13359 was ligated with the following pairs of annealed complementary oligonucleotides:

Oligo #45 [SEQ ID NO:59]

Oligo #46 [SEQ ID NO:60]

Oligo #49 [SEQ ID NO:63]

Oligo #50 [SEQ ID NO:64]

The assembled oligonucleotides create EcoRI and HindIII restriction ends and the DNA sequence that encodes amino acids 106-125 of (15-125)hIL-3 with the following amino acid substitutions: 109E, 116V, 120Q and 123E. The codons encoding amino acids 106-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth. The DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13352, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #10

[SEQ ID NO:74]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence #18 [SEQ ID NO:114] codes for the foregoing pMON13352 polypeptide.

EXAMPLE 20

Construction of pMON13354

The 4225 base pair EcoRI,HindIII restriction fragment from pMON13359 was ligated with the following pairs of annealed complementary oligonucleotides:.

Oligo #45 [SEQ ID NO:59]

Oligo #46 [SEQ ID NO:60]

Oligo #47 [SEQ ID NO:61]

Oligo #48 [SEQ ID NO:62]

The assembled oligonucleotides create EcoRI and HindIII restriction ends and the DNA sequence that encodes amino acids 106-125 of (15-125)hIL-3 with the following amino acid substitutions: 109E, 116V, 117S, 120H and 123E. The codons encoding amino acids 106-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The ligation reaction was digested with NdeI and used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated from a colony grown in LB broth, and the DNA was sequenced to determine that the sequence was that of the oligonucleotides. The plasmid, pMON13354, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #11

[SEQ ID NO:75]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His

Ala Gln Glu Gln Gln

DNA sequence #19 [SEQ ID NO:115] codes for the foregoing pMON13354 polypeptide.

EXAMPLE 21

Construction of pMON13360

Plasmid pMON13352 DNA was digested with restriction enzymes NsiI and EcoRI, resulting in a 4178 base pair NsiI,EcoRI fragment. The genetic elements derived from pMON13352 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-71 and 106-125 of (15-125)hIL-3. Plasmid pMON13350 DNA was digested with NsiI and EcoRI. The resulting 111 base pair NsiI, EcoRI fragment encodes amino acids 72-105 of (15-125)hIL-3. The eluted restriction fragments were concentrated and desalted using Centricon 30 concentrators. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and analyzed by restriction analysis. Clones containing the correct insert lost a XmnI site as compared with pMON13352. Positive clones were identified by the loss of a 615 base pair XmnI fragment. The DNA was sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13360, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #12

[SEQ ID NO:76]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lya Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence #23 [SEQ ID NO:119] encodes the foregoing pMON13360 polypeptide.

EXAMPLE 22

Construction of pMON13361

Plasmid pMON13352 DNA was digested with restriction enzymes NsiI and EcoRI, resulting in a 4178 base pair NsiI,EcoRI fragment. The genetic elements derived from pMON13352 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-71 and 106-125 of (15-125)hIL-3. Plasmid pMON13355 DNA was digested with NsiI and EcoRI. The resulting 111 base pair NsiI, EcoRI fragment encodes amino acids 72-105 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Clones containing the correct insert contained an additional RsaI site which results in a 1200 base pairs RsaI fragment. The DNA was sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13361, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #13

[SEQ ID NO:77]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence #24 [SEQ ID NO:120] codes for the foregoing pMON13361 polypeptide.

EXAMPLE 23

Construction of pMON13362

Plasmid pMON13354 DNA was digested with restriction enzymes NsiI and EcoRI, resulting in a 4178 base pair NsiI,EcoRI fragment. The genetic elements derived from pMON13354 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-71 and 106-125 of (15-125)hIL-3. Plasmid pMON13355 DNA was digested with NsiI and EcoRI. The resulting 111 base pair NsiI, EcoRI fragment encodes amino acids 72-105 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Clones containing the correct insert contained an additional RsaI site which results in a 1200 base pairs RsaI fragment. The DNA was sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13362, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #14

[SEQ ID No:78]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Prc Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His

Ala Gln Glu Gln Gln

DNA sequence #25 [SEQ ID NO:121] codes for the foregoing pMON13362 polypeptide.

EXAMPLE 24

Construction of pMON13363

Plasmid pMON13344 DNA was digested with restriction enzymes NsiI and EcoRV, resulting in a 4218 base pair NsiI,EcoRV fragment. The genetic elements derived from pMON13344 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-46 and 72-125 of (15-125)hIL-3. Plasmid pMON13348 DNA was digested with NsiI and EcoRV. The resulting 71 base pair NsiI, EcoRV fragment encodes amino acids 47-71 of (15-125)hIL-3. The restriction fragments were ligated with T4 ligase, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Clones containing the correct insert contained an additional DdeI site which results in DdeI restriction fragments of 806 and 167 base pairs compared to 973 base pairs in pMON13344. The DNA was sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13363, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #15

[SEQ ID NO:79]

Asn Cys Ser Tle Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Ala Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #20 [SEQ ID NO:116] codes for the foregoing pMON13363 polypeptide.

EXAMPLE 25

Construction of pMON13364

Plasmid pMON13345 DNA was digested with restriction enzymes NsiI and EcoRV, resulting in a 4218 base pair NsiI,EcoRV fragment. The genetic elements derived from pMON13345 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-46 and 72-125 of (15-125)hIL-3. Plasmid pMON13349 DNA was digested with NsiI and EcoRV. The resulting 71 base pair NsiI, EcoRV fragment encodes amino acids 47-71 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Clones containing the correct insert contained an additional DdeI site which results in DdeI restriction fragments of 806 and 167 base pairs compared to 973 base pairs in pMON13344. The DNA was sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13364, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #16

[SEQ ID NO:80]

Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Asn Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #21 [SEQ ID NO:117] codes for the foregoing pMON13364 polypeptide.

EXAMPLE 26

Construction of pMON13365

Plasmid pMON13346 DNA was digested with restriction enzymes NsiI and EcoRV, resulting in a 4218 base pair NsiI,EcoRV fragment. The genetic elements derived from pMON13346 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the bases encoding amino acids 15-46 and 72-125 of (15-125)hIL-3. Plasmid pMON13347 DNA was digested with NsiI and EcoRV. The resulting 71 base pair NsiI, EcoRV fragment encodes amino acids 47-71 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Clones containing the correct insert contained an additional DdeI site which results in DdeI restriction fragments of 806 and 167 base pairs compared to 973 base pairs in pMON13344. The DNA was sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29V, 32A, 37S, 425, 45M, 51R, 55L, 59L, 62V, 67N and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13365, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #17

[SEQ ID NO:81]

Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Val Pro Pro Ala Pro LGu Leu Asp

Ser Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #22 [SEQ ID NO:118] codes for the foreging pMON13365 polypeptide.

EXAMPLE 27

Construction of pMON13298

Plasmid pMON5978 DNA was digested with restriction enzymes NsiI and HindIII, resulting in a 3789 base pair NsiI,HindIII fragment. The genetic elements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site, and the bases encoding amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13360 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13298, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #18

[SEQ ID NO:82]

Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence #29 [SEQ ID NO:125] codes for the foregoing pMON13298 polypeptide.

EXAMPLE 28

Construction of pMON13299

Plasmid pMON5978 DNA was digested with restriction enzymes NsiI and HindIII, resulting in a 3789 base pair NsiI,HindIII fragment. The genetic elements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the bases encoding amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13361 DNA was digested with NsiI and HindIII, the resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116v, 120Q and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13299, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #19

[SEQ ID NO:83]

Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence #30 [SEQ ID NO:126] codes for the foregoing pMON13299 polypeptide.

EXAMPLE 29

Construction of pMON13300

Plasmid pMON5978 DNA was digested with restriction enzymes NsiI and HindIII, resulting in a 3789 base pair NsiI,HindIII fragment. The genetic elements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site, and the bases encoding amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13362 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13300, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #20

[SEQ ID NO:84]

Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His

Ala Gln Glu Gln Gln

DNA sequence #31 [SEQ ID NO:127] codes for the foregoing pMON13300 polypeptide.

EXAMPLE 30

Construction of pMON13301

Plasmid pMON5978 DNA was digested with restriction enzymes NcoI and NsiI, resulting in a 3794 base pair NcoI,NsiI fragment. The genetic elements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the bases encoding amino acids 72-125 of (15-125)hIL-3. Plasmid pMON13363 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13301, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #21

[SEQ ID NO:85]

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Ala Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #26 [SEQ ID NO:122] codes for the foregoing pMON13301 polypeptide.

EXAMPLE 31

Construction of pMON13302

Plasmid pMON5978 DNA was digested with restriction enzymes NcoI and NsiI, resulting in a 3794 base pair NcoI, NsiI fragment. The genetic elements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site, and the bases encoding amino acids 72-125 of (15-125)hIL-3. Plasmid pMON13364 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13302, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #22

[SEQ ID NO:86]

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Asn Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #27 [SEQ ID NO:123] codes for the foregoing pMON13302 polypeptide.

EXAMPLE 32

Construction of pMON13303

Plasmid pMON5978 DNA was digested with restriction enzymes NcoI and NsiI, resulting in a 3794 base pair NcoI,NsiI fragment. The genetic elements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site, and the bases encoding amino acids 72-125 of (15-125)hIL-3. Plasmid pMON13365 DNA was digested with Ncol and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13303, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #23

[SEQ ID NO:87]

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Val Pro Pro Ala Pro Leu Leu Asp

Ser Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His I1e Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

DNA sequence #28 [SEQ ID NO:124] codes for the foregoing pMON13303 polypeptide.

EXAMPLE 33

Construction of pMON13287

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13363 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13360 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13287, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #24

[SEQ ID NO:88]

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Ala Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu

Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser

Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys

Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro

Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence #1 [SEQ ID NO:97] codes for the foregoing pMON13287 polypeptide.

EXAMPLE 34

Construction of pMON13288

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13364 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoiI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13360 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13288, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #25

[SEQ ID NO:89]

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile

His His Leu Lys Arg Pro Pro Asn Pro Leu Leu Asp

Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu

Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala

Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys

Leu Pro Ser Ala Thr Ala Ala Prc Ser Arg His Pro

Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg

Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln

Ala Gln Glu Gln Gln

DNA sequence 44 [SEQ ID NO:100] codes for the foregoing pMON13288 polypeptide.

EXAMPLE 35

Construction of pMON13289

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13365 DNA was digested with NcoI and NsiI. The resulting 170 base pair Ncoi, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13360 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 935, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13289, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #26

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:90]

Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser

Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn

Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser

Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Giy

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr

Leu Glu Gln Ala Gln Glu Gln Gln

DNA sequence #7 [SEQ ID NO:103] codes for the foregoing pMON13289 polypeptide.

EXAMPLE 36

Construction of pMON13290

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13363 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13361 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13290, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #27

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:91]

Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala

Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu PrO Asn

Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser

Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr

Leu Glu Gln Ala Gln Glu Gln Gln

DNA sequence #2 [SEQ ID NO:98] codes for the foregoing pMON13290 polypeptide.

EXAMPLE 37

Construction of pMON13292

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13365 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13361 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13292, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #28

Met Ala Asn cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:92]

Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser

Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn

Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr

Leu Glu Gln Ala Gln Glu Gln Gln

DNA sequence #8 [SEQ ID NO:104] codes for the foregoing pMON13292 polypeptide.

EXAMPLE 38

Construction of pMON13294

Plasmid pMON2341 DNA was digested with restriction enzymes NCoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13364 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13362 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32N, 35 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13294, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #29

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:93]

Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser

Glu Asp Met AsP Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn

Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser

Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser

Leu Glu His Ala Gln Glu Gln Gln

DNA sequence #6 [SEQ ID NO:102] codes for the foregoing pMON13294 polypeptide.

EXAMPLE 39

Construction of pMON13295

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13365 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13362 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13295, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #30

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:94]

Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser

Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn

Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser

Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser

Leu Glu His Ala Gln Glu Gln Gln

DNA sequence #9 [SEQ ID NO:105] codes for the foregoing pMON13295 polypeptide.

EXAMPLE 40

Construction of pMON13312

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13364 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13361 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V, 120Q and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13312, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #31

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:95]

Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser

Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn

Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser

Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr

Leu Glu Gln Ala Gln Glu Gln Gln

DNA sequence #5 [SEQ ID NO:101] codes for the foregoing pMON13312 polypeptide.

EXAMPLE 41

Construction of pMON13313

Plasmid pMON2341 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter and g10L ribosome binding site. Plasmid pMON13363 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON13362 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis, and sequenced to confirm the correct insert. The resulting (15-125)hIL-3 variant has the following amino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 935, 98T, 101A, 105Q, 109E, 116V, 117S, 120H and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13313, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

Peptide #32

Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu

[SEQ ID NO:96]

Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala

Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn

Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser

Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser

Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala Gly

Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser

Leu Glu His Ala Gln Glu Gln Gln

DNA sequence #3 [SEQ ID NO:99] codes for the foregoing pMON13313 polypeptide.

EXAMPLE 42

Construction of pMON5987

Plasmid pMON6458 DNA was digested with restriction enzymes NcoI and HindIII, resulting in a 3940 base pair NcoI,HindIII fragment. The genetic elements derived from pMON6458 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site and lamB secretion leader. Plasmid pMON5978 DNA was digested with NcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125)hIL-3. Plasmid pMON5976 DNA was digested with NsiI and HindIII. The resulting 175 base pair NsiI,HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and screened for the restriction sites EcoRV and NheI and DNA sequenced to confirm the correct insert.

EXAMPLE 43

Construction of pMON5988

The plasmid DNA of pMON5987 was digested with NheI and EcoRI, resulting in a 3903 base pair NheI, EcoRI fragment. The 3903 base pair NheI, EcoRI fragment was ligated to 1.0 picomoles of the following annealed oligonucleotides:

5′-CTAGCCACGGCCGCACCCACGCGACATCCAATCCATATCAA-

3′-GGTGCCGGCGTGGGTGCGCTGTAGGTTAGGTATAGTT-

GGACGGTGACTGGAATG-3′ [SEQ ID NO:131]

CCTGCCACTGACCTTACAATT-5′ [SEQ ID NO:132]

The ligation reaction mixture was used to transform

E. coli

K-12 strain JM101 and transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and sequenced to confirm positive clones. This plasmid was constructed to change alanine 101 to aspartic acid in the hIL-3 gene (15-125). This plasmid was designated pMON5988.

EXAMPLE 44

Construction of pMON5853 (

FIG. 6

) which Encodes [Met-(15-133)hIL-3 (Arg

129

)]

Plasmid DNA of pMON5847 (Example 2) was treated with NcoI. The restriction enzyme was inactivated by heat treatment (65° C. for 10 minutes). The DNA was then treated with large fragment of DNA polymerase I (Klenow) in the presence of all four nucleotide precursors. This produces DNA termini with non-overlapping ends. After 5 minutes at 37° C., the polymerase was inactivated by heat treatment at 65° C. for 10 minutes. The DNA was then treated with HpaI, an enzyme which produces non-overlapping termini. The DNA was ethanol precipitated and ligated. The ligation reaction mixture was used to transform competent JM101 cells to ampicillin resistance. Colonies were picked and plasmid DNA was analyzed by restriction analysis. A plasmid designated pMON5853 was identified as one containing a deletion of the amino terminal 14 codons of the hIL-3 gene. The DNA sequence for the junction of the ribosome binding site to the (15-133) hIL-3 gene was determined to be the following:

5′-AAGGAGATATATCCATGAACTGCTCTAAC-3′[SEQ ID NO:133]

M N C S N [SEQ ID NO:134]

The lower line contains the one letter code for the amino acids specified by the coding sequence of the amino terminus of the 15-133 hIL-3 gene. These are methionine, asparagine, cysteine, serine and asparagine.

When cultures of JM101 cells harboring this plasmid were induced with nalidixic acid, it was found that hIL-3 (15-133) accumulated at levels higher than hIL-3 (pMON5847).

The plasmid, pMON5853, encodes Met-(15-133) hIL-3 (Arg

129

) which has the following amino acid sequence:

[SEQ ID NO:135]

Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr

His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn

Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn

Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala

Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile

Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala

Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp

Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys

Thr Leu Glu Asn Ala Gln Ala Gln Gln Thr Thr Leu Arg

Leu Ala Ile Phe

EXAMPLE 45

Construction of pMON5873 which Encodes [Met-(1-133)hIL-3]

The gene obtained from British Biotechnology, Ltd. specified arginine at codon position 129. The amino acid specified in the native hIL-3 cDNA is serine. To produce a protein with the native sequence at this position, the portion of the coding sequence between the EcoRI site at codons 106 and 107 and the NheI site at codons 129 and 130 was replaced. Plasmid DNA of pMON5854 (Example 3) and pMON5853 (Example 44) were treated with EcoRI and NheI. The larger fragments of each were gel purified. These were ligated to a pair of an annealed oligonucleotides with the following sequences:

5′-AATTCCGTCGTAAACTGACCTTCTATCTGAAAACC-

[SEQ ID NO: 136]

3′-GGCAGCATTTGACTGGAAGATAGACTTTTGG-

TTGGAGAACGCGCAGGCTCAACAGACCACTCTGTCG-3′

AACCTCTTGCGCGTCCGAGTTGTCTGGTGAGACAGCGATC-5′

[SEQ ID NO:137]

The ligation reaction mixtures were used to transform competent JM101 cells to ampicillin resistance. Colonies were picked into broth and grown. Plasmid DNA was isolated and screened for the presence of a new StyI recognition site present in the synthetic DNA and not in pMON5854 and pMON5853. The nucleotide sequence of the gene in the region between EcoRI and NheI was determined and found to be that of the synthetic oligonucleotides. The new plasmids were designated pMON5873 encoding [Met-(1-133)hIL-3] and pMON5872 encoding [Met-(15-133)hIL-3].

The plasmid, pMON5873, encodes Met-(1-133)hIL-3 which has the following amino acid sequence:

[SEQ ID NO:128]

Met Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser

Trp Val Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr

His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn

Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn

Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala

Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile

Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala

Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp

Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys

Thr Leu Glu Asn Ala Gln Ala Gln Gln Thr Thr Leu Ser

Leu Ala Ile Phe

EXAMPLE 46

Construction of pMON6458

Plasmid pMON6525 was digested with restriction enzymes HindIII and SalI and the resulting 3172 base pair fragment was isolated from a 1% agarose gel by interception onto DEAE membrane. The genetic elements derived from pMON6525 are the beta-lactamase gene (AMP), pBR327 origin of replication, and phage f1 origin of replication as the transcription terminator. (The genetic elements derived from plasmid pMON6525 are identical to those in plasmid pMON2341 which could also be used to construct pMON6458.) Plasmid pMON6457 was digested with restriction enzymes HindIII and SalI and the resulting 1117 base pair fragment was isolated by PAGE and crush and soak elution. The genetic elements derived from pMON6457 are the pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and the (15-125) hIL-3 gene. The restriction fragments were ligated and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. Clones containing the hIL-3 gene (encoding amino acids 15-125) contained a 345 base pair NcoI, HindIII restriction fragment. This construct was designated pMON6458. This plasmid was constructed to eliminate an EcoRI restriction site outside the hIL-3 gene coding region in plasmid pMON6457.

EXAMPLE 47

Construction of pMON5976 which Encodes [Met-(15-125)hIL-3 (Ala

101

)]

The plasmid DNA of pMON5941 isolated from the dam-

E. coli

strain GM48 was cleaved with ClaI and NsiI and ligated to 1 picomole of an annealed assembly of six oligonucleotides encoding amino acids 20-70 of hIL-3 (FIG.

2

). This synthetic fragment encodes three unique restriction sites, EcoRV, XhoI and PstI. The sequence of these oligonucleotides is shown in FIG.

2

.

The resulting ligation mix was used to transform competent

E. coli

JM101 cells to ampicillin resistant colonies. Plasmid DNA was isolated and the inserted fragment was determined to have both an EcoRV and NheI site. The nucleotide sequence of the region between ClaI and NsiI was determined and found to be that of the synthetic oligonucleotides. At codons 86-87 of a nucleotide sequence coding for (15-125)hIL-3, an NheI site was introduced. The plasmid with this alteration was designated pMON5941. This plasmid encodes Met-(15-125)hIL-3 which is altered at position 101 by replacement of aspartate by alanine.

Plasmid pMON5976 encodes Met-(15-125)hIL-3(Ala

101

) which has the following amino acid sequence:

[SEQ ID NO:138]

Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Ala Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln

EXAMPLE 48

Construction of pMON5917 which Encodes [Met-(15-8)hIL-3]

The plasmid DNA of pMON5853 was cleaved with NsiI and HindIII and ligated to an annealed pair of oligonucleotides encoding (70-88)hIL-3 with a new NheI endonuclease restriction site at codons 86-87. The sequence of these oligonucleotides is shown in Example 18.

The ligation mixture was used to transform competent

E. coli

JM101 cells, and ampicillin resistant colonies were picked. Plasmid DNA isolated from individual colonies was screened for the presence of the new NheI restriction site. The nucleotide sequence of the substituted portion was determined and found to be that of the synthetic oligonucleotides. The new plasmid was designated pMON5917 encoding Met-(15-88)hIL-3 containing a new NheI site at codons 86-87.

Plasmid pMON5917 encodes Met-(15-88)hIL-3 which has the following amino acid sequence:

[SEQ ID NO:139]

Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala

EXAMPLE 49

Construction of pmon5941 which Encodes [Met-(15-125) hIL-3 Ala

101

]

The plasmid DNA of pMON5917 was cleaved with NheI and HindIII and ligated to two annealed pairs of oligonucleotides which encode amino acids 86-106 and 107-125 of hIL-3. The sequences of these oligonucleotides is shown below.

NheI to EcoRI

5′-CTAGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGCTG-

3′-GGTGCCGGCGTGGGTGCGCTGTAGGTTAGGTATAGTTCCGAC-

GTGACTGGAATG-3′

[SEQ ID NO:140]

CACTGACCTTACTTAA-5′

[SEQ ID NO:141]

EcoRI to HindIII

5′-AATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCA-

3′-GGCAGCATTTGACTGGAAGATAGACTTTTGGAACCTCTTGCGCGT-

GGCTCAACAGTAATA-3′

[SEQ ID NO:142]

CCGAGTTGTCATTATTCGA-5′

[SEQ ID NO:143]

The ligation mixture was used to transform competent

E. coli

JM101 cells to ampicillin resistant colonies. Plasmid DNA was isolated from these cells and the size of the inserted fragment was determined to be larger by restriction analysis with NcoI and HindIII. The Asp to Ala 101 change is encoded on the NheI to EcoRI fragment. The nucleotide sequence of the portion of the coding region between the NheI and HindIII sites was determined and found to be that of the synthetic oligonucleotides. The new plasmid was designated pMON5941.

The plasmid, pMON5941, encodes Met-(15-125)hIL-3(Ala

101

) and contains a new NheI restriction site.

EXAMPLE 50

Construction of pMON6455

Plasmid pMON5905 was digested with restriction enzymes HindIII and NcoI resulting in a 3936 base pair fragment. The genetic elements derived from pMON5905 are the beta-lactamase gene (AMP), pBR327 origin of replication, pAraBAD promoter, g10L ribosome binding site, lamB secretion leader and phage f1 origin of replication as the transcription terminator. The following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, g10L ribosome binding site and phage f1 origin of replication as the transcription terminator, derived from plasmid pMON5905 are identical to these in plasmid pMON5594 which could also be used to construct pMON6455. The AraBAD promoter is identical to that described in pMON6235. The lamB signal peptide sequence used in pMON6455 is that shown in

FIG. 8

fused to hIL-3 (15-125) at the NcoI site. Plasmid pMON5887 was digested with restriction enzymes HindIII and NcoI, resulting in a 384 base pair NcoI, HindIII fragment. The restriction fragments were ligated, and the ligation reaction mixture was used to transform into

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. Positive clones containing the hIL-3 gene (encoding amino acids 1-125) contained a 384 base pair NcoI, HindIII restriction fragment. This construct was designated pMON6455.

EXAMPLE 51

Construction of pMON6456

Plasmid pMON5905 was digested with restriction enzymes HindIII and NcoI resulting in a 3936 base pair fragment. The genetic elements derived from pMON5905 are the beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, pAraBAD promoter, g10L ribosome binding site and the lamB secretion leader. Plasmid pMON5871 was digested with restriction enzymes HindIII and NcoI, resulting in a 330 base pair NcoI, HindIII fragment. The genetic element derived from pMON5871 encompassed the bases encoding the (1-107) hIL-3 gene. The restriction fragments were ligated, and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes NcoI and HindIII in double digest. Clones containing the hIL-3 gene (encoding amino acids 1-107) contained a 330 base pair NcoI, HindIII restriction fragment. This construct was designated pMON6456.

EXAMPLE 52

Construction of pMON6457

Plasmid pMON6455 DNA grown in

E. coli

strain GM48 (dam-)was digested with restriction enzymes NcoI and ClaI, resulting in a 4263 base pair NcoI, ClaI fragment. The restriction fragment was ligated to 1.0 picomoles of annealed oligonucleotides with the following sequence coding for Met Ala 14-20 hIL-3:

5′-CATGGCTAACTGCTCTAACATGAT-3′ [SEQ ID NO:151]

3′-CGATTGACGAGATTGTACTAGC-5′ [SEQ ID NO:152]

The resulting DNA was transformed into

E. coli

K-12 strain JM101 and transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated and the size of the inserted fragment was determined by restriction analysis employing restriction enzymes XbaI and EcoRI in double digest. Positive clones containing the hIL-3 gene (encoding aa 15-125 of hIL-3) contained a 433 base pair XbaI, EcoRI restriction fragment. This construct was designated pMON6457. This plasmid was constructed to delete the first 14 amino acids of hIL-3. The coding sequence of the resulting gene begins as follows:

5′ ATG GCT AAC TGC . . . 3′

[SEQ ID NO:153]

Met Ala Asn Cys . . .

[SEQ ID NO:154]

15

The first two amino acids (Methionine, Alanine) create an NcoI restriction site and a signal peptidase cleavage site between the lamB signal peptide and (15-125) hIL-3. Plasmid pMON6457 encodes (15-125) hIL-3 which has the amino acid sequence designated SEQ ID NO:65.

EXAMPLE 53

Construction of pMON6235

One of the DNA fragments used to create this plasmid was generated by site-directed mutagenesis employing PCR techniques described previously using the following oligonucleotides, Oligo #51 [SEQ ID NO:155] and Oligo #52 [SEQ ID NO:156], were used as primers in this procedure. The template for the PCR reaction was

E. coli

strain W3110 chromosomal DNA, prepared as described in Maniatis (1982). The oligonucleotide primers were designed to amplify the AraBAD promoter (Greenfield et al., 1978). The resulting DNA product was digested with the restriction enzymes SacII and BglII. The reaction mixture was purified as described previously. Plasmid, pMON5594, DNA was digested with SacII and BglII, resulting in a 4416 base pair SacII,BglII restriction fragment which contains the following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, G10L ribosome binding site, phage f1 origin of replication as the transcription terminator and the chloramphenicol acetyl transferase (cat) gene. The 4416 base pair SacII,BglII restriction fragment from pMON5594 was ligated to the PCR-generated SacII, BglII DNA fragment. The ligation mixture was used to transform

E. coli

K-112 strain JM101. Positive clones contained a 323 base pair SacII,BglII fragment and were DNA sequenced to confirm that the SacII,BglII fragment was the AraBAD promoter. This construct was designated pMON6235.

EXAMPLE 54

Construction of pMON5647

Plasmid pMON5585 [prepared as disclosed in EP 0241446 incorporated herein by reference in its entirety] DNA was digested with restriction enzymes NcoI and HindIII resulting in a 3273 base pair NcoI,HindIII fragment. The genetic elements derived from pMON5585 are the pBR327 origin of replication, precA promoter, g10L ribosome binding protein, bovine somatotropin gene (bST), beta-lactamase gene (AMP) and T7 transcription terminator. Plasmid pMON3267 [prepared as disclosed in EP 0241446 incorporated herein by reference in its entirety] DNA was digested with NcoI and HindIII enzymes resulting in a 580 base pair NcoI,HindIII fragment which contains the porcine somatotropin (pST) gene. The restriction fragments were ligated and the ligation reaction mixture was used to transform

E. coli

strain JM101. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis and sequenced to confirm the correct insert.

EXAMPLE 55

Construction of pMON710

Plasmid pMON709 consists of a 1614 base pair AvaI,EcoRI fragment of transposon TN7, containing the streptomycin adenylyltransferase gene (Fling et al., 1985) and a pUC9 linker (XmaI,HindIII) cloned between the HindIII and EcoRI sites of pUC19. The streptomycin adenylyltransferase gene COnfers resistance to streptomycin and spectinomycin. Plasmid pMON709 was mutagenized by oligonucleotide site-directed mutagenesis (methods described in Zoller and Smith, 1982) to introduce an EcoRV site at the 3′ end of the streptomycin adenylyltransferase gene. The oligonucleotide, Oligo # 53 [SEQ ID NO:157], was used in this procedure to introduce the EcoRV site. The resulting plasmid was designated pMON710.

EXAMPLE 56

Construction of pMON5723

Plasmid pMON5647 DNA was digested with restriction enzymes DraI and SspI resulting in a 2916 base pair DraI, SspI fragment. The genetic elements derived from pMON5647 are the pBR327 origin of replication, precA promoter, g10L ribosome binding protein, porcine somatotropin gene (pST) and T7 transcription terminator (Dunn and Strudier, 1983). Plasmid pMON710 DNA was digested with restriction enzymes HincII and EcoRV resulting in 940 base pair HincII,EcoRV fragment containing the streptomycin adenylyltransferase gene which infers resistance to streptomycin and spectinomycin. The restriction fragments were ligated and the ligation reaction mixture was used to transform

E. coli

strain JM101. The DraI, SspI, HincII and EcoRV restriction sites are lost as a result of the cloning. Transformant bacteria were selected on spectinomycin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis and sequenced to confirm the correct insert.

EXAMPLE 57

Construction of pMON13361

Plasmid pMON13288 was mutagenized by oligonucleotide site-directed mutagenesis (method described in Kunkel, 1985) to eliminate a NsiI site in the (15-125) hIL-3 variant coding region. Codon 70 of (15-125) hIL-3, encoding asparagine, was converted from AAT to AAC destroying the NsiI recognition site. The oligonucleotide, Oligo # 54 [SEQ ID NO:158], was used in this procedure to eliminate the NsiI site. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis to confirm the loss of the NsiI site and sequenced to confirm the sequence of the (15-125) hIL-3 variant gene. The plasmid, pMON13361, encodes the (15-125) hIL-3 variant with the amino acid sequence of PEPTIDE #25 [SEQ ID NO:89]. DNA sequence # 32 [SEQ ID NO:160] codes for the foregoing pMON13361 polypeptide.

EXAMPLE 58

Construction of pMON14058

Plasmid pMON13361 was mutagenized by oligonucleotide site-directed mutagenesis (method described by Taylor et al., 1985 using a kit from Amersham, Arlington Heights, Ill.) to eliminate a EcoRV site in the (15-125) hIL-3 variant coding region. Codon 46 and 47 of (15-125) hIL-3, encoding asparagine and isoleucine, were converted from GAT to GAC and ATC to ATT respectively, destroying the EcoRV recognition site. The oligonucleotide, Oligo # 55 [SEQ ID NO:159], was used in this procedure to eliminate the EcoRV site. Transformant bacteria were selected on ampicillin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis to confirm the loss of the EcoRV site and sequenced to confirm the sequence of the (15-125) hIL-3 variant gene. The plasmid, pMON14058, encodes the (15-125) hIL-3 variant with the amino acid sequence of PEPTIDE #25 [SEQ ID NO:89]. DNA sequence # 33 [SEQ ID NO:161] codes for the foregoing pMON14058 polypeptide.

EXAMPLE 59

Construction of pMON13438

Plasmid pMON5723 DNA was digested with restriction enzymes NcoI and HindIII resulting in a 3278 NcoI,HindIII fragment. The genetic elements derived from pMON5723 are the pBR327 origin of replication, precA promoter, g10L ribosome binding protein, T7 transcription terminator and streptomycin adenylyltransferase gene. Plasmid pMON14058 DNA was digested with NcoI and HindIII resulting in a 345 base pair NcoI,HindIII fragment which contains the (15-125) hIL-3 gene with the following amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V, 67H, 69E,73G, 76A, 79R, 83Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The restriction fragments were ligated and the ligation reaction mixture was used to transform

E. coli

strain JM101. Transformant bacteria were selected on spectinomycin-containing plates. Plasmid DNA was isolated, analyzed by restriction analysis and sequenced to confirm the correct insert. The plasmid, pMON13438, encodes the (15-125) hIL-3 variant with the amino acid sequence of PEPTIDE #25 [SEQ ID NO:89]. DNA sequence # 33 [SEQ ID NO:161] codes for the foregoing pMON13438 polypeptide.

EXAMPLE 60

Construction of pMON13285

Plasmid pMON13252 DNA was digested with restriction enzymes NcoI and EcoRV and the resulting 3669 base pair NcoI,EcoRV fragment contains the following genetic elements; streptomycin adenyltransferase gene, pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, recA promoter, g10L ribosome binding site and the bases encoding amino acids 47-125 of (15-125) hIL-3 with the following amino acid substitution, 50D. The 3669 base pair NcoI,EcoRV restriction fragment from pMON13252 was ligated to the following annealed complementary oligonucleotides.

Oligo #165 [SEQ ID NO:162]

Oligo #166 [SEQ ID NO:163]

Oligo #167 [SEQ ID NO:164]

Oligo #168 [SEQ ID NO:165]

Oligo #169 [SEQ ID NO:166]

Oligo #170 [SEQ ID NO:167]

When assembled, the oligonucleotides create NcoI and EcoRV restriction ends and the DNA sequence that encodes amino acids 15-46 of (15-125) hIL-3 with the following amino acid substitutions; 42D, 45M and 46S. The codons encoding amino acids 15-46 of (15-125) hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13285, encodes the (15-125) hIL-3 variant with the following amino acid sequence:

Peptide #A3 [SEQ ID NO:259]

DNA sequence #A3 pMON13285 42D, 45M 46S, 50D

[SEQ ID NO:398]

ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC

ACCTGAAGCA GCCACCGCTG CCGCTGCTGG ACTTCAACAA

CCTCAATGAC GAAGACATGT CTATCCTGAT GGACAATAAC

CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA

AGTCTCTGCA GAATGCATCA GCAATTGAGA GCATTCTTAA

AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC

ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG

AATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA

GAACGCGCAG GCTCAACAG

EXAMPLE 61

Construction of pMON13286

Plasmid pMON5978 DNA was digested with restriction enzymes NcoI and EcoRV and the resulting 3865 base pair NcoI,EcoRV fragment contains the following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the bases encoding amino acids 47-125 of (15-125) hIL-3. The 3865 base pair NcoI,EcoRV restriction fragment from pMON5978 was ligated to the following annealed complementary oligonucleotides.

Oligo #165 [SEQ ID NO:162]

Oligo #166 [SEQ ID NO:163]

Oligo #167 [SEQ ID NO:164]

Oligo #168 [SEQ ID NO:165]

Oligo #169 [SEQ ID NO:166]

Oligo #170 [SEQ ID NO:167]

When assembled, the oligonucleotides create NcoI and EcoRV restriction ends and the DNA sequence that encodes amino acids 15-46 of (15-125) hIL-3 with the following amino acid substitutions; 42D, 45M and 46S. The codons encoding amino acids 15-46 of (15-125) hIL-3 are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. The plasmid, pMON13286, encodes the (15-125) hIL-3 variant with the following amino acid sequence:

Peptide #A4 [SEQ ID NO:260]

DNA sequence #A4 pMON13286 42D, 45M, 46S

[SEQ ID NO:399]

ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC

ACCTGAAGCA GCCACCGCTG CCGCTGCTGG ACTTCAACAA

CCTCAATGAC GAAGACATGT CTATCCTGAT GGAAAATAAC

CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA

AGTCTCTGCA GAATGCATCA GCAATTGAGA GCATTCTTAA

AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC

ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG

AATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA

GAACGCGCAG GCTCAACAG

EXAMPLE 62

Construction of pMON13325

The 3704 base pair EcoRI, HindIII DNA fragment from plasmid pMON13286 is ligated to the 64 base pair EcoRI, HindIII DNA fragment from plasmid pMON13215. The following genetic elements are derived from pMON13286; beta-lactamase gene (AMP), pBR327 origin of replication, phage F1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the bases encoding amino acids 15-105 of the (15-125) hIL-3 gene with the following changes, 42D, 45M, and 46S.

The bases encoding amino acids 106-125 of the (15-125) gene with the following change, 116W, are derived from pMON13215. The resulting plasmid, pMON13325, encodes the (15-125) hIL-3 variant with the following amino acid sequence:

Peptide # A5 [SEQ ID NO:261]

EXAMPLE 63

Construction of pMON13326

The 3683 base pair NcoI, EcoRI DNA fragment from plasmid pMON13215 is ligated to the 281 base pair NcoI, EcoRI DNA fragment from plasmid pMON13285. The following genetic elements are derived from pMON13215; beta-lactamase gene (AMP), pBR327 origin of replication, phage F1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the bases encoding amino acids 106-125 of the (15-125) hIL-3 gene with the following change, 116W. The bases encoding amino acids 15-105 of the (15-125) gene with the following change, 42D, 45M, 46S and 50D derived from pMON13285. The resulting plasmid, pMON13326, encodes the (15-125) hIL-3 variant with the following amino acid sequence:

Peptide # A6 [SEQ ID NO:262]

EXAMPLE 64

Construction of pMON13332

Plasmid pMON13326 DNA is digested with restriction enzymes NsiI and EcoRI and the resulting 3853 base pair NsiI,EcoRI fragment contains the following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, recA promoter, g10L ribosome binding site and the bases encoding amino acids 15-71 and 106-125 of (15-125) hIL-3 gene with the following changes 42D, 45M, 46S, 50D and 116W. The 3853 base pair NsiI,EcoRI restriction fragment from pMON13326 is ligated to the following annealed complementary oligonucleotides.

Oligo #15(A) [SEQ ID NO:168]

Oligo #16(A) [SEQ ID NO:169]

In the resulting plasmid the 111 bases between the NsiI and EcoRI restriction sites in the (15-125) hIL-3 gene are replaced with 24 bases from the above mentioned oligonucleotides. This linker also creates a NdeI recognition sequence.

EXAMPLE 65

Construction of pMON13330

The 3846 base pair PstI, EcoRI DNA fragment from plasmid pMON13332 is ligated to the 118 base pair PstI, EcoRI DNA fragment from plasmid pMON13305. The following genetic elements are derived from pMON13332; beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, recA promoter, g10L ribosome binding site and the bases encoding amino acids 15-69 and 106-125 of the (15-125) hIL-3 gene with the following change, 42D, 45M, 46S, 50D and 116W. The bases encoding amino acids 70-105 of the (15-125) gene with the following change, 95R, 98I and 100R are derived from pMON13305. The resulting plasmid, pMON13330, encodes the (15-125) hIL-3 variant with the following amino acid sequence:

Peptide # A7 [SEQ ID NO:263]

EXAMPLE 66

Construction of pMON13329

The 3846 base pair PstI, EcoRI DNA fragment from plasmid pMON13332 is ligated to the 118 base pair PstI, EcoRI DNA fragment from plasmid pMON13304. The following genetic elements are derived from pMON13332; beta-lactamase gene (AMP), pBR327 origin of replication, phage f1 origin of replication as the transcription terminator, recA promoter, g10L ribosome binding site and the bases encoding amino acids 15-69 and 106-125 of the (15-125) hIL-3 gene with the following change, 42D, 45M, 46S, and 116W. The bases encoding amino acids 70-105 of the (15-125) gene with the following change, 98I and 100R are derived from pMON13304. The resulting plasmid, pMON13329, encodes the (15-125) hIL-3 variant with the following amino acid sequence:

Peptide # A8 [SEQ ID NO:406]

EXAMPLE 67

Construction of pMON5853 (

FIG. 6

) which Encodes [Met-(15-133)hIL-3 (Arg

129

)]

Plasmid DNA of pMON5847 (Example 2) was treated with NcoI. The restriction enzyme was inactivated by heat treatment (65° C. for 10 minutes). The DNA was then treated with large fragment of DNA polymerase I (Klenow) in the presence of all four nucleotide precursors. This produces DNA termini with non-overlapping ends. After 5 minutes at 37° C., the polymerase was inactivated by heat treatment at 65° C. for 10 minutes. The DNA was then treated with HpaI, an enzyme which produces non-overlapping termini. The DNA was ethanol precipitated and ligated. The ligation reaction mixture was used to transform competent JM101 cells to ampicillin resistance. Colonies were picked and plasmid DNA was analyzed by restriction analysis. A plasmid designated pMON5853 was identified as one containing a deletion of the amino terminal 14 codons of the hIL-3 gene. The DNA sequence for the junction of the ribosome binding site to the (15-133) hIL-3 gene was determined to be the following:

5′-AAGGAGATATATCCATGAACTGCTCTAAC-3′[SEQ ID NO:400]

M N C S N [SEQ ID NO:401]

The lower line contains the one-letter code for the amino acids specified by the coding sequence of the amino terminus of the 15-133 hIL-3 gene. These are methionine, asparagine, cysteine, serine and asparagine.

When cultures of JM101 cells harboring this plasmid were induced with nalidixic acid, it was found that hIL-3 (15-133) accumulated at levels higher than hIL-3 (pMON5847).

The plasmid, pMON5853, encodes Met-(15-133) hIL-3 (Arg

129

) which has the following amino acid sequence:

[SEQ ID NO:402]

Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile

Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp

Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu

Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala

Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys

Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro

Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg

Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn

Ala Gln Ala Gln Gln Thr Thr Leu Arg Leu Ala Ile

Phe

EXAMPLE 68

Construction of pMON13252

Plasmid, pMON2341, DNA was digested with restriction enzymes NcoI and HindIII resulting in a 3619 base pair NcoI/HindIII fragment. The genetic elements derived from pMON2341 are the beta-lactamase gene (AMP), pBR327 origin of replication F1 phage origin of replication as the transcription terminator, precA, g10L ribosome binding site. The plasmid encoding the hIL-3 (15-125) Asp

(50)

variant, was digested with NcoI and HindIII resulting in a 345 base pair NcoI/HindIII fragment. This 345 Base pair NcoI/HindIII fragment was ligated with the 3619 base pair fragment from pMON2341 and the ligation reaction mixture was used to transform

E. coli

K-12 strain JM101. Plasmid DNA was isolated and screened by restriction analysis using NcoI and HindIII. Positive clones contained a 345 base pair NcoI/HindIII fragment. This construct was designated pMON13252. The plasmid, pMON13252, encodes the (15-125)hIL-3 variant with the following amino acid sequence:

PEPTIDE A10; (15-125)HIL-3 Asp

(50)

pMON13252

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu

[SEQ ID NO:407]

15 20 25

Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly

30 35 40

Glu Asp Gln Asp Ile Leu Met Asp Asn Asn Leu Arg Arg Pro Asn

45 50 55

Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser

60 65 70

Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu

75 80 85

Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly

90 95 100

Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr

105 110 115

Leu Glu Asn Ala Gln Ala Gln Gln

120 125

DNA sequence #A10 pMON13252 50D

[SEQ ID NO:408]

ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC

ACCTGAAGCA GCCACCGCTG CCGCTGCTGG ACTTCAACAA

CCTCAATGGT GAAGACCAAG ATATCCTGAT GGAACAATAAC

CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA

ACTCTCTGCA GAATGCATCA GCAATTGAGA GCATTCTTAA

AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC

ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG

AATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA

GAACGCGCAG GCTCAACAG

EXAMPLES 69-76

The variants in Table 5 were constructed by cassette mutagenesis using methods described in the Materials and Methods and the Examples contained herein, particularly Examples 54-58. Parental plasmid DNA (Table 5), digested with the appropriate restriction enzymes (Table 5), was ligated with the indicated annealed pairs of complementary oligonucleotides (Table 5). The assembled oligonucleotides create appropriate restriction ends and a portion of the (15-125) hIL-3 gene sequence (pMON13288 [SEQ ID NO:100]). Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The oligonucleotides create change(s) in the (15-125) hIL-3 gene which encode the corresponding amino acid substitution(s) in the variant polypeptide (Table 5). The amino acids substitutions in addition to and/or different from those in polypeptide # 25 [SEQ ID NO:89] are indicated in Table 5. The table also shows the plasmid designation (pMON number), DNA sequence identification number for the mutated hIL-3 gene and the identification number for the the resulting variant polypeptide. The biological activity (growth promoting activity in AML 193 cells) for some of the variants in Table 5 is shown in Table 1.

EXAMPLES 77-82

The variants in Table 6 were constructed by methods described in the Materials and Methods and the Examples contained herein, particularly in Examples 60 and 61. Parental plasmid DNA (Table 6), digested with the appropriate restriction enzymes (Table 6), was ligated with the indicated restriction fragment (Table 6). Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The resulting mutated (15-125) hIL-3 genes encode the corresponding amino acid substitutions in the variant polypeptides (Table 6). The amino acids substitutions in addition to and/or different from those in polypeptide # 25 [SEQ ID NO:89] are indicated in Table 6. The table also shows the plasmid designation (pMON number), DNA sequence identification number for the mutated hIL-3 gene and the identification number for the the resulting variant polypeptide. The biological activity (growth promoting activity in AML 193 cells) for some of the variants in Table 6 is shown in Table 1.

EXAMPLE 83

Construction of pMON13368

One of the DNA fragments to construct the plasmid, pMON13368, was generated by site-directed mutagenesis employing PCR techniques described in the Materials and Methods and the Examples contained herein, particularly Example 53. The template for the PCR reaction was plasmid, pMON13289, DNA using the oligonucleotides, Oligo #B13 18I123A25H [SEQ ID NO: 182] and Oligo #B14 2341HIN3 [SEQ ID NO:183], as primers. The resulting DNA product was digested with the restriction enzymes NcoI and HindIII. Upon completion, the digest was heated at 70° C. for 15 minutes to inactivate the enzymes. The restriction fragment was purified by phenol/chloroform extraction and precipitation with equal volume isopropanol in the presence of 2M NH4OAc. The oligonucleotide, Oligo #B13 18I23A25H [SEQ ID NO:182], changes the codon at position 23 of (15-125) hIL-3 variant gene pMON13289 [SEQ ID NO:103] from ‘ATT’ to ‘GCA’ (Ile to Ala). The 3619 base pair NcoI, HindIII restriction fragment from pMON2341 was ligated to the PCR-generated NcoI, HindIII restriction fragment. Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The plasmid, pMON13368, contains the (15-125) hIL-3 variant gene (DNA sequence #B15 [SEQ ID NO:346]) which encodes the (15-125) hIL-3 variant polypeptide with the following amino acid sequence:

Polypeptide #B15 [SEQ ID NO. :278]

EXAMPLE 84

Construction of pMON13380

Plasmid, pMON13368, DNA was digested with restriction enzymes EcoRI and HindIII. The resulting 3900 base pair EcoRI,HindIII fragment contains the following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, phage F1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the DNA sequence encoding amino acids 15-105 of the variant pMON13368. The 3900 base pair EcoRI,HindIII restriction fragment from pMON13368 was ligated to the following annealed complementary oligonucleotides.

Oligo # B48

9E12Q6V1

[SEQ ID NO:217]

Oligo # B49

9E12Q6V3

[SEQ ID NO:218]

Oligo # 49

120Q123E2

[SEQ ID NO:63]

Oligo # 50

120Q123E4

[SEQ ID NO:64]

When assembled, the oligonucleotides create EcoRI and HindIII restriction ends and the DNA sequence that encodes amino acids 106-125 of (15-125) hIL-3 with the following amino acid substitution; 109E, 112Q, 116V, 120Q and 123E. The codons used in the (15-125) hIL-3 gene are those found in the hIL-3 cDNA sequence except at those positions where amino acid substitutions were made. Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The plasmid, pMON13380, contains the (15-125) hIL-3 variant gene (DNA sequence #B16 [SEQ ID NO:347]) which encodes the (15-125) hIL-3 variant polypeptide with the following amino acid sequence:

Polypeptide #B16 [SEQ ID NO.:279]

EXAMPLE 85

Construction of pMON13476

One of the DNA fragments to construct the plasmid, pMON13476, was generated by site-directed mutagenesis employing PCR techniques described in the Materials and Methods and the Examples contained herein, particularly Example 54. The template for the PCR reaction was plasmid, pMON13287, DNA using the oligonucleotides, Oligo #B13 18I23A25H [SEQ ID NO:182] and Oligo #B14 2341HIN3 [SEQ ID NO.:183] as primers. The resulting DNA product was digested with the restriction enzymes NcoI and HindIII. Upon completion, the digest was heated at 70° C. for 15 minutes to inactivate the enzymes. The restriction fragment was purified by phenol/chloroform extraction and precipitation with equal volume isopropanol in the presence of 2M NH4OAc. The oligonucleotide, Oligo #B13 18I23A25H [SEQ ID NO.:182], changes the codon at position 23 of (15-125) hIL-3 variant gene, pMON13287, [SEQ ID NO:97] from ‘ATT’ to ‘GCA’ (Ile to Ala). The 3619 base pair NcoI, HindIII restriction fragment from pMON2341 was ligated to the PCR-generated NcoI, HindIII restriction fragment. Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The resulting clone also contained a change, that was not designed in the mutagenic oligonucleotide, which changed the codon at position −1 from ‘GCT’ to ‘GAT’ which changes the amino acid from Alanine to Aspartic Acid. The plasmid, pMON13476, contains the (15-125) hIL-3 variant gene (DNA sequence #B52 [SEQ ID NO:303]) which encodes the (15-125) hIL-3 variant polypeptide with the following amino acid sequence:

Polypeptide #B52 [SEQ ID NO.:314]

EXAMPLES 86-92

The variants in Table 7 were constructed by PCR techniques using methods described in the Materials and Methods and the Example contained herein, particularly Example 51. Two sequential PCR reactions were used to create the variants. In the first PCR reaction pMON13287 plasmid DNA served as the template and the two oligonucleotides indicated in Table 7 served as the primers. Following the PCR extension reaction, the PCR product was partially purified to remove primer that was not extended. In the second PCR reaction pMON13287 plasmid DNA served as the template, the purified PCR product from the first PCR reaction served as one of the primers and the Oligo #B14 2341Hin3 [SEQ ID NO:183] as the second primer. The product from the second PCR reaction was partially purified and digested with restriction enzymes NcoI and HindIII and ligated with the 3619 base pair NcoI,HindIII fragment from pMON2341. Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The amino acids substitutions in addition to and/or different from those in polypeptide # A 24 [SEQ ID NO:88] are indicated in Table 7. The table also shows the plasmid designation (pMON number), DNA sequence identification number for the mutated hIL-3 gene and the identification number for the the resulting variant polypeptide. The biological activity (growth promoting activity in AML 193 cells) for some of the variants in Table 7 is shown in Table 1.

EXAMPLES 93-120

The variants in Table 8 were constructed by cassette mutagenesis using methods described in the Materials and Methods and the Examples contained here, particularly Examples 54-58. Parental plasmid DNA (Table 8), digested with the appropriate restriction enzymes (Table 8), was ligated with the indicated annealed pairs of complementary oligonucleotides (Table 8). The assembled oligonucleotides create the appropriate restriction ends and a portion of (15-125) hIL-3 gene (pMON13288 [SEQ ID NO:100]) sequence. The oligonucleotides create change(s) in the (15-125) hIL-3 variant gene which encode the corresponding amino acid substitution(s); and/or deletions from the C-terminus of the variant polypeptide (Table 8). Individual isolates were screened by restriction analysis and DNA sequenced to confirm that the desired changes in the (15-125) hIL-3 variant gene were made. The amino acids substitutions in addition to and/or different from those in polypeptide # 25 [SEQ ID NO:88] are indicated in Table 8. The table also shows the plasmid designation (pMON number), DNA sequence identification number for the mutated hIL-3 gene and the identification number for the the resulting variant polypeptide. The biological activity (growth promoting activity in AML 193 cells) for some of the variants in Table 5 is shown in Table 1.

EXAMPLE 121

Construction of pMON13446

Plasmid, pMON13287, DNA (purified from the

E. coli

strain GM48 {dam-}) was digested with restriction enzymes NcoI and ClaI. The resulting 3942 base pair NcoI,ClaI fragment contains the following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, phage F1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the DNA sequence encoding amino acids 21-125 of the (15-125) hIL-3 variant pMON13287. The 3942 base pair NcoI,ClaI restriction fragment from pMON13368 was ligated to the following annealed complementary oligonucleotides.

Oligo #B57

338UP

[SEQ ID NO:226]

Oligo #B56

338D0WN

[SEQ ID NO:225]

When assembled, the oligonucleotides create NcoI and ClaI restriction ends and the DNA sequence that encodes the following 14 amino acid sequence; Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp Asp Lys [SEQ ID NO:403] and the DNA sequence which encodes amino acids 15-20 of the (15-125) hIL-3 variant gene, pMON13287 [SEQ ID NO:97]. The resulting variant polypeptide has a 14 amino acid N-terminal extension fused to the (15-125) hIL-3 variant polypeptide, pMON13288 [SEQ ID NO: 88]. The plasmid, pMON13446, contains the (15-125) hIL-3 variant gene (DNA sequence #B53 [SEQ ID NO:404]) which encodes the (15-125) hIL-3 variant polypeptide with the following amino acid sequence:

Polypeptide #B53 [SEQ ID NO.:315]

EXAMPLE 122

Construction of pMON13390

Plasmid, pMON13288, DNA (purified from the

E. coli

strain GM48 {dam-}) was digested with restriction enzymes NcoI and ClaI. The resulting 3942 base pair NcoI,ClaI fragment contains the following genetic elements; beta-lactamase gene (AMP), pBR327 origin of replication, phage F1 origin of replication as the transcription terminator, precA promoter, g10L ribosome binding site and the DNA sequence encoding amino acids 21-125 of the (15-125) hIL-3 variant pMON13288. The 3942 base pair NcoI,ClaI restriction fragment from pMON13288 was ligated to the following annealed complementary oligonucleotides.

Oligo #B57

338UP

[SEQ ID NO:226]

Oligo #B56

338DOWN

[SEQ ID NO:225]

When assembled, the oligonucleotides create NcoI and ClaI restriction ends and the DNA sequence which encodes the following 14 amino acid sequence; Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp Asp Lys [SEQ ID NO:403] and the DNA sequence which encodes amino acids 15-20 of the (15-125) hIL-3 variant gene pMON13288 [SEQ ID NO:100]. The resulting variant has a 14 amino acid N-terminal extension fused to the (15-125) hIL-3 variant polypeptide, pMON13288 [SEQ ID NO:88]. The plasmid, pMON13390, containes the (15-125) hIL-3 variant gene (DNA sequence #B54 [SEQ ID NO.:405] which encodes the (15-125) hIL-3 variant polypeptide with the following amino acid sequence:

Polypeptide #B54 [SEQ ID NO:316]

EXAMPLES 133-136

The variants in Table 10 were constructed by methods described in Materials and Methods and in Examples contained herein, particularly Examples 54-58. Parental plasmid DNA (Table 10), digested with the appropriate restriction enzymes (Table 10) was ligated with the indicated restriction fragment containing the changes listed (Table 10). The resulting mutated (15-125) IL-3 genes encode the corresponding amino acid substitutions in the variant polypeptides (Table 10). The amino acid substitutions in addition to and/or different from those in polypeptide #25 [SEQ ID NO: 89] are indicated in Table 10. The biological activity (growth promoting activity in AML 193 cells) for some of the variants in Table 10 is shown in Table 1.

EXAMPLES 123-132

The variants in Table 9 were constructed by cassett mutagenesis using methods described in Materials and Methods and in Examples 54-58 contained herein. Parental plasmid DNA (Table 9), digested with the appropriate restriction enzymes (Table 9), was ligated with the indicated annealed pairs of complementry oligonucleoties (Table 9). The assembled oligonucleotides create the appropriate restriction fragment which was inserted into the (15-125) hIL-3 gene (pMON13288 [SEQ ID NO:100] between these restriction sites. The deletions or substitutions encoded by the oligonucleotide in the (15-125) IL-3 gene correspond to the amino acid deletions or substitutions in the variant polypeptide (Table 9). The amino acid substitutions or deletions, in addition to and/or different from those in the polypeptide #25 [SEQ ID NO:89] are indicated in Table 9. The biological activity (growth promoting activity in AML 193 cells) for some of the variants in Table 9 is shown in Table 1.

Formula XI shown below is a representation of a [(15-125)hIL-3 mutein] with numbers in bold type added above the amino acids to represent the position at which the amino acid below the bolded number appears in native (1-133)hIL-3 [e. g. the amino acid at position 1 of Formula XI corresponds to the Asn which appears at position 15 in native (1-133)hIL-3]. The number shown in bold indicates the amino acids that correspond to the native IL-3(1-133). The non-bold members below the amino acids sequences are for Seq Id reference numbers. When the muteins are expressed the initial amino acid may be preceded by Met- or Met-Ala-.

15 20 25

[SEQ ID NO:23]

Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln

1 5 10 15

30 35 40

Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp

20 25 30

45 50 55

Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu

35 40 45

60 65 70

Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile

50 55 60

75 80 85

Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr

65 70 75

90 95 100

Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp

80 85 90

105 110 115

Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu

95 100 105

120 125

Asn Ala Gln Ala Gln Gln

110

TABLE 5

Parental plasmid/

amino acid

resulting

Example

pMON number

restriction digest

oligo pair 1,4

oligo pair 2,5

oligo pair 3,6

changes

polypeptide

Example 69

pMON13406

pMON13288/

19Ala1

29R32N37P2

42S45M3

19Ala

polypeptide B1

SEQ ID NO:332

NcoI, EcoRV

OLIGO#B1

OLIGO#5

OLIGO#11

SEQ ID NO:264

SEQ ID NO:170

SEQ ID NO:19

SEQ ID NO:25

19Ala4

29R32N37P5

42S45M6

OUGO#B2

OLIGO#6

OLIGO#12

SEQ ID NO:171

SEQ ID NO:20

SEQ ID NO:26

Example 70

pMON13414

pMON13288/

19Ile1

29R32N37P2

42S45M3

19Ile

polypeptide B2

SEQ ID NO:333

NcoI, EcoRV

OLIGO#B3

OLIGO#5

OLIGO#11

SEQ ID NO:265

SEQ ID NO:172

SEQ ID NO:19

SEQ ID NO:25

19Ile4

29R32N37P5

42S45M6

OLIGO#B4

OLIGO#6

OLIGO#12

SEQ ID NO:173

SEQ ID NO:20

SEQ ID NO:26

Example 71

pMON13407

pMON13288/

18125H1

29R32N37P2

42S45V3

45Val

polypeptide B3

SEQ ID NO:334

NcoI, EcoRV

OLIGO#1

OLIGO#5

OLIGO#B11

SEQ ID NO:266

SEQ ID NO:15

SEQ ID NO:19

SEQ ID NO:180

18125H4

29R32N37P5

42S45V6

OLIGO#2

OLIGO#6

OLIGO#B12

SEQ ID NO:16

SEQ ID NO:20

SEQ ID NO:181

Example 72

pMON13405

pMON13288/

19Ala1

29R32N37P2

42S45V3

19Ala,45Val

polypeptide B4

SEQ ID NO:335

NcoI, EcoRV

OLIGO#B1

OLIGO#5

OLIGO#B11

SEQ ID NO:267

SEQ ID NO:170

SEQ ID NO:19

SEQ ID NO:180

19Ala4

29R32N37P5

42S45V6

OLIGO#B2

OLIGO#6

OLIGO#B12

SEQ ID NO:171

SEQ ID NO:20

SEQ ID NO:181

Example 73

pMON13415

pMON13288/

19Ile1

29R32N37P2

42S45V3

19Ile,45Val

polypeptide B5

SEQ ID NO:336

NcoI, EcoRV

OLIGO#B3

OLIGO#5

OLIGO#B11

SEQ ID NO:268

SEQ ID NO:172

SEQ ID NO:19

SEQ ID NO:180

19Ile4

29R32N37P5

42S45V6

OLIGO#B4

OLIGO#6

OLIGO#B12

SEQ ID NO:173

SEQ ID NO:20

SEQ ID NO:181

Example 74

pMON13408

pMON13288/

49Ile1

59L62V2

67H69E3

49ILe

polypeptlde B6

SEQ ID NO:337

EcoRV, NsiI

OLIGO#B7

OLIGO#25

OLIGO#29

SEQ ID NO:269

SEQ ID NO:176

SEQ ID NO:39

SEQ ID NO:43

49Ile4

59L62V5

67H69E6

OLIGO#B8

OLIGO#26

OLIGO#30

SEQ ID NO:177

SEq ID NO:40

SEQ ID NO:44

n

Example 75

pMON13409

pMON13288/

49Leu1

59L62V2

67H69E3

49Leu

polypeptide B7

SEQ ID NO:338

EcoRV, NsiI

SEQ ID NO:178

OLIGO#25

OLIGO#29

SEQ ID NO:270

OLIGO#B9

SEQ ID NO:39

SEQ ID NO:43

49Leu4

59L612V5

67H69E6

OLIGO#B10

OLIGO#26

OLIGO#30

SEQ ID NO:179

SEQ ID NO:40

SEQ ID NO:44

Example 76

PMON1341O

pMON13288/

49Asp1

59L62V2

67H69E3

49Asp

polypeptide

SEQ ID NO:339

EeoRV, NsiI

OLIGO#B5

OLIGO#25

OLIGO#29

SEQ ID NO:271

SEQ ID NO:174

SEq ID NO:39

SEQ ID NO:43

49Asp4

59L62V5

67H69E6

OLIGO#B6

OLIGO#26

OLIGO#30

SEQ ID NO:175

SEQ ID NO:40

SEQ ID NO:44

TABLE 6

plasmid pMON

Parental plasmid/

restriction

amino add

resulting

Example No

number

restriction digest

fragment

substitutions

polypeptide

Example 77

pMON13422

pMON13408/

99 base pair

19Ala,

polypeptide B9

SEQ ID NO:340

NcoI, EcoRV

NcoI, EcoRV

45Val,

SEQ ID NO:272

fragment from

49Ile

pMON13405

Example 78

pMON13423

pMON13408/

99 base pair

19Ile,

polypeptide B10

SEQ ID NO:341

NcoI, EcoRV

NcoI, EcoRV

45Val,

SEQ ID NO:273

fragrament from

49Ile

pMON13415

Example 79

pMON13424

PMON13409/

99 base pair

19Ala,

polypeptide B11

SEQ ID NO:342

NcoI, EcoRV

NcoI, EcoRV

45Val,

SEQ ID NO:274

fragment from

49Leu

pMON13405

Example 80

pMON13425

pMON13409/

99 base pair

19Ile,

polypeptide B12

SEQ ID NO:343

NcoI, EcoRV

NcoI, EcoRV

45Val,

SEQ ID NO:275

fragment from

49Leu

pMON13415

Eumple 81

pMON13426

pMON13410/

99 base pair

19Ala,

polypeptide B13

SEQ ID NO:344

NcoI, EcoRV

NcoI, EcoRV

45Val,

SEQ ID NO:276

fragment from

49Asp

pMON13405

Example 82

pMON13429

pMON13410/

99 base pair

19Ile,

polypeptide B14

SEQ ID NO:345

NcoI, EcoRV

NcoI, EcoRV

45Val,

SEQ ID NO:277

fragment from

49Asp

pMON13415

TABLE 7

pMON

Step one

Step one

Step two

Step two

Amino Acid

Example

number

template

PCR primer1

PCR primer2

PCR primer1

PCR primer2

Substitutions

Polypeptide

Example 86

pMON13475

pMON13287

18I23A25H

42D45V46S50D

product from

2341HIN3

23A,46S,42D,

Polypeptide

SEQ ID NO:

OLIGO#B13

OLIGO#B19

step one

OLIGO#B14

50D

#B17

348

SEQ ID NO:182

SEQ ID NO:188

SEQ ID NO:183

SEQ ID NO 280

Example 87

pMON13366

pMON13287

2341NCO

42D45V46S50D

product from

2341HIN3

42N,46S,50D

Polypeptide

SEQ ID NO:

OLIGO#B15

OLIGO#B19

step one

OLIGO#B14

#B18

349

SEQ ID NO:184

SEQ ID NO:188

SEQ ID NO:183

SEQ ID NO 281

Example 88

pMON13367

pMON13287

2341NCO

42A45V46S50D

product from

2341HIN3

46S,50D

Polypeptide

SEQ ID NO:

OLIGO#B15

OLIGO#B17

step one

OLIGO#B14

#B19

350

SEQ ID NO:184

SEQ ID NO:186

SEQ ID NO:183

SEQ ID NO 282

Example 89

pMON13369

pMON13287

2341NCO

42D45V46S50D

product from

2341HIN3

42D,46S,50D

Polypeptide

SEQ ID NO:

OLIGO#B15

OLIGO#B21

step one

OLIGO#B14

#B20

351

SEQ ID NO:184

SEQ ID NO:190

SEQ ID NO:183

SEQ ID NO 283

Example 90

pMON13370

PMON13287

2341NCO

42A45M46S50D

product from

2341HIN3

45M,46S,50D

Polypeptide

SEQ ID NO:

OLIGO#B15

OLIGO#B16

step one

OLIGO#B14

#B21

352

SEQ ID NO:184

SEQ ID NO:185

SEQ ID NO:183

SEQ ID NO 284

Example 91

pMON13373

pMON13287

2341NCO

42D45M46S50D

product from

2341HIN3

42D,45M,46S

Polypeptide

SEQ ID NO:

OLIGO#B15

OLIGO#B18

step one

OLIGO#B14

50D

#B22

353

SEQ ID NO:184

SEQ ID NO:187

SEQ ID NO:183

SEQ ID NO 285

Example 92

pMON13374

pMON13287

2341NCO

42S45M46S50D

product from

2341HIN3

42S,45M,46S

Polypeptide

SEQ ID NO:

OLIGO#B15

OLIGO#B20

step one

OLIGO#B14

50D

#B23

354

SEQ ID NO:184

SEQ ID NO:189

SEQ ID NO:183

SEQ ID NO 286

TABLE 8

parental

resulting amino

Example

plasmid

plasmid

oligo pair

oligo pair

oligo pair

oligo pair

acid sub(s)

polypeptide

Example

pMON13375

pMON13287/

S09E16V1

S116VD31

15-119

polypeptide

93

SEQ ID NO:

EcoR1,HindIII

OLIGO#B50

OLIGO#B52

B24 SEQ ID

355

SEQ ID NO:219

SEQ ID NO:221

NO:397

S09E16V3

SECR1D33

OLIGO#B51

OLIGO#B53

SEQ ID NO:220

SEQ ID NO:222

Example

pMON13376

pMON13476/

S9E2Q6V1

S116VD31

15-119,23A,

polypeptide

94

SEQ ID NO:

EcoR1,HindIII

OLIGO#B54

OLIGO#B52

112Q

B25 SEQ ID

356

SEQ ID NO:223

SEQ ID NO:221

NO: 288

59E2Q6V3

SECR1D33

OLIGO#B55

OLIGO#B53

SEQ ID NO:224

SEQ ID NO:222

Example

pMON13377

pMON13475/

S9E2Q6V1

S116VD31

15-119,23A,

polypeptide

95

SEQ ID NO:

EcoR1,HindIII

OLIGO#B54

OLIGO#B52

42D,46S,50D,

B26 SEQ ID

357

SEQ ID NO:223

SEQ ID NO:221

112Q

NO:289

59E2Q6V3

SECRID33

OLIGO#B55

OLIGO#B53

SEQ ID NO:224

SEQ ID NO:222

Example

pMON13378

pMON13365/

S09E16V1

S116VD31

15-119,23A

polypeptide

96

SEQ ID NO:

EcoR1,HindIII

OLIGO#B50

OLIGO#B52

B27 SEQ ID

358

SEQ ID NO:219

SEQ ID NO:221

NO:290

SQ9E16V3

SECR1D33

OLIGO#B51

OLIGO#B53

SEQ ID NO:220

SEQ ID NO:222

Example

pMON13379

pMON13367/

9E12Q6V1

120Q123E2

46S,50D,112Q

polypeptide

97

SEQ ID NO:

EcoR1,HindIII

OLIGO#B48

OLIGO#49

B28 SEQ ID

359

SEQ ID NO:217

SED ID NO:63

NO:291

9E12Q6V3

12QQ123E4

OLIGO#B49

OLIGO#50

SEQ ID NO:218

SED ID NO:64

Example

pMON13385

pMON13287/

18I25H1

29V32R34S2

42A45V3

29V,32R,34S

polypeptide

98

SEQ ID NO:

NcoI,EcoRV

OLIGO#1

OLIGO#B28

OLIGO#9

B29 SEQ ID

360

SEQ ID NO:15

SEQ ID NO:197

SEQ ID NO:23

NO:292

18I25H4

29V32R34D5

42A45V6

OLIGO#2

OLIGO#B29

OLIGO#10

SEQ ID NO:16

SED ID NO:198

SEQ ID NO:24

Example

pMON13381

pMON132B7/

73G76A1

82TRP2

87S93S98I3

101A105Q4

82W

polypeptide

99

SEQ ID NO:

NsiI,EcoRI

OLIGO#41

OLIGO#B44

OLIGO#35

OLIGO#43

B30 SEQ ID

361

SEQ ID NO:55

SEQ ID NO:213

SEQ ID NO:49

SEQ ID NO:57

NO:293

73G76A4

82TRP5

87S93S98I7

101A105Q88

OLIGO#42

OLIGO#B45

OLIGO#36

OLIGO#44

SEQ ID NO:56

SEQ ID NO:214

SEQ ID NO:50

SEQ ID NO:58

Example

pMON13383

pMON13475/

9E12O6V1

120Q123E2

23A,42D,46S,

polypeptide

100

SEQ ID NO:

EcoR1,NindIII

OLIGO#B48

OLIGO#49

50D,112Q

B31 SEQ ID

362

SEQ ID NO:217

SED ID NO:63

NO:294

9E12Q6V5

12QQ123E4

OLIGO#B49

OLIGO#50

SEQ ID NO:218

SED ID NO:64

Example

pMON13384

pMON13287/

9E12Q6V1

120Q123E2

112Q

polypeptide

101

SEQ ID NO:

EcoR1,HindIII

OLIGO#B48

OLIGO#49

B32 SEQ ID

363

SEQ ID NO:217

SED ID NO:63

NO:295

9E12QEV5

120Q123E4

OLIGO#B49

OLIGO#50

SEQ ID NO:218

SED ID NO:64

Example

pMON13388

pMON13287/

50D56S1

60S62V2

67N69E3

50D,56S

polypeptide

102

SEQ ID NO:

EcoRV,NsiI

OLIGO#B42

OLIGO#27

OLIGO#31

B33 SEQ ID

364

SEQ ID NO:211

SEQ ID NO:41

SEQ ID NO:45

NO:296

50ASP4

56SERS

67N69E6

OLIGO#B41

OLIGO#B43

OLIGO#32

SEQ ID NO:210

SEQ ID NO:212

SEQ ID NO:46

Example

pMON13389

pMON13287/

18I25H1

29R32A37P2

42D45N3

42D,45M

polypeptide

103

SEQ ID NO:

NcoI,ECORV

OLIGO#1

OLIGO#3

OLIGO#B32

B34 SEQ ID

365

SEQ ID NO:15

SEQ ID NO:17

SEQ ID NO:201

NO:297

18I25H4

29R32A37P5

42D45N6

OLIGO#2

OLIGO#4

OLIGO#B33

SEQ ID NO:16

SEQ ID NO:18

SEQ ID NO:202

Example

pMON13391

pMON13287/

18I25H1

34SER1

42A45V3

34S

polypeptide

104

SEQ ID NO:

NcoI,EcORV

OLIGO#1

OLIGO#B30

OLIGO#9

B35 SEQ ID

366

SEQ ID NO:15

SEQ ID NO:199

SEQ ID NO:23

NO:298

18I25H4

34SERS

42A45V6

OLIGO#2

OLIGO#B31

OLIGO#10

SEQ ID NO:16

SEQ ID NO:200

SEQ ID NO:24

Example

pMON13392

pMON13287/

18I25H1

29R32A37P2

42D45V3

42D

polypeptide

105

SEQ ID NO:

NcoI,EcoRV

OLIGO#1

OLIGO#3

OLIGO#B34

B36 SEQ ID

367

SEQ ID NO:15

SEQ ID NO:17

SEQ ID NO:203

NO:299

18I25H4

29R32A37P5

42D45V6

OLIGO#2

OLIGO#4

OLIGO#B35

SEQ ID NO:16

SEQ ID NO:18

SEQ ID NO:204

Example

pMON13393

pMON13287/

23ALA1

34SER1

42D45M46S3

23A,34S,42D

polypeptide

106

SEQ ID NO:

NcoI,EcoRV

OLIGO#B26

OLIGO#30

OLIGO#B36

45M,46S

B37 SEQ ID

368

SEQ ID NO:195

SEQ ID NO:199

SEQ ID NO:205

NO:300

23ALA4

34SER5

42D45M4656

OLIGO#B27

OLIGO#B31

OLIGO#B37

SEQ ID NO:196

SEQ ID NO:206

SEQ ID NO:206

Example

pMON13394

pMON13287/

18I25H1

29R32A37P2

42D45M4653

42D,45M,45S

polypetide

107

SEQ ID NO:

NcoI, EcoRV

OILGO#1

OLIGO#3

OLIGO#B36

B38 SEQ ID

369

SEQ ID NO:15

SEQ ID NO:17

SEQ ID NO:205

NO:301

18125N4

29R32A37P5

42D45N4656

OLIGO#2

OLIGO#4

OLIGO#B37

SEQ ID NO:16

SEQ ID NO:18

SEQ ID ND:206

Example

pMON13395

pMON13287/

23ALA1

29V32R34S2

42D45V46S3

23A,29V,32R,

polypeptide

108

SEQ ID NO:

NcoI,EcoRV

OLIGO#B26

OLIGO#B28

OLIGo#B38

34S,42D,46S

B39 SEQ ID

370

SEQ ID NO:195

SED ID NO:197

SEQ ID NO:207

NO:302

23ALA4

29V32R34S5

42D45V46S6

OLIGO#B27

OLIGO#B29

OLIGO#B39

SEQ ID NO:196

SED ID NO:198

SEQ ID NO:208

Example

pMON13396

pMON132871

73G76A1

79R82Q2

100ARG3

100NET4

IGCR.101N

polypeptide

109

SEQ ID NO:

NcoI,EcoRV

OLIGO#41

OLIGO#B39

SEQ ID NO:

OLIGO#B24

B40 SEQ ID

371

SEQ ID NO:55

SEQ ID NO:53

87S93S98I7

SEQ ID NO:193

NO:303

73G76A4

79R82Q5

OLIGO#36

10R01M8

OLIGO#42

OLIGO#B40

SEQ ID NO:50

OLIGO#B25

SEQ ID NO:56

SEQ ID NO:54

SEQ ID NO:194

Example

pMON13397

pMON132871

73C76A1

82TRP2

100ARC3

100MET4

82M,100R,

polypeptide

110

SEQ ID NO:

Ncol,EcoRV

OLIGO#41

OLIGO#B44

OLIGO#B22

OLIGO#B24

101M

B41 SEQ ID

372

SEQ ID NO:55

SEQ ID NO:213

SEQ ID NO:191

SEQ ID NO:193

NO:304

73G7EA4

82TRPS

87S93S98I7

10R01M8

OLIGO#42

OLIGO#B45

OLIGO#36

OLIGO#B25

SEQ ID NO:56

SEQ ID NO:214

SEQ ID NO:50

SEQ ID NO:194

Example

pMON13398

pMON13287/

18I25H1

29R32A37P2

42D45V46S3

42D,465

polypeptide

111

SEQ ID NO:

NcoI,EcoRV

OLIGO#81

OLIGO#3

OLIGO#B38

B42 SEQ ID

373

SEQ ID NO:15

SEQ ID NO:17

SEQ ID NO:207

NO:305

18I25H4

29P32A37P5

42D45V46S6

OLIGO#2

OLIGO#4

OLIGO#B39

SEQ ID NO:16

SEQ ID NO:18

SEQ ID NO:208

Example

pMON13399

pMON13388/

23ALA1

29V32R34S2

42D45V46S3

23A,29V,32R,

polypeptide

112

SEQ ID NO:

NcoI,EcoRV

OLIGO#B26

OLIGO#B28

OLIGO#B38

34D,42D,46S

B43 SEQ ID

374

SEQ ID NO:195

SED ID NO:197

SEQ ID NO:207

NO:306

23ALA4

29V32R34S5

42D45V46S6

OLIGO#B27

OLIGO#B29

OLIGO#B39

SEQ ID NO:196

SED ID NO:198

SEQ ID NO:208

Example

pMON13404

pMON13287/

59E2Q6V1

S116VD31

15-119

polypeptide

113

SEQ ID NO:

EcoRI,HindIII

OLIGO#B54

OLIGO#B52

1120

B44 SEQ ID

375

SEQ ID NO:223

SEQ ID NO:221

NO:307

S9E2Q6V3

SECRID33

OLIGO#B55

OLIGO#53

SEQ ID NO:221

SEQ ID NO:222

Example

pMON13387

pMON3287/

SOASP1

6QS62V2

67N69E3

50D

polypeptide

114

SEQ ID NO:

EcORV,NsiI

OLIGO#940

OLIGO#27

OLIGO#31

B45 SEQ ID

376

SEQ ID NO:209

SEQ ID NO:41

SEQ ID NO:45

NO:308

S0ASP4

60S62V5

67N69E6

OLIGO#B41

OLIGO#28

OLIGO#32

SEQ ID NO:210

SEQ ID NO:42

SEQ ID NO:46

Example

pMON13416

pMON13387/

18I25H1

29R32A37P2

42D41V46S3

42D,46S,50D

polypeptide

115

SEQ ID NO:

NcoI,EcoRV

OLIGO#1

OLIGO#3

OLIGO#B38

B46 SEQ ID

377

SEQ ID NO:15

SEQ ID NO:17

SEQ ID NO:207

NO:309

18t25N4

29R32A37P5

42D45V4656

OLIGO#2

OLIGO#4

OLIGO#B35

SEQ ID NO:16

SEQ ID NO:18

SEQ ID NO:208

Example

pMON13417

pMON13387/

18I2SH1

29R32A37P2

42D45M46S3

42D,45N,46S,

polypeptide

116

SEQ ID NO:

NcoI,EcoRV

OLIGO#1

OLIGO#3

OLIGO#B36

50D

B47 SEQ ID

378

SEQ ID NO:15

SEQ ID NO:17

SEQ ID NO:205

NO:310

18I25H4

29R32A37P5

42D45M46S6

OLIGO#2

OLIGO#4

OLIGO#B37

SEQ ID NO:16

SEQ ID NO:18

SEQ ID NO:206

Example

pMON13420

pMON13388/

23ALA1

345ER1

42D45V4653

23A,34S,42D,

polypeptide

117

SEQ ID NO:

NcoI,EcoRV

OLIGO#B26

OLIGO#B30

OLIGO#B38

46S,50D,565

B48 SEQ ID

379

SEQ ID NO:195

SEQ ID NO:199

SEQ ID NO:207

NO:311

23ALA4

34SER5

42D45V46S6

OLIGO#B27

OLIGO#B31

OLIGO#B39

SEQ ID NO:196

SEQ ID NO:200

SEQ ID NO:208

Example

pMON13421

pMON13388/

23ALA1

34SER1

42D41M46S3

23A,34S,42D,

polypeptide

118

SEQ ID NO:

NcoI,EcoRV

OLIGO#B26

OLIGO#B30

OLIGO#B36

45N,46S,

B49 SEQ ID

380

SEQ ID NO:195

SEQ ID NO:199

SEQ ID NO:205

50D,56S

NO:331

23ALA4

34SER5

42D45M46S6

OLIGO#B27

OLIGO#B31

OLIGO#B37

SEQ ID NO:196

SEQ ID NO:200

SEQ ID NO:206

Example

pMON13432

pMON133B7/

23ALAT

345ER1

42D45N4653

23A,34S,42D,

polypeptide

119

SEQ ID NO:

NcoI,EcoRV

OILGO#B26

OLIGO#B30

OLIGO#B36

45M,465,50D

B50 SEQ ID

381

SEQ ID NO:195

SEQ ID NO:199

SEQ ID NO:205

NO:312

23ALA4

34SER5

42D45M46S6

OLIGO#B27

OLIGO#B31

OLIGO#B37

SEQ ID NO:196

SEQ ID NO:200

SEQ ID NO:206

Example

pMON13382

pMON13287/

9E12Q6W1

120Q123E2

112Q,11EW

polypeptide

120

SEQ ID NO:

EcoRI,HindIII

OLIGO#B46

OLIGO#49

B51 SEQ ID

382

SEQ ID NO:215

SED ID NO:63

NO:313

9E12Q16W3

120Q123E4

OLIGO#41

OLIGO#50

SEQ ID NO:216

SEQ ID NO:64

TABLE 9

Parental

Plasmid/

Example

Restriction

Amino acid

No.

Plasmid

Digest

Oligo pair

Oligo pair

Oligo pair

Oligo pair

changes

Polypeptide

Example

pMON13400

pMON13288

20P23Al

29I4S752

38A5V6S3

20P 23A

Polypeptide

124

SEQ ID NO:

Restriction

SEQ ID NO:232

SEQ ID NO:236

SEQ ID NO:238

29I 34S

C-2 SEQ ID

384

NcoI-EcoRV

20P23A4

29I4S7S5

38A5V6S3

37S 38A

NO:317

SEQ ID NO:233

SEQ ID NO:237

SEQ ID NO:239

45V 46S

Example

pMON13402

pMON13288

23L1

29I4S7S2

38A5V6S3

23L 29I

Polypeptide

125

SEQ ID NO:

Restriction

SEQ ID NO:234

SEQ ID NO:236

SEQ ID NO:238

34S 37S

C-3 SEQ ID

385

NcoI-EcoRV

23IA

29I4S7S5

38A5V6S3

38A 45V

NO:318

SEQ ID NO:235

SEQ ID NO:237

SEQ ID NO:239

46S

Example

pMON13440

pMON13288

18I3A5H1

29I4S752

38A5V6S3

18I 23A 25H

Polypeptide

131

SEQ ID NO:

Restriction

SEQ ID NO:195

SEQ ID NO:236

SEQ ID NO:238

29I 34S 37S

C-10 SEQ ID

386

NcoI-EcoRV

18I3A5H4

29I4S7S5

38A5V6S3

38A 45V

NO:319

SEQ ID NO:196

SEQ ID NO:237

SEQ ID NO:239

46S

Example

pMON13451

pMON13288

19IOL3A1

29I4S7S2

38A5V6S3

19I 20L 23A

Polypeptide

132

SEQ ID NO:

Restriction

SEQ ID NO:230

SEQ ID NO:236

SEQ ID NO:238

29I 34S 37S

C-11 SEQ ID

387

NcoI-EcoRV

19I0L3A4

29I4S7S5

38A5V6S3

38A 45V

NO:320

SEQ ID NO:231

SEQ ID NO:237

SEQ ID NO:239

46S

Example

pMON13419

pMON13288

50D5151

62P3H5S2

67Q3

50D 51S 62P

Polypeptide

130

SEQ ID NO:

Restriction

SEQ ID NO:240

SEQ ID NO:244

SEQ ID NO:248

63H

C-8 SEQ ID

389

EcoRV-NsiI

50D51S4

62P3H5

65S67Q6

65S 67Q

NO:325

SEQ ID NO:241

SEQ ID NO:246

SEQ ID NO:247

Example

pMON13403

pMON13288

50D51S1

62P3H2

67Q3

50D 51S

Polypeptide

126

SEQ ID NO:

Restriction

SEQ ID NO:240

SEQ ID NO:245

SEQ ID NO:248

62P 63H

C-4 SEQ ID

388

EcoRV-NsiI

50D51S4

62P3H5

67Q6

67Q

NO:321

SEQ ID NO:241

SEQ ID NO:246

SEQ ID NO:249

Example

pMON13418

pMON13288

76P1

5VYWPTT3

10YM05Qt

76P 79B

Polypeptide

123

SEQ ID NO:

Restriction

SEQ ID NO:250

SEQ ID NO:252

SEQ ID NO:242

SEQ ID NO:57

8SV 8TY

C-1 SEQ ID

393

NsiI-EcoRI

76P5

79S6

5VYWPTT7

101A10508

88W91P

NO:326

SEQ ID NO:251

SEQ ID NO:253

SEQ ID NO:243

SEQ ID NO:58

95T 98T

Example

pMON13411

pMON13288

09L2Q6S1

120Q123E2

109L 112Q

Polypeptide

127

SEQ ID NO:

Restriction

Seq ID NO:227

SEQ ID NO:63

116S

C-5 SEQ ID

390

EcoRI-HindIII

09L2Q6S3

120Q123E4

NO:322

SEQ ID NO:228

SEQ ID NO:64

Example

pMON13412

pMON13288

9LQS1181

15-118

Polypeptide

128

SEQ ID NO:

Restriction

Seq ID NO:255

109L 112Q

C-6 SEQ ID

391

EcoRI-HindIII

9LQS1183

116S

NO:323

SEQ ID NO:256

Example

pMON13413

pMON13288

09L2Q6S1

11752

109L 112Q

Polypeptide

129

SEQ ID NO:

Restriction

Seq ID NO:227

SEQ ID NO:229

118S 117S

C-7 SEQ ID

392

EcoRI-HindIII

09L2Q6S3

120Q123E4

NO:324

SEQ ID NO:228

SEQ ID NO:64

TABLE 10

Parental

plasmid/

Example

Restriction

Restriction

Amino Acid

No

Plasmid

digest

fragment

changes

Polypeptide

Example 133

pMON13428

pMON13411

102 bp

76P 79S 85V

Polypeptide

SEQ ID

Nsil-EcoRI

Nsil-EcoRI

87Y 91P 95T

C-9

NO:394

fragment from

98T 109L

SEQ ID

pMON13418

112Q 116S

NO:327

Example 134

pMON13459

pMON13428

170 bp

23L 29I 34S

Polypeptide

SEQ ID

NcoI-Nsil

NcoI-Nsil

37S 38A 45V

C-12

NO:395

fragment from

46S 76P 79S

SEQ ID

pMON13402

85V 87Y 91P

NO:328

95T 98T 109L

112Q 116S

Example 135

pMON13467

pMON13413

170 bp

23L 291 34S

Polypeptide

SEQ ID

NcoI-Nsil

NcoI-Nsil

37S 38A 45V

C-13

NO:396

fragment from

46S 109L

SEQ ID

pMON13402

112Q 116S

NO:329

109L 112Q

116S 117S

Example 136

pMON13492

pMON13418

170 bp

23L 29I 34S

Polypeptide

SEQ ID

NcoI-Nsil

NcoI-Nsil

37S 38A 45V

C-14

NO:397

fragment from

46S 76P 79S

SEQ ID

pMON13402

85V 87Y 91P

NO:330

95T 98T

415

23 base pairs

nucleic acid

single

linear

DNA (synthetic)

1
CTAGCGATCT TTTAATAAGC TTG 23

23 base pairs

nucleic acid

single

linear

DNA (synthetic)

2
GATCCAAGCT TATTAAAAGA TCG 23

69 base pairs

nucleic acid

single

linear

DNA (synthetic)

3
GGCAACAATT TCTACAAAAC ACTTGATACT GTATGAGCAT ACAGTATAAT TGCTTCAACA 60
GAACAGATC 69

67 base pairs

nucleic acid

single

linear

DNA (synthetic)

4
TGTTCTGTTG AAGCAATTAT ACTGTATGCT CATACAGTAT CAAGTGTTTT GTAGAAATTG 60
TTGCCGC 67

23 base pairs

nucleic acid

single

linear

DNA (synthetic)

5
CCATTGCTGC CGGCATCGTG GTC 23

46 base pairs

nucleic acid

single

linear

DNA (synthetic)

6
CATGGCTCCA ATGACTCAGA CTACTTCTCT TAAGACTTCT TGGGTT 46

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

7
AACCCAAGAA GTCTTAAGAG AAGTAGTCTG AGTCATTGGA GC 42

64 base pairs

nucleic acid

single

linear

DNA (synthetic)

8
AATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAGT 60
AATA 64

64 base pairs

nucleic acid

single

linear

DNA (synthetic)

9
AGCTTATTAC TGTTGAGCCT GCGCGTTCTC CAAGGTTTTC AGATAGAAGG TCAGTTTACG 60
ACGG 64

126 amino acids

amino acid

linear

peptide

10
Met Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn
1 5 10 15
Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro Pro
20 25 30
Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile
35 40 45
Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg
50 55 60
Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys
65 70 75 80
Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His
85 90 95
Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu
100 105 110
Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
115 120 125

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

11
CATGGCTAAC TGCTCTAACA TGAT 24

22 base pairs

nucleic acid

single

linear

DNA (synthetic)

12
CGATCATGTT AGAGCAGTTA GC 22

113 amino acids

amino acid

linear

peptide

13
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp
20 25 30
Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

27 amino acids

amino acid

linear

peptide

14
Met Met Ile Thr Leu Arg Lys Leu Pro Leu Ala Val Ala Val Ala Ala
1 5 10 15
Gly Val Met Ser Ala Gln Ala Met Ala Asn Cys
20 25

133 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- may or may not precede the
amino acid in position 1”

Modified-site

/note= “Xaa at position 17 is Ser,
Lys, Gly, Asp, Met, Gln, or Arg”

Modified-site

/note= “Xaa at position 18 is Asn,
His, Leu, Ile, Phe, Arg, or Gln”

Modified-site

/note= “Xaa at positiion 19 is Met,
Phe, Ile, Arg, Gly, Ala, or Cys”

Modified-site

/note= “Xaa at position 20 is Ile,
Cys, Gln, Glu, Arg, Pro, or Ala”

Modified-site

/note= “Xaa at position 21 is Asp,
Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn, Thr, Ser,
or Val”

Modified-site

/note= “Xaa at position 22 is Glu,
Trp, Pro, Ser, Ala, His, Asp, Asn, Gln, Leu, Val,
or Gly”

Modified-site

/note= “Xaa at position 23 is Ile,
Val, Ala, Leu, Gly, Trp, Lys, Phe, Ser, or
Arg”

Modified-site

/note= “Xaa at position 24 is Ile,
Gly, Val, Arg, Ser, Phe, or Leu”

Modified-site

/note= “Xaa at position 25 is Thr,
His, Gly, Gln, Arg, Pro, or Ala”

Modified-site

/note= “Xaa at position 26 is His,
Thr, Phe, Gly, Arg, Ala, or Trp”

Modified-site

/note= “Xaa at position 27 is Leu,
Gly, Arg, Thr, Ser, or Ala”

Modified-site

/note= “Xaa at position 28 is Lys,
Arg, Leu, Gln, Gly, Pro, Val, or Trp”

Modified-site

/note= “Xaa at position 29 is Gln,
Asn, Leu, Pro, Arg, or Val”

Modified-site

/note= “Xaa at position 30 is Pro,
His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys”

Modified-site

/note= “Xaa at position 31 is Pro,
Asp, Gly, Ala, Arg, Leu, or Gln”

Modified-site

/note= “Xaa at position 32 is Leu,
Val, Arg, Gln, Asn, Gly, Ala, or Glu”

Modified-site

/note= “Xaa at position 33 is Pro,
Leu, Gln, Ala, Thr, or Glu”

Modified-site

/note= “Xaa at position 34 is Leu,
Val, Gly, Ser, Lys, Glu, Gln, Thr, Arg, Ala, Phe,
Ile, or Met”

Modified-site

/note= “Xaa at position 35 is Leu,
Ala, Gly, Asn, Pro, Gln, or Val”

Modified-site

/note= “Xaa at position 36 is Asp,
Leu, or Val”

Modified-site

/note= “Xaa at position 37 is Phe,
Ser, Pro, Trp, or Ile”

Modified-site

/note= “Xaa at position 38 is Asn,
or Ala”

Modified-site

/note= “Xaa at position 40 is Leu,
Trp, or Arg”

Modified-site

/note= “Xaa at position 41 is Asn,
Cys, Arg, Leu, His, Met, or Pro”

Modified-site

/note= “Xaa at position 42 is Gly,
Asp, Ser, Cys, Asn, Lys, Thr, Leu, Val, Glu, Phe, Tyr,
Ile, Met, or Ala”

Modified-site

/note= “Xaa at position 43 is Glu,
Asn, Tyr, Leu, Phe, Asp, Ala, Cys, Gln, Arg, Thr, Gly,
or Ser”

Modified-site

/note= “Xaa at position 44 is Asp,
Ser, Leu, Arg, Lys, Thr, Met, Trp, Glu, Asn, Gln, Ala,
or Pro”

Modified-site

/note= “Xaa at position 45 is Gln,
Pro, Phe, Val, Met, Leu, Thr, Lys, Trp, Asp, Asn, Arg,
Ser, Ala, Ile, Glu, or His”

Modified-site

/note= “Xaa at position 46 is Asp,
Phe, Ser, Thr, Cys, Glu, Asn, Gln, Lys, His, Ala, Tyr,
Ile, Val, or Gly”

Modified-site

/note= “Xaa at position 47 is Ile,
Gly, Val, Ser, Arg, Pro, or His”

Modified-site

/note= “Xaa at position 48 is Leu,
Ser, Cys, Arg, Ile, His, Phe, Glu, Lys, Thr, Ala, Met,
Val, or Asn”

Modified-site

/note= “Xaa at position 49 is Met,
Arg, Ala, Gly, Pro, Asn, His, or Asp”

Modified-site

/note= “Xaa at position 50 is Glu,
Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala, Ile, Val, His,
Phe, Met, or Gln”

Modified-site

/note= “Xaa at position 51 is Asn,
Arg, Met, Pro, Ser, Thr, or His”

Modified-site

/note= “Xaa at position 52 is Asn,
His, Arg, Leu, Gly, Ser, or Thr”

Modified-site

/note= “Xaa at position 53 is
Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, or Met”

Modified-site

/note= “Xaa at position 54 is Arg,
Asp, Ile, Ser, Val, Thr, Gln, Asn, Lys, His, Ala,
or Leu”

Modified-site

/note= “Xaa at position 55 is Arg,
Thr, Val, Ser, Leu, or Gly”

Modified-site

/note= “Xaa at position 56 is Pro,
Gly, Cys, Ser, Gln, Glu, Arg, His, Thr, Ala, Tyr,
Phe, Leu, Val, or Lys”

Modified-site

/note= “Xaa at position 57 is Asn
or Gly”

Modified-site

/note= “Xaa at position 58 is Leu,
Ser, Asp, Arg, Gln, Val, or Cys”

Modified-site

/note= “Xaa at position 59 is Glu,
Tyr, His, Leu, Pro, or Arg”

Modified-site

/note= “Xaa at position 60 is Ala,
Ser, Pro, Tyr, Asn, or Thr”

Modified-site

/note= “Xaa at position 61 is Phe,
Asn, Glu, Pro, Lys, Arg, or Ser”

Modified-site

/note= “Xaa at position 62 is Asn,
His, Val, Arg, Pro, Thr, Asp, or Ile”

Modified-site

/note= “Xaa at position 63 is Arg,
Tyr, Trp, Lys, Ser, His, Pro, or Val”

Modified-site

/note= “Xaa at position 64 is Ala,
Asn, Pro, Ser, or Lys”

Modified-site

/note= “Xaa at position 65 is Val,
Thr, Pro, His, Leu, Phe, or Ser”

Modified-site

/note= “Xaa at position 66 is Lys,
Ile, Arg, Val, Asn, Glu, or Ser”

Modified-site

/note= “Xaa at position 67 is Ser,
Ala, Phe, Val, Gly, Asn, Ile, Pro, or His”

Modified-site

/note= “Xaa at position 68 is Leu,
Val, Trp, Ser, Ile, Phe, Thr, or His”

Modified-site

/note= “Xaa at position 69 is Gln,
Ala, Pro, Thr, Glu, Arg, Trp, Gly, or Leu”

Modified-site

/note= “Xaa at position 70 is Asn,
Leu, Val, Trp, Pro, or Ala”

Modified-site

/note= “Xaa at position 71 is
Ala,Met,Leu,Pro,Arg,Glu,Thr,Gln,Trp,or Asn”

Modified-site

/note= “Xaa at position 72 is Ser,
Glu, Met, Ala, His, Asn, Arg, or Asp”

Modified-site

/note= “Xaa at position 73 is Ala,
Glu, Asp, Leu, Ser, Gly, Thr, or Arg”

Modified-site

/note= “Xaa at position 74 is Ile,
Met, Thr, Pro, Arg, Gly, or Ala”

Modified-site

/note= “Xaa at position 75 is
Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser, Gln,
or Leu”

Modified-site

/note= “Xaa at position 76 is Ser,
Val, Ala, Asn, Trp, Glu, Pro, Gly, or Asp”

Modified-site

/note= “Xaa at position 77 is Ile,
Ser, Arg, Thr, or Leu”

Modified-site

/note= “Xaa at position 78 is Leu,
Ala, Ser, Glu, Phe, Gly, or Arg”

Modified-site

/note= “Xaa at position 79 is Lys, Thr,
Asn, Met, Arg, Ile, Gly, or Asp”

Modified-site

/note= “Xaa at position 80 is Asn,
Trp, Val, Gly, Thr, Leu, Glu, or Arg”

Modified-site

/note= “Xaa at position 81 is Leu,
Gln, Gly, Ala, Trp, Arg, Val, or Lys”

Modified-site

/note= “Xaa at position 82 is Leu,
Gln, Lys, Trp, Arg, Asp, Glu, Asn, His, Thr, Ser, Ala,
Tyr, Phe, Ile, Met, or Val”

Modified-site

/note= “Xaa at position 83 is Pro,
Ala, Thr, Trp, Arg, or Met”

Modified-site

/note= “Xaa at position 84 is Cys,
Glu, Gly, Arg, Met, or Val”

Modified-site

/note= “Xaa at position 85 is Leu,
Asn, Val, or Gln”

Modified-site

/note= “Xaa at position 86 is Pro,
Cys, Arg, Ala, or Lys”

Modified-site

/note= “Xaa at position 87 is Leu,
Ser, Trp, or Gly”

Modified-site

/note= “Xaa at position 88 is Ala,
Lys, Arg, Val, or Trp”

Modified-site

/note= “Xaa at position 89 is Thr,
Asp, Cys, Leu, Val, Glu, His, Asn, or Ser”

Modified-site

/note= “Xaa at position 90 is Ala,
Pro, Ser, Thr, Gly, Asp, Ile, or Met”

Modified-site

/note= “Xaa at position 91 is Ala,
Pro, Ser, Thr, Phe, Leu, Asp, or His”

Modified-site

/note= “Xaa at position 92 is Pro,
Phe, Arg, Ser, Lys, His, Ala, Gly, Ile, or Leu”

Modified-site

/note= “Xaa at position 93 is Thr,
Asp, Ser, Asn, Pro, Ala, Leu, or Arg”

Modified-site

/note= “Xaa at position 94 is Arg,
Ile, Ser, Glu, Leu, Val, Gln, Lys, His, Ala, or Pro”

Modified-site

/note= “Xaa at position 95 is His,
Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn, Lys, Ser, Ala,
Trp, Phe, Ile, or Tyr”

Modified-site

/note= “Xaa at position 96 is Pro,
Lys, Tyr, Gly, Ile, or Thr”

Modified-site

/note= “Xaa at position 97 is Ile,
Val, Lys, Ala, or Asn”

Modified-site

/note= “Xaa at position 98 is His,
Ile, Asn, Leu, Asp, Ala, Thr, Glu, Gln, Ser, Phe, Met,
Val, Lys, Arg, Tyr, or Pro”

Modified-site

/note= “Xaa at position 99 is Ile,
Leu, Arg, Asp, Val, Pro, Gln, Gly, Ser, Phe,
or His”

Modified-site

100

/note= “Xaa at position 100 is
Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln, or Pro”

Modified-site

101

/note= “Xaa at position 101 is
Asp, Pro, Met, Lys, His, Thr, Val, Tyr, Glu, Asn, Ser,
Ala, Gly, Ile, Leu, or Gln”

Modified-site

102

/note= “Xaa at position 102 is Gly,
Leu, Glu, Lys, Ser, Tyr, or Pro”

Modified-site

103

/note= “Xaa at position 103 is Asp,
or Ser”

Modified-site

104

/note= “Xaa at position 104 is
Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu, Gln, Lys, Ala,
Phe, or Gly”

Modified-site

105

/note= “Xaa at position 105 is
Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile,
Asp, or His”

Modified-site

106

/note= “Xaa at position 106 is Glu,
Ser, Ala, Lys, Thr, Ile, Gly, or Pro”

Modified-site

108

/note= “Xaa at position 108 is Arg,
Lys, Asp, Leu, Thr, Ile, Gln, His, Ser, Ala, or Pro”

Modified-site

109

/note= “Xaa at position 109 is Arg,
Thr, Pro, Glu, Tyr, Leu, Ser, or Gly”

Modified-site

110

/note= “Xaa at position 110 is Lys,
Ala, Asn, Thr, Leu, Arg, Gln, His, Glu, Ser,
or Trp”

Modified-site

111

/note= “Xaa at position 111 is Leu,
Ile, Arg, Asp, or Met”

Modified-site

112

/note= “Xaa at position 112 is Thr,
Val, Gln, Tyr, Glu, His, Ser, or Phe”

Modified-site

113

/note= “Xaa at position 113 is Phe,
Ser, Cys, His, Gly, Trp, Tyr, Asp, Lys, Leu, Ile, Val,
or Asn”

Modified-site

114

/note= “Xaa at position 114 is Tyr,
Cys, His, Ser, Trp, Arg, or Leu”

Modified-site

115

/note= “Xaa at position 115 is
Leu, Asn, Val, Pro, Arg, Ala, His, Thr, Trp, or
Met”

Modified-site

116

/note= “Xaa at position 116 is Lys,
Leu, Pro, Thr, Met, Asp, Val, Glu, Arg, Trp, Ser,
Asn, His, Ala, Tyr, Phe, Glu, or Ile”

Modified-site

117

/note= “Xaa at position 117 is Thr,
Ser, Asn, Ile, Trp, Lys, or Pro”

Modified-site

118

/note= “Xaa at position 118 is Leu,
Ser, Pro, Ala, Glu, Cys, Asp, or Tyr”

Modified-site

119

/note= “Xaa at position 119 is Glu,
Ser, Lys, Pro, Leu, Thr, Tyr, or Arg”

Modified-site

120

/note= “Xaa at position 120 is Asn,
Ala, Pro, Leu, His, Val, or Gln”

Modified-site

121

/note= “Xaa at position 121 is Ala,
Ser, Ile, Asn, Pro, Lys, Asp, or Gly”

Modified-site

122

/note= “Xaa at position 122 is
Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr,
or Cys”

Modified-site

123

/note= “Xaa at position 123 is Ala,
Met, Glu, His, Ser, Pro, Tyr, or Leu”

15
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn Cys
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
65 70 75 80
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
85 90 95
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Xaa Xaa Xaa Xaa Xaa
100 105 110
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gln Gln Thr Thr Leu
115 120 125
Ser Leu Ala Ile Phe
130

133 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- may or may not precede
the amino acid in position 1”

Modified-site

/note= “Xaa at position 17 is Ser,
Gly, Asp, Met, or Gln”

Modified-site

/note= “Xaa at position 18 is Asn,
His, Leu, Ile, Phe, Arg, or Gln”

Modified-site

/note= “Xaa at position 19 is Met,
Phe, Ile, Arg, or Ala”

Modified-site

/note= “Xaa at position 20 is Ile
or Pro”

Modified-site

/note; “Xaa at position 21 is Asp
or Glu”

Modified-site

/note= “Xaa at position 23 is Ile,
Val, Ala, Leu, or Gly”

Modified-site

/note= “Xaa at position 24 is Ile,
Val, Phe, or Leu”

Modified-site

/note= “Xaa at position 25 is Thr,
His, Gly, Gln, Arg, Pro, or Ala”

Modified-site

/note= “Xaa at position 26 is His,
Phe, Gly, Arg, or Ala”

Modified-site

/note= “Xaa at position 28 is Lys,
Leu, Gln, Gly, Pro, or Val”

Modified-site

/note= “Xaa at position 29 is Gln,
Asn, Leu, Arg, or Val”

Modified-site

/note= “Xaa at position 30 is Pro,
His, Thr, Gly, or Gln”

Modified-site

/note= “Xaa at position 31 is Pro,
Asp, Gly, Ala, Arg, Leu, or Gln”

Modified-site

/note= “Xaa at position 32 Leu,
Arg, Gln, Asn, Gly, Ala, or Glu”

Modified-site

/note= “Xaa at position 33 is Pro,
Leu, Gln, Ala, or Glu”

Modified-site

/note= “Xaa at position 34 is Leu,
Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe,
Thr, or Met”

Modified-site

/note= “Xaa at position 35 is Leu,
Ala, Asn, Pro, Gln, or Val”

Modified-site

/note= “Xaa at position 36 is Asp
or Leu”

Modified-site

/note= “Xaa at position 37 is Phe,
Ser, Pro, Trp, or Ile”

Modified-site

/note= “Xaa at position 38 is Asn
or Ala”

Modified-site

/note= “Xaa at position 41 is Asn,
Cys, Arg, His, Met, or Pro”

Modified-site

/note= “Xaa at position 42 is Gly,
Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr, Val,
or Arg”

Modified-site

/note=“Xaa at position 44 is Asp or
Glu”

Modified-site

/note= “Xaa at position 45 is Gln,
Val, Met, Leu, Thr, Lys, Ala, Asn, Glu, Ser, or Trp”

Modified-site

/note= “Xaa at position 46 is Asp,
Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu, His, Ile, Lys,
Tyr, Val, or Gly”

Modified-site

/note= “Xaa at position 47 is Ile, Val,
or His”

Modified-site

/note= “Xaa at position 49 is Met,
Asn, or Asp”

Modified-site

/note= “Xaa at position 50 is Glu,
Thr, Ala, Asn, Ser, or Asp”

Modified-site

/note= “Xaa at position 51 is Asn,
Arg, Met, Pro, Ser, Thr, or His”

Modified-site

/note= “Xaa at position 52 is Asn
or Gly”

Modified-site

/note= “Xaa at position 53 is Leu,
Met, or Phe”

Modified-site

/note= “Xaa at position 54 is Arg
Ala, or Ser”

Modified-site

/note= “Xaa at position 55 is Arg,
Thr, Val, Leu, or Gly”

Modified-site

/note= “Xaa at position 56 is Pro,
Gly, Cys, Ser, Gln, Ala, Arg, Asn, Glu, His, Leu, Thr,
Val, or Lys”

Modified-site

/note= “Xaa at position 59 is Glu,
Tyr, His, Leu, or Arg”

Modified-site

/note= “Xaa at position 60 is Ala,
Ser, Asn, or Thr”

Modified-site

/note= “Xaa at position 61 is Phe
or Ser”

Modified-site

/note= “Xaa at position 62 is Asn,
Val, Pro, Thr, or Ile”

Modified-site

/note= “Xaa at position 63 is Arg,
Tyr, Lys, Ser, His, or Val”

Modified-site

/note= “Xaa at position 64 is Ala
or Asn”

Modified-site

/note= “Xaa at position 65 is Val,
Thr, Leu, or Ser”

Modified-site

/note= “Xaa at position 66 is Lys,
Ile, Arg, Val, Asn, Glu, or Ser”

Modified-site

/note= “Xaa at position 67 is Ser,
Phe, Val, Gly, Asn, Ile, or His”

Modified-site

/note= “Xaa at position 68 is Leu,
Val, Ile, Phe, or His”

Modified-site

/note= “Xaa at position 69 is Gln,
Ala, Pro, Thr, Glu, Arg, or Gly”

Modified-site

/note= “Xaa at position 70 is Asn
or Pro”

Modified-site

/note= “Xaa at position 71 is Ala,
Met, Pro, Arg, Glu, Thr, or Gln”

Modified-site

/note= “Xaa at position 72 is Ser,
Glu, Met, Ala, His, Asn, Arg, or Asp”

Modified-site

/note= “Xaa at position 73 is Ala,
Glu, Asp, Leu, Ser, Gly, Thr, Arg, or Pro”

Modified-site

/note= “Xaa at position 74 is Ile
or Met”

Modified-site

/note= “Xaa at position 75 is Glu,
Gly, Asp, Ser, or Gln”

Modified-site

/note= “Xaa at position 76 is Ser,
Val, Ala, Asn, Glu, Pro, Gly, or Asp”

Modified-site

/note= “Xaa at position 77 is Ile,
Ser, or Leu”

Modified-site

/note= “Xaa at position 79 is Lys,
Thr, Asn, Met, Arg, Ile, Gly, or Asp”

Modified-site

/note= “Xaa at position 80 is Asn,
Val, Gly, Thr, Leu, Glu, or Arg”

Modified-site

/note= “Xaa at position 81 is Leu
or Val”

Modified-site

/note= “Xaa at position 82 is Leu,
Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Met, Phe,
Ser, Thr, Tyr, or Val”

Modified-site

/note= “Xaa at position 83 is Pro,
Ala, Thr, Trp, or Met”

Modified-site

/note= “Xaa at position 85 is Leu
or Val”

Modified-site

/note= “Xaa at position 87 is Leu
or Ser”

Modified-site

/note= “Xaa at position 88 is Ala,
Arg, or Trp”

Modified-site

/note= “Xaa at position 89 is Thr,
Asp, Glu, His, Asn, or Ser”

Modified-site

/note= “Xaa at position 90 is Ala,
Asp, or Met”

Modified-site

/note= “Xaa at position 91 is Ala,
Pro, Ser, Thr, Phe, Leu, or Asp”

Modified-site

/note= “Xaa at position 92 is Pro
or Ser”

Modified-site

/note= “Xaa at position 93 is Thr,
Asp, Ser, Pro, Ala, Leu, or Arg”

Modified-site

/note= “Xaa at position 95 is His,
Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe, Ser,
or Thr”

Modified-site

/note= “Xaa at position 96 is Pro
or Tyr”

Modified-site

/note= “Xaa at position 97 is Ile,
Val, or Ala”

Modified-site

/note= “Xaa at position 98 is His,
Ile, Asn, Leu, Asp, Ala, Thr, Arg, Gln, Glu,
Lys, Met, Ser, Tyr, Val, or Pro”

Modified-site

/note= “Xaa at position 99 is Ile,
Leu, Val, or Phe”

Modified-site

100

/note= “Xaa at position 100 is Lys,
Leu, His, Arg, Ile, Gln, Pro, or Ser”

Modified-site

101

/note= “Xaa at position 101 is Asp,
Pro, Met, Lys, His, Thr, Val, Asn, Ile, Leu, or Tyr”

Modified-site

102

/note= “Xaa at position 102 is Gly,
Glu, Lys, or Ser”

Modified-site

104

/note= “Xaa at position 104 is Trp,
Val, Tyr, Met, or Leu”

Modified-site

105

/note= “Xaa at position 105 is
Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys,
Ile, Asp or His”

Modified-site

106

/note= “Xaa at position 106 is Glu,
Ser, Ala, or Gly”

Modified-site

108

/note= “Xaa at position 108 is Arg,
Ala, Gln, Ser, or Lys”

Modified-site

109

/note= “Xaa at position 109 is Arg,
Thr, Glu, Leu, Ser, or Gly”

Modified-site

112

/note= “Xaa at position 112 is Thr,
Val, Gln, Glu, His, or Ser”

Modified-site

114

/note= “Xaa at position 114 is Tyr
or Trp”

Modified-site

115

/note= “Xaa at position 115 is Leu
or Ala”

Modified-site

116

/note= “Xaa at position 116 is Lys,
Thr, Met, Val, Trp, Ser, Leu, Ala, Asn, Gln, His, Met,
Phe, Tyr, or Ile”

Modified-site

117

/note= “Xaa at position 117 is Thr,
Ser, or Asn”

Modified-site

119

/note= “Xaa at position 119 is
Glu, Ser, Pro, Leu, Thr, or Tyr”

Modified-site

120

/note= “Xaa at position 120 is Asn,
Pro, Leu, His, Val, or Gln”

Modified-site

121

/note= “Xaa at position 121 is Ala,
Ser, Ile, Asn, Pro, Lys, Asp, or Gly”

Modified-site

122

/note= “Xaa at position 122 is
Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr,
or Cys”

Modified-site

123

/note= “Xaa at position 123 is Ala,
Met, Glu, His, Ser, Pro, Tyr, or Leu”

16
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn Cys
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Xaa Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa
65 70 75 80
Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa Xaa Xaa Xaa Xaa Arg Xaa Xaa
85 90 95
Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Xaa Phe Xaa Xaa Lys Leu Xaa
100 105 110
Phe Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Gln Gln Thr Thr Leu
115 120 125
Ser Leu Ala Ile Phe
130

133 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- may or may not precede
the amino acid in position 1”

Modified-site

/note= “Xaa at position 17 is Ser,
Gly, Asp, Met, or Gln”

Modified-site

/note= “Xaa at position 18 is Asn,
His, or Ile”

Modified-site

/note= “Xaa at position 19 is Met
or Ile”

Modified-site

/note= “Xaa at position 21 is Asp
or Glu”

Modified-site

/note= “Xaa at position 23 is Ile,
Ala, Leu, or Gly”

Modified-site

/note= “Xaa at position 24 is Ile,
Val, or Leu”

Modified-site

/note= “Xaa at position 25 is Thr,
His, Gln, or Ala”

Modified-site

/note= “Xaa at position 26 is His
or Ala”

Modified-site

/note= “Xaa at position 29 is Gln,
Asn, or Val”

Modified-site

/note= “Xaa at position 30 is Pro,
Gly, or Gln”

Modified-site

/note= “Xaa at position 31 is Pro,
Asp, Gly, or Gln”

Modified-site

/note= “Xaa at position 32 is Leu,
Arg, Gln, Asn, Gly, Ala, or Glu”

Modified-site

/note= “Xaa at position 33 is Pro
or Glu”

Modified-site

/note= “Xaa at position 34 is Leu,
Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe
Thr, or Met”

Modified-site

/note= “Xaa at position 35 is Leu,
Ala, Asn, Pro, Gln, or Val”

Modified-site

/note= “Xaa at position 37 is Phe,
Ser, Pro, or Trp”

Modified-site

/note= “Xaa at position 38 is Asn
or Ala”

Modified-site

/note= “Xaa at position 42 is Gly,
Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr, or Arg”

Modified-site

/note= “Xaa at position 44 is Asp
or Glu”

Modified-site

/note= “Xaa at position 45 is Gln,
Val, Met, Leu, Thr, Ala, Asn, Glu, Ser, or Lys”

Modified-site

/note= “Xaa at position 46 is Asp,
Phe, Ser, Thr, Ala, Asn, Gln, Glu, His, Ile, Lys,
Tyr, Val, or Cys”

Modified-site

/note= “Xaa at position 50 is Glu,
Ala, Asn, Ser, or Asp”

Modified-site

/note= “Xaa at position 51 is Asn,
Arg, Met, Pro, Ser, Thr, or His”

Modified-site

/note= “Xaa at position 54 is Arg
or Ala”

Modified-site

/note= “Xaa at position 55 is Arg,
Thr, Val, Leu, or Gly”

Modified-site

/note= “Xaa at position 56 is Pro,
Gly, Ser, Gln, Ala, Arg, Asn, Glu, Leu, Thr, Val,
or Lys”

Modified-site

/note= “Xaa at position 60 is Ala
or Ser”

Modified-site

/note= “Xaa at position 62 is Asn,
Pro, Thr, or Ile”

Modified-site

/note= “Xaa at position 63 is Arg
or Lys”

Modified-site

/note= “Xaa at position 64 is Ala
or Asn”

Modified-site

/note= “Xaa at position 65 is Val
or Thr”

Modified-site

/note= “Xaa at position 66 is Lys
or Arg”

Modified-site

/note= “Xaa at position 67 is Ser
Phe or His”

Modified-site

/note= “Xaa at position 68 is Leu,
Ile, Phe, or His”

Modified-site

/note= “Xaa at position 69 is Gln,
Ala, Pro, Thr, Glu, Arg, or Gly”

Modified-site

/note= “Xaa at position 71 is Ala,
Pro, or Arg”

Modified-site

/note= “Xaa at position 72 is Ser,
Glu, Arg, or Asp”

Modified-site

/note= “Xaa at position 73 is Ala
or Leu”

Modified-site

/note= “Xaa at position 76 is Ser,
Val, Ala, Asn, Glu, Pro, or Gly”

Modified-site

/note= “Xaa at position 77 is Ile
or Leu”

Modified-site

/note= “Xaa at position 79 is
Lys, Thr, Asn, Met, Arg, Ile, Gly, or Asp”

Modified-site

/note= “Xaa at position 80 is Asn,
Gly, Glu, or Arg”

Modified-site

/note= “Xaa at position 82 is Leu,
Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Ile, Met,
Phe, Ser, Thr, Tyr, or Val”

Modified-site

/note= “Xaa at position 83 is Pro
or Thr”

Modified-site

/note= “Xaa at position 85 is Leu
or Val”

Modified-site

/note= “Xaa at position 87 is Leu
or Ser”

Modified-site

/note= “Xaa at position 88 is Ala
or Trp”

Modified-site

/note= “Xaa at position 91 is Ala
or Pro”

Modified-site

/note= “Xaa at position 93 is Thr,
Asp, Ser, Pro, Ala, Leu, or Arg”

Modified-site

/note= “Xaa at position 95 is His,
Pro, Arg, Val, Leu, Gly, Asn, Phe, Ser, or Thr”

Modified-site

/note= “Xaa at position 96 is Pro
or Tyr”

Modified-site

/note= “Xaa at position 97 is Ile
or Val”

Modified-site

/note= “Xaa at position 98 is His,
Ile, Asn, Leu, Ala, Thr, Arg, Gln, Lys,
Met, Ser, Tyr, Val, or Pro”

Modified-site

/note= “Xaa at position 99 is Ile,
Leu, or Val”

Modified-site

100

/note= “Xaa at position 100 is Lys,
Arg, Ile, Gln, Pro, or Ser”

Modified-site

101

/note= “Xaa at position 101 is Asp,
Pro, Met, Lys, His, Thr, Pro, Asn, Ile, Leu, or Tyr”

Modified-site

104

/note= “Xaa at position 104 is Trp
or Leu”

Modified-site

105

/note= “Xaa at position 105 is
Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu, Lys, Ile,
Asp, or His”

Modified-site

106

/note= “Xaa at position 106 is Glu
or Gly”

Modified-site

109

/note= “Xaa at position 109 is Arg,
Thr, Glu, Leu, or Ser”

Modified-site

112

/note= “Xaa at position 112 is Thr,
Val, or Gln”

Modified-site

114

/note= “Xaa at position 114 is Tyr
or Trp”

Modified-site

115

/note= “Xaa at position 115 is Leu
or Ala”

Modified-site

116

/note= “Xaa at position 116 is Lys,
Thr, Val, Trp, Ser, Ala, His, Met, Phe, Tyr, or Ile”

Modified-site

117

/note= “Xaa at position 117 is Thr
or Ser”

Modified-site

120

/note= “Xaa at position 120 is Asn,
Pro, Leu, His, Val, or Gln”

Modified-site

121

/note= “Xaa at position 121 is Ala,
Ser, Ile, Asn, Pro, Asp, or Gly”

Modified-site

122

/note= “Xaa at position 122 is
Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr,
or Cys”

Modified-site

123

/note= “Xaa at position 123 is Ala,
Met, Glu, His, Ser, Pro, Tyr, or Leu”

17
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn Cys
1 5 10 15
Xaa Xaa Xaa Ile Xaa Glu Xaa Xaa Xaa Xaa Leu Lys Xaa Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa Xaa Xaa Ile Leu
35 40 45
Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Glu Xaa Phe Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile Glu Xaa Xaa Leu Xaa Xaa
65 70 75 80
Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr Ala Xaa Pro Xaa Arg Xaa Xaa
85 90 95
Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa Xaa Xaa Phe Xaa Xaa Lys Leu Xaa
100 105 110
Phe Xaa Xaa Xaa Xaa Leu Glu Xaa Xaa Xaa Xaa Gln Gln Thr Thr Leu
115 120 125
Ser Leu Ala Ile Phe
130

133 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- may or may not precede
the amino acid in position 1”

Modified-site

/note= “Xaa at position 17 is Ser,
Gly, Asp, or Gln”

Modified-site

/note= “Xaa at position 18 is Asn,
His, or Ile”

Modified-site

/note= “Xaa at position 23 is Ile,
Ala, Leu, or Gly”

Modified-site

/note= “Xaa at position 25 is Thr,
His, or Gln”

Modified-site

/note= “Xaa at position 26 is His
or Ala”

Modified-site

/note=“Xaa at position 29 is Gln
or Asn”

Modified-site

/note= “Xaa at position 30 is Pro
or Gly”

Modified-site

/note= “Xaa at position 32 is Leu,
Arg, Asn, or Ala”

Modified-site

/note= “Xaa at position 34 is Leu,
Val, Ser, Ala, Arg, Gln, Glu, Ile, Phe, Thr, or Met”

Modified-site

/note= “Xaa at position 35 is Leu,
Ala, Asn, or Pro”

Modified-site

/note= “Xaa at position 38 is Asn
or Ala”

Modified-site

/note= “Xaa at position 42 is Gly,
Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyr, or Arg”

Modified-site

/note= “Xaa at position 45 is Gln,
Val, Met, Leu, Ala, Asn, Glu, or Lys”

Modified-site

/note= “Xaa at position 46 is Asp,
Phe, Ser, Gln, Glu, His, Val, or Thr”

Modified-site

/note= “Xaa at position 50 is Glu,
Asn, Ser, or Asp”

Modified-site

/note= “Xaa at position 51 is Asn,
Arg, Pro, Thr, or His”

Modified-site

/note= “Xaa at position 55 is Arg,
Leu, or Gly”

Modified-site

/note= “Xaa at position 56 is Pro,
Gly, Ser, Ala, Asn, Val, Leu, or Gln”

Modified-site

/note= “Xaa at position 62 is Asn,
Pro, or Thr”

Modified-site

/note= “Xaa at position 64 is Ala
or Asn”

Modified-site

/note= “Xaa at position 65 is Val
or Thr”

Modified-site

/note= “Xaa at position 67 is Ser
or Phe”

Modified-site

/note= “Xaa at position 68 is Leu
or Phe”

Modified-site

/note= “Xaa at position 69 is Gln,
Ala, Glu, or Arg”

Modified-site

/note= “Xaa at position 76 is Ser,
Val, Asn, Pro, or Gly”

Modified-site

/note= “Xaa at position 77 is Ile
or Leu”

Modified-site

/note= “Xaa at position 79 is Lys,
Asn, Met, Arg, Ile, or Gly”

Modified-site

/note= “Xaa at position 80 is Asn,
Gly, Glu, or Arg”

Modified-site

/note= “Xaa at position 82 is Leu,
Gln, Trp, Arg, Asp, Asn, Glu, His, Met, Phe, Ser,
Thr, Tyr, or Val”

Modified-site

/note= “Xaa at position 87 is Leu
or Ser”

Modified-site

/note= “Xaa at position 88 is Ala
or Trp”

Modified-site

/note= “Xaa at position 91 is Ala
or Pro”

Modified-site

/note= “Xaa at position 93 is Thr,
Asp, or Ala”

Modified-site

/note= “Xaa at position 95 is His,
Pro, Arg, Val, Gly, Asn, Ser, or Thr”

Modified-site

/note= “Xaa at position 98 is His,
Ile, Asn, Ala, Thr, Arg, Gln, Glu, Lys, Met, Ser,
Tyr, Val, or Leu”

Modified-site

/note= “Xaa at position 99 is Ile
or Leu”

Modified-site

100

/note= “Xaa at position 100 is Lys
or Arg”

Modified-site

101

/note= “Xaa at position 101 is Asp,
Pro, Met, Lys, Thr, His, Pro, Asn, Ile, Leu, or Tyr”

Modified-site

105

/note= “Xaa at position 105 is Asn,
Pro, Ser, Ile, or Asp”

Modified-site

108

/note= “Xaa at position 108 is Arg,
Ala, or Ser”

Modified-site

109

/note= “Xaa at position 109 is Arg,
Thr, Glu, Leu, or Ser”

Modified-site

112

/note= “Xaa at position 112 is Thr
or Gln”

Modified-site

116

/note= “Xaa at position 116 is Lys,
Val, Trp, Ala, His, Phe, Tyr, or Ile”

Modified-site

117

/note= “Xaa at position 117 is Thr
or Ser”

Modified-site

120

/note= “Xaa at position 120 is Asn,
Pro, Leu, His, Val, or Gln”

Modified-site

121

/note= “Xaa at position 121 is Ala,
Ser, Ile, Pro, or Asp”

Modified-site

122

/note= “Xaa at position 122 is Gln,
Met, Trp, Phe, Pro, His, Ile, or Tyr”

Modified-site

123

/note= “Xaa at position 123 is Ala,
Met, Glu, Ser, or Leu”

18
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn Cys
1 5 10 15
Xaa Xaa Met Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa Xaa Pro Xaa
20 25 30
Pro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Glu Asp Xaa Xaa Ile Leu
35 40 45
Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Glu Ala Phe Xaa Arg Xaa
50 55 60
Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile Glu Xaa Xaa Leu Xaa Xaa
65 70 75 80
Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr Ala Xaa Pro Xaa Arg Xaa Pro
85 90 95
Ile Xaa Xaa Xaa Xaa Gly Asp Trp Xaa Glu Phe Xaa Xaa Lys Leu Xaa
100 105 110
Phe Tyr Leu Xaa Xaa Leu Glu Xaa Xaa Xaa Xaa Gln Gln Thr Thr Leu
115 120 125
Ser Leu Ala Ile Phe
130

111 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- or Met-Ala- may or may
not precede the amino acid in position 1”

Modified-site

/note= “Xaa at position 3 is Ser,
Lys, Gly, Asp, Met, Gln, or Arg”

Modified-site

/note= “Xaa at position 4 is Asn,
His, Leu, Ile, Phe, Arg, or Gln”

Modified-site

/note= “Xaa at position 5 is Met,
Phe, Ile, Arg, Gly, Ala, or Cys”

Modified-site

/note= “Xaa at position 6 is Ile,
Cys, Gln, Glu, Arg, Pro, or Ala”

Modified-site

/note= “Xaa at position 7 is Asp,
Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn, Thr, Ser,
or Val”

Modified-site

/note= “Xaa at position 8 is Glu,
Trp, Pro, Ser, Ala, His, Asp, Asn, Gln, Leu, Val,
or Gly”

Modified-site

/note= “Xaa at position 9 is
Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe, Ser
or Arg”

Modified-site

/note= “Xaa at position 10 is Ile,
Gly, Val, Arg, Ser, Phe, or Leu”

Modified-site

/note= “Xaa at position 11 is Thr,
His, Gly, Gln, Arg, Pro, or Ala”

Modified-site

/note= “Xaa at position 12 is His,
Thr, Phe, Gly, Arg, Ala, or Trp”

Modified-site

/note= “Xaa at position 13 is Leu,
Gly, Arg, Thr, Ser, or Ala”

Modified-site

/note= “Xaa at position 14 is Lys,
Arg, Leu, Gln, Gly, Pro, Val, or Trp”

Modified-site

/note= “Xaa at position 15 is Gln,
Asn, Leu, Pro, Arg, or Val”

Modified-site

/note= “Xaa at position 16 is Pro,
His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys”

Modified-site

/note= “Xaa at position 17 is Pro,
Asp, Gly, Ala, Arg, Leu, or Gln”

Modified-site

/note= “Xaa at position 18 is Leu,
Val, Arg, Gln, Asn, Gly, Ala, or Glu”

Modified-site

/note= “Xaa at position 19 is Pro,
Leu, Gln, Ala, Thr, or Glu”

Modified-site

/note= “Xaa at position 20 is Leu,
Val, Gly, Ser, Lys, Glu, Gln, Thr, Arg, Ala, Phe,
Ile, or Met”

Modified-site

/note= “Xaa at position 21 is Leu,
Ala, Gly, Asn, Pro, Gln, or Val”

Modified-site

/note= “Xaa at position 22 is Asp,
Leu, or Val”

Modified-site

/note= “Xaa at position 23 is Phe,
Ser, Pro, Trp, or Ile”

Modified-site

/note= “Xaa at position 24 is Asn
or Ala”

Modified-site

/note= “Xaa at position 26 is Leu,
Trp, or Arg”

Modified-site

/note= “Xaa at position 27 is Asn,
Cys, Arg, Leu, His, Met, or Pro”

Modified-site

/note= “Xaa at position 28 is Gly,
Asp, Ser, Cys, Ala, Lys, Asn, Thr, Leu, Val, Glu,
Phe, Tyr, Ile, or Met”

Modified-site

/note= “Xaa at position 29 is Glu,
Asn, Tyr, Leu, Phe, Asp, Ala, Cys, Gln, Arg, Thr,
Gly, or Ser”

Modified-site

/note= “Xaa at position 30 is Asp,
Ser, Leu, Arg, Lys, Thr, Met, Trp, Glu, Asn, Gln,
Ala, or Pro”

Modified-site

/note= “Xaa at position 31 is Gln,
Pro, Phe, Val, Met, Leu, Thr, Lys, Asp, Asn, Arg,
Ser, Ala, Ile, Glu, His, or Trp”

Modified-site

/note= “Xaa at position 32 is Asp,
Phe, Ser, Thr, Cys, Glu, Asn, Gln, Lys, His, Ala,
Tyr, Ile, Val, or Gly”

Modified-site

/note= “Xaa at position 33 is Ile,
Gly, Val, Ser, Arg, Pro, or His”

Modified-site

/note= “Xaa at position 34 is Leu,
Ser, Cys, Arg, Ile, His, Phe, Glu, Lys, Thr, Ala,
Met, Val, or Asn”

Modified-site

/note= “Xaa at position 35 is Met,
Arg, Ala, Gly, Pro, Asn, His, or Asp”

Modified-site

/note= “Xaa at position 36 is Glu,
Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala, Ile, Val,
His, Phe, Met, or Gln”

Modified-site

/note= “Xaa at position 37 is Asn,
Arg, Met, Pro, Ser, Thr, or His”

Modified-site

/note= “Xaa at position 38 is Asn,
His, Arg, Leu, Gly, Ser, or Thr”

Modified-site

/note= “Xaa at position 39 is
Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, or Met”

Modified-site

/note= “Xaa at position 40 is Arg,
Asp, Ile, Ser, Val, Thr, Gln, Asn, Lys, His,
Ala, or Leu”

Modified-site

/note= “Xaa at position 41 is Arg,
Thr, Val, Ser, Leu, or Gly”

Modified-site

/note= “Xaa at position 42 is Pro,
Gly, Cys, Ser, Gln, Glu, Arg, His, Thr, Ala, Tyr,
Phe, Leu, Val, or Lys”

Modified-site

/note= “Xaa at position 43 is Asn
or Gly”

Modified-site

/note= “Xaa at position 44 is Leu,
Ser, Asp, Arg, Gln, Val, or Cys”

Modified-site

/note= “Xaa at position 45 is Glu,
Tyr, His, Leu, Pro, or Arg”

Modified-site

/note= “Xaa at position 46 is Ala,
Ser, Pro, Tyr, Asn, or Thr”

Modified-site

/note= “Xaa at position 47 is Phe,
Asn, Glu, Pro, Lys, Arg, or Ser”

Modified-site

/note= “Xaa at position 48 is Asn,
His, Val, Arg, Pro, Thr, Asp, or Ile”

Modified-site

/note= “Xaa at position 49 is Arg,
Tyr, Trp, Lys, Ser, His, Pro, or Val”

Modified-site

/note= “Xaa at position 50 is Ala,
Asn, Pro, Ser, or Lys”

Modified-site

/note= “Xaa at position 51 is Val,
Thr, Pro, His, Leu, Phe, or Ser”

Modified-site

/note= “Xaa at position 52 is Lys,
Ile, Arg, Val, Asn, Glu, or Ser”

Modified-site

/note= “Xaa at position 53 is Ser,
Ala, Phe, Val, Gly, Asn, Ile, Pro, or His”

Modified-site

/note= “Xaa at position 54 is Leu,
Val, Trp, Ser, Ile, Phe, Thr, or His”

Modified-site

/note= “Xaa at position 55 is Gln,
Ala, Pro, Thr, Glu, Arg, Trp, Gly, or Leu”

Modified-site

/note= “Xaa at position 56 is Asn,
Leu, Val, Trp, Pro, or Ala”

Modified-site

/note= “Xaa at position 57 is Ala,
Met, Leu, Pro, Arg, Glu, Thr, Gln, Trp, or Asn”

Modified-site

/note= “Xaa at position 58 is Ser,
Glu, Met, Ala, His, Asn, Arg, or Asp”

Modified-site

/note= “Xaa at position 59 is Ala,
Glu, Asp, Leu, Ser, Gly, Thr, or Arg”

Modified-site

/note= “Xaa at position 60 is Ile,
Met, Thr, Pro, Arg, Gly, Ala”

Modified-site

/note= “Xaa at position 61 is
Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser, Gln,
or Leu”

Modified-site

/note= “Xaa at position 62 is Ser,
Val, Ala, Asn, Trp, Glu, Pro, Gly, or Asp”

Modified-site

/note= “Xaa at position 63 is Ile,
Ser, Arg, Thr, or Leu”

Modified-site

/note= “Xaa at position 64 is Leu,
Ala, Ser, Glu, Phe, Gly, or Arg”

Modified-site

/note= “Xaa at position 65 is Lys,
Thr, Gly, Asn, Met, Arg, Ile, or Asp”

Modified-site

/note= “Xaa at position 66 is Asn,
Trp, Val, Gly, Thr, Leu, Glu, or Arg”

Modified-site

/note= “Xaa at position 67 is Leu,
Gln, Gly, Ala, Trp, Arg, Val, or Lys”

Modified-site

/note= “Xaa at position 68 is Leu,
Gln, Lys, Trp, Arg, Asp, Glu, Asn, His, Thr, Ser, Ala,
Tyr, Phe, Ile, Met, or Val”

Modified-site

/note= “Xaa at position 69 is Pro,
Ala, Thr, Trp, Arg, or Met”

Modified-site

/note= “Xaa at position 70 is Cys,
Glu, Gly, Arg, Met, or Val”

Modified-site

/note= “Xaa at position 71 is Leu,
Asn, Val, or Gln”

Modified-site

/note= “Xaa at position 72 is Pro,
Cys, Arg, Ala, or Lys”

Modified-site

/note= “Xaa at position 73 is Leu,
Ser, Trp, or Gly”

Modified-site

/note= “Xaa at position 74 is Ala,
Lys, Arg, Val, or Trp”

Modified-site

/note= “Xaa at position 75 is Thr,
Asp, Cys, Leu, Val, Glu, His, Asn, or Ser”

Modified-site

/note= “Xaa at position 76 is Ala,
Pro, Ser, Thr, Gly, Asp, Ile, or Met”

Modified-site

/note= “Xaa at position 77 is Ala,
Pro, Ser, Thr, Phe, Leu, Asp, or His”

Modified-site

/note= “Xaa at position 78 is Pro,
Phe, Arg, Ser, Lys, His, Ala, Gly, Ile, or Leu”

Modified-site

/note= “Xaa at position 79 is Thr,
Asp, Ser, Asn, Pro, Ala, Leu, or Arg”

Modified-site

/note= “Xaa at position 80 is Arg,
Ile, Ser, Glu, Leu, Val, Gln, Lys, His, Ala, or Pro”

Modified-site

/note= “Xaa at position 81 is His,
Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn, Lys, Ser,
Ala, Trp, Phe, Ile, or Tyr”

Modified-site

/note= “Xaa at position 82 is Pro,
Lys, Tyr, Gly, Ile, or Thr”

Modified-site

/note= “Xaa at position 83 is Ile,
Val, Lys, Ala, or Asn”

Modified-site

/note= “Xaa at position 84 is His,
Ile, Asn, Leu, Asp, Ala, Thr, Glu, Gln, Ser,
Phe, Met, Val, Lys, Arg, Tyr, or Pro”

Modified-site

/note= “Xaa at position 85 is
Ile, Leu, Arg, Asp, Val, Pro, Gln, Gly, Ser,
Phe, or His”

Modified-site

/note= “Xaa at position 86 is
Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln, or Pro”

Modified-site

/note= “Xaa at position 87 is
Asp, Pro, Met, Lys, His, Thr, Val, Tyr, Glu, Asn,
Ser, Ala, Gly, Ile, Leu, or Gln”

Modified-site

/note= “Xaa at position 88 Gly,
Leu, Glu, Lys, Ser, Tyr, or Pro”

Modified-site

/note= “Xaa at position 89 is Asp
or Ser”

Modified-site

/note= “Xaa at position 90 is
Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu, Gln, Lys,
Ala, Phe, or Gly”

Modified-site

/note= “Xaa at position 91 is
Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys,
Ile, Asp, or His”

Modified-site

/note= “Xaa at position 92 is Glu,
Ser, Ala, Lys, Thr, Ile, Gly, or Pro”

Modified-site

/note= “Xaa at position 94 is Arg,
Lys, Asp, Leu, Thr, Ile, Gln, His, Ser, Ala, or Pro”

Modified-site

/note= “Xaa at position 95 is Arg,
Thr, Pro, Glu, Tyr, Leu, Ser, or Gly”

Modified-site

/note= “Xaa at position 96 is Lys,
Asn, Thr, Leu, Gln, Arg, His, Glu, Ser, Ala,
or Trp”

Modified-site

/note= “Xaa at position 97 is Leu,
Ile, Arg, Asp, or Met”

Modified-site

/note= “Xaa at position 98 is Thr,
Val, Gln, Tyr, Glu, His, Ser, or Phe”

Modified-site

/note= “Xaa at position 99 is Phe,
Ser, Cys, His, Gly, Trp, Tyr, Asp, Lys, Leu, Ile,
Val, or Asn”

Modified-site

100

/note= “Xaa at position 100 is Tyr,
Cys, His, Ser, Trp, Arg, or Leu”

Modified-site

101

/note= “Xaa at position 101 is Leu,
Asn, Val, Pro, Arg, Ala, His, Thr, Trp, or Met”

Modified-site

102

/note= “Xaa at position 102 is
Lys, Leu, Pro, Thr, Met, Asp, Val, Glu, Arg, Trp,
Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile”

Modified-site

103

/note= “Xaa at position 103 is Thr,
Ser, Asn, Ile, Trp, Lys, or Pro”

Modified-site

104

/note= “Xaa at position 104 is Leu,
Ser, Pro, Ala, Glu, Cys, Asp, or Tyr”

Modified-site

105

/note= “Xaa at position 105 is Glu,
Ser, Lys, Pro, Leu, Thr, Tyr, or Arg”

Modified-site

106

/note= “Xaa at position 106 is Asn,
Ala, Pro, Leu, His, Val or Gln”

Modified-site

107

/note= “Xaa at position 107 is Ala,
Ser, Ile, Asn, Pro, Lys, Asp, or Gly”

Modified-site

108

/note= “Xaa at position 108 is
Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr,
or Cys”

Modified-site

109

/note= “Xaa at position 109 is Ala,
Met, Glu, His, Ser, Pro, Tyr, or Leu”

19
Asn Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa
20 25 30
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
50 55 60
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
65 70 75 80
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Xaa Xaa Xaa
85 90 95
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- or Met-Ala- may or may
not precede the amino acid in position 1”

Modified-site

/note= “Xaa at position 3 is Ser,
Gly, Asp, Met, or Gln”

Modified-site

/note= “Xaa at position 4 is Asn,
His, Leu,Ile, Phe, Arg, or Gln”

Modified-site

/note= “Xaa at position 5 is Met,
Phe, Ile, Arg, or Ala”

Modified-site

/note= “Xaa at position 6 is Ile or
Pro”

Modified-site

/note= “Xaa at position 7 is Asp or
Glu”

Modified-site

/note= “Xaa at position 9 is Ile,
Val, Ala, Leu, or Gly”

Modified-site

/note= “Xaa at position 10 is Ile,
Val, Phe, or Leu”

Modified-site

/note= “Xaa at position 11 is Thr,
His, Gly, Gln, Arg, Pro, or Ala”

Modified-site

/note= “Xaa at position 12 is His,
Phe, Gly, Arg, or Ala”

Modified-site

/note= “Xaa at position 14 is Lys,
Leu, Gln, Gly, Pro, or Val”

Modified-site

/note= “Xaa at position 15 is Gln,
Asn, Leu, Arg, or Val”

Modified-site

/note= “Xaa at position 16 is Pro,
His, Thr, Gly, or Gln”

Modified-site

/note= “Xaa at position 17 is Pro,
Asp, Gly, Ala, Arg, Leu, or Gln”

Modified-site

/note= “Xaa at position 18 is Leu,
Arg, Gln, Asn, Gly, Ala or Glu”

Modified-site

/note= “Xaa at poisiton 19 is Pro,
Leu, Gln, Ala, or Glu”

Modified-site

/note= “Xaa at positon 20 is Leu,
Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe,
Thr, or Met”

Modified-site

/note= “Xaa at position 21 is Leu,
Ala, Asn, Pro, Gln, or Val”

Modified-site

/note= “Xaa at position 22 is Asp
or Leu”

Modified-site

/note= “Xaa at position 23 is Phe,
Ser, Pro, Trp, or Ile”

Modified-site

/note= “Xaa at position 24 is Asn
or Ala”

Modified-site

/note= “Xaa at position 27 is Asn,
Cys, Arg, His, Met, or Pro”

Modified-site

/note= “Xaa at position 28 is Gly,
Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr, or Arg”

Modified-site

/note= “Xaa at position 30 is Asp
or Glu”

Modified-site

/note= “Xaa at position 31 is Gln,
Val, Met, Leu, Thr, Lys, Ala, Asn, Glu, Ser, or Trp”

Modified-site

/note= “Xaa at position 32 is Asp,
Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu, His, Ile,
Lys, Tyr, Val, or Gly”

Modified-site

/note= “Xaa at position 33 is Ile,
Val, or His”

Modified-site

/note= “Xaa at position 35 is Met,
Asn, or Asp”

Modified-site

/note= “Xaa at position 36 is Glu,
Thr, Ala, Asn, Ser, or Asp”

Modified-site

/note= “Xaa at position 37 is Asn,
Arg, Met, Pro, Ser, Thr, or His”

Modified-site

/note= “Xaa at position 38 is Asn
or Gly”

Modified-site

/note= “Xaa at position 39 is Leu,
Met, or Phe”

Modified-site

/note= “Xaa at position 40 is Arg,
Ala, or Ser”

Modified-site

/note= “Xaa at position 41 is Arg,
Thr, Val, Leu, or Gly”

Modified-site

/note= “Xaa at position 42 is Pro,
Gly, Cys, Ser, Gln, Ala, Arg, Asn, Glu, His, Leu,
Thr, Val, or Lys”

Modified-site

/note= “Xaa at position 45 is Glu,
Tyr, His, Leu, or Arg”

Modified-site

/note= “Xaa at position 46 is Ala,
Ser, Asn, or Thr”

Modified-site

/note= “Xaa at position 47 is Phe
or Ser”

Modified-site

/note= “Xaa at position 48 is Asn,
Val, Pro, Thr, or Ile”

Modified-site

/note= “Xaa at position 49 is Arg,
Tyr, Lys, Ser, His, or Val”

Modified-site

/note= “Xaa at position 50 is Ala
or Asn”

Modified-site

/note= “Xaa at position 51 is Val,
Thr, Leu, or Ser”

Modified-site

/note= “Xaa at position 52 is Lys,
Ile, Arg, Val, Asn, Glu, or Ser”

Modified-site

/note= “Xaa at position 53 is Ser,
Phe, Val, Gly, Asn, Ile, or His”

Modified-site

/note= “Xaa at position 54 is Leu,
Val, Ile, Phe, or His”

Modified-site

/note= “Xaa at position 55 is Gln,
Ala, Pro, Thr, Glu, Arg, or Gly”

Modified-site

/note= “Xaa at position 56 is Asn
or Pro”

Modified-site

/note= “Xaa at position 57 is Ala,
Met, Pro, Arg, Glu, Thr, or Gln”

Modified-site

/note= “Xaa at position 58 is Ser,
Glu, Met, Ala, His, Asn, Arg, or Asp”

Modified-site

/note= “Xaa at position 59 is Ala,
Glu, Asp, Leu, Ser, Gly, Thr, Arg, or Pro”

Modified-site

/note= “Xaa at position 60 is Ile
or Met”

Modified-site

/note= “Xaa at position 61 is Glu,
Gly, Asp, Ser, or Gln”

Modified-site

/note= “Xaa at position 62 is Ser,
Val, Ala, Asn, Glu, Pro, Gly, or Asp”

Modified-site

/note= “Xaa at position 63 is Ile,
Ser, or Leu”

Modified-site

/note= “Xaa at position 65 is Lys,
Thr, Gly, Asn, Met, Arg, Ile, or Asp”

Modified-site

/note= “Xaa at position 66 is Asn,
Val, Gly, Thr, Leu, Glu, or Arg”

Modified-site

/note= “Xaa at position 67 is Leu
or Val”

Modified-site

/note= “Xaa at position 68 is Leu,
Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Met, Phe,
Ser, Thr, Tyr, or Val”

Modified-site

/note= “Xaa at position 69 is Pro,
Ala, Thr, Trp, or Met”

Modified-site

/note= “Xaa at position 71 is Leu
or Val”

Modified-site

/note= “Xaa at position 73 is Leu
or Ser”

Modified-site

/note= “Xaa at position 74 is Ala,
Arg, or Trp”

Modified-site

/note= “Xaa at position 75 is Thr,
Asp, Glu, His, Asn, or Ser”

Modified-site

/note= “Xaa at position 76 is Ala,
Asp, or Met”

Modified-site

/note= “Xaa at position 77 is Ala,
Pro, Ser, Thr, Phe, Leu, or Asp”

Modified-site

/note= “Xaa at position 78 is Pro
or Ser”

Modified-site

/note= “Xaa at position 79 is Thr,
Asp, Ser, Pro, Ala, Leu, or Arg”

Modified-site

/note= “Xaa at position 81 is His,
Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe, Ser, or Thr”

Modified-site

/note= “Xaa at position 82 is Pro
or Tyr”

Modified-site

/note= “Xaa at position 83 is Ile,
Val, or Ala”

Modified-site

/note= “Xaa at position 84 is His,
Ile, Asn, Leu, Asp, Ala, Thr, Arg, Gln, Glu, Lys,
Met, Ser, Tyr, Val, or Pro”

Modified-site

/note= “Xaa at position 85 is Ile,
Leu, Val, or Phe”

Modified-site

/note= “Xaa at position 86 is Lys,
Leu, His, Arg, Ile, Gln, Pro, or Ser”

Modified-site

/note= “Xaa at position 87 is Asp,
Pro, Met, Lys, His, Thr, Val, Asn, Ile, Leu, or Tyr”

Modified-site

/note= “Xaa at position 88 is Gly,
Glu, Lys, or Ser”

Modified-site

/note= “Xaa at position 90 is Trp,
Val, Tyr, Met, or Leu”

Modified-site

/note=
“Xaa at position 91 is Asn, Pro, Ala, Phe, Ser,
Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His”

Modified-site

/note= “Xaa at position 92 is Glu,
Ser, Ala, or Gly”

Modified-site

/note= “Xaa at position 94 is Arg,
Ala, Gln, Ser, or Lys”

Modified-site

/note= “Xaa at position 95 Arg,
Thr, Glu, Leu, Ser, or Gly”

Modified-site

/note= “Xaa at position 98 is Thr,
Val, Gln, Glu, His, or Ser”

Modified-site

100

/note= “Xaa at position 100 is Tyr
or Trp”

Modified-site

101

/note= “Xaa at position 101 is Leu
or Ala”

Modified-site

102

/note= “Xaa at position 102 is Lys,
Thr, Met, Val, Trp, Ser, Leu, Ala, Asn, Gln, His,
Met, Phe, Tyr, or Ile”

Modified-site

103

/note= “Xaa at position 103 is Thr,
Ser, or Asn”

Modified-site

105

/note= “Xaa at position 105 is Glu,
Ser, Pro, Leu, Thr, or Tyr”

Modified-site

106

/note= “Xaa at position 106 is Asn,
Pro, Leu, His, Val, or Gln”

Modified-site

107

/note= “Xaa at position 107 is Ala,
Ser, Ile, Asn, Pro, Lys, Asp, or Gly”

Modified-site

108

/note=
“Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe,
Pro, His, Ile, Tyr, or Cys”

Modified-site

109

/note= “Xaa at position 109 is Ala,
Met, Glu, His, Ser, Pro, Tyr, or Leu”

20
Asn Cys Xaa Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa Xaa Xaa
20 25 30
Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Xaa Xaa
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu
50 55 60
Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa Xaa Xaa Xaa Xaa Arg
65 70 75 80
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Xaa Phe Xaa Xaa Lys
85 90 95
Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- or Met-Ala- may or may
not precede the amino acid in position 1”

Modified-site

/note= “Xaa at position 3 is Ser,
Gly, Asp, Met, or Gln”

Modified-site

/note= “Xaa at position 4 is Asn,
His, or Ile”

Modified-site

/note= “Xaa at position 5 is Met
or Ile”

Modified-site

/note= “Xaa at position 7 is Asp or Glu”

Modified-site

/note= “Xaa at position 9 is Ile,
Ala, Leu, or Gly”

Modified-site

/note= “Xaa at position 10 is Ile,
Val, or Leu”

Modified-site

/note= “Xaa at position 11 is Thr,
His, Gln, or Ala”

Modified-site

/note= “Xaa at position 12 is His
or Ala”

Modified-site

/note= “Xaa at position 15 is Gln,
Asn, or Val”

Modified-site

/note= “Xaa at position 16 is Pro,
Gly, or Gln”

Modified-site

/note= “Xaa at position 17 is Pro,
Asp, Gly, or Gln”

Modified-site

/note= “Xaa at position 18 is Leu,
Arg, Gln, Asn, Gly, Ala, or Glu”

Modified-site

/note= “Xaa at position 19 is Pro
or Glu”

Modified-site

/note= “Xaa at position 20 is Leu,
Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe,
Thr, or Met”

Modified-site

/note= “Xaa at position 21 is Leu,
Ala, Asn, Pro, Gln, or Val”

Modified-site

/note= “Xaa at position 23 is Phe,
Ser, Pro, or Trp”

Modified-site

/note= “Xaa at position 24 is Asn
or Ala”

Modified-site

/note= “Xaa at position 28 is Gly,
Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met, Tyr, or Arg”

Modified-site

/note= “Xaa at position 30 is Asp
or Glu”

Modified-site

/note= “Xaa at position 31 is Gln,
Val, Met, Leu, Thr, Ala, Asn, Glu, Ser, or Lys”

Modified-site

/note= “Xaa at position 32 is Asp,
Phe, Ser, Thr, Ala, Asn, Gln, Glu, His, Ile, Lys,
Tyr, Val, or Cys”

Modified-site

/note= “Xaa at position 36 is Glu,
Ala, Asn, Ser, or Asp”

Modified-site

/note= “Xaa at position 37 is Asn,
Arg, Met, Pro, Ser, Thr, or His”

Modified-site

/note= “Xaa at position 40 is Arg
or Ala”

Modified-site

/note= “Xaa at position 41 is Arg,
Thr, Val, Leu, or Gly”

Modified-site

/note= “Xaa at position 42 is Pro,
Gly, Ser, Gln, Ala, Arg, Asn, Glu, Leu, Thr, Val,
or Lys”

Modified-site

/note= “Xaa at position 46 is Ala
or Ser”

Modified-site

/note= “Xaa at position 48 is Asn,
Pro, Thr, or Ile”

Modified-site

/note= “Xaa at position 49 is Arg
or Lys”

Modified-site

/note= “Xaa at position 50 is Ala
or Asn”

Modified-site

/note= “Xaa at position 51 is Val
or Thr”

Modified-site

/note= “Xaa at position 52 is Lys
or Arg”

Modified-site

/note= “Xaa at position 53 is Ser,
Phe, or His”

Modified-site

/note= “Xaa at position 54 is Leu,
Ile, Phe, or His”

Modified-site

/note= “Xaa at position 55 is Gln,
Ala, Pro, Thr, Glu, Arg, or Gly”

Modified-site

/note= “Xaa at position 57 is Ala,
Pro, or Arg”

Modified-site

/note= “Xaa at position 58 is Ser,
Glu, Arg, or Asp”

Modified-site

/note= “Xaa at position 59 is Ala
or Leu”

Modified-site

/note= “Xaa at position 62 is Ser,
Val, Ala, Asn, Glu, Pro, or Gly”

Modified-site

/note= “Xaa at position 63 is Ile
or Leu”

Modified-site

/note= “Xaa at position 65 is Lys,
Thr, Gly, Asn, Met, Arg, Ile, Gly, or Asp”

Modified-site

/note= “Xaa at position 66 is Asn,
Gly, Glu, or Arg”

Modified-site

/note= “Xaa at position 68 is Leu,
Gln, Trp, Arg, Asp, Ala, Asn, Glu, His, Ile, Met,
Phe, Ser, Thr, Tyr, or Val”

Modified-site

/note= “Xaa at position 69 is Pro
or Thr”

Modified-site

/note= “Xaa at position 71 is Leu
or Val”

Modified-site

/note= “Xaa at position 73 is Leu
or Ser”

Modified-site

/note= “Xaa at position 74 is Ala
or Trp”

Modified-site

/note= “Xaa at position 77 is Ala
or Pro”

Modified-site

/note= “Xaa at position 79 is Thr,
Asp, Ser, Pro, Ala, Leu, or Arg”

Modified-site

/note= “Xaa at position 81 is His,
Pro, Arg, Val, Leu, Gly, Asn, Phe, Ser, or Thr”

Modified-site

/note= “Xaa at position 82 is Pro
or Tyr”

Modified-site

/note= “Xaa at position 83 is Ile
or Val”

Modified-site

/note= “Xaa at position 84 is His,
Ile, Asn, Leu, Ala, Thr, Arg, Gln, Lys,
Met, Ser, Tyr, Val, or Pro”

Modified-site

/note= “Xaa at position 85 is Ile,
Leu, or Val”

Modified-site

/note= “Xaa at position 86 is Lys,
Arg, Ile, Gln, Pro, or Ser”

Modified-site

/note= “Xaa at position 87 is Asp,
Pro, Met, Lys, His, Thr, Asn, Ile, Leu, or Tyr”

Modified-site

/note= “Xaa at position 90 is Trp
or Leu”

Modified-site

/note=“Xaa at position 91 is Asn,
Pro, Ala, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp,
or His”

Modified-site

/note= “Xaa at position 92 is Glu
or Gly”

Modified-site

/note= “Xaa at position 94 is Arg,
Ala, or Ser”

Modified-site

/note= “Xaa at position 95 is Arg,
Thr, Glu, Leu, or Ser”

Modified-site

/note= “Xaa at position 98 is Thr,
Val, or Gln”

Modified-site

100

/note= “Xaa at position 100 is Tyr
or Trp”

Modified-site

101

/note= “Xaa at position 101 is Leu
or Ala”

Modified-site

102

/note= “Xaa at position 102 is Lys,
Thr, Val, Trp, Ser, Ala, His, Met, Phe, Tyr, or Ile”

Modified-site

103

/note= “Xaa at position 103 is Thr
or Ser”

Modified-site

106

/note= “Xaa at position 106 is Asn,
Pro, Leu, His, Val, or Gln”

Modified-site

107

/note= “Xaa at position 107 is Ala,
Ser, Ile, Asn, Pro, Asp, or Gly”

Modified-site

108

/note= “Xaa at position 108 is Gln,
Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys”

Modified-site

109

/note= “Xaa at position 109 is Ala,
Met, Glu, His, Ser, Pro, Tyr, or Leu”

21
Asn Cys Xaa Xaa Xaa Ile Xaa Glu Xaa Xaa Xaa Xaa Leu Lys Xaa Xaa
1 5 10 15
Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa Xaa Xaa
20 25 30
Ile Leu Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Glu Xaa Phe Xaa
35 40 45
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile Glu Xaa Xaa Leu
50 55 60
Xaa Xaa Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr Ala Xaa Pro Xaa Arg
65 70 75 80
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa Xaa Xaa Phe Xaa Xaa Lys
85 90 95
Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Glu Xaa Xaa Xaa Xaa Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- or Met-Ala- may or may
not precede the amino acid in position 1”

Modified-site

/note= “Xaa at position 3 is Ser,
Gly, Asp, or Gln”

Modified-site

/note= “Xaa at position 4 is Asn,
His, or Ile”

Modified-site

/note= “Xaa at position 9 is Ile,
Ala, Leu, or Gly”

Modified-site

/note= “Xaa at position 11 is Thr,
His, or Gln”

Modified-site

/note= “Xaa at position 12 is His
or Ala”

Modified-site

/note= “Xaa at position 15 is Gln
or Asn”

Modified-site

/note= “Xaa at position 16 is Pro
or Gly”

Modified-site

/note= “Xaa at position 18 is Leu,
Arg, Asn, or Ala”

Modified-site

/note= “Xaa at position 20 is Leu,
Val, Ser, Ala, Arg, Gln, Glu, Ile, Phe, Thr, or Met”

Modified-site

/note= “Xaa at position 21 is Leu,
Ala, Asn, or Pro”

Modified-site

/note= “Xaa at position 24 is Asn
or Ala”

Modified-site

/note= “Xaa at position 28 is Gly,
Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyr, or Arg”

Modified-site

/note= “Xaa at position 31 is Gln,
Val, Met, Leu, Ala, Asn, Glu, or Lys”

Modified-site

/note= “Xaa at position 32 is Asp,
Phe, Ser, Ala, Gln, Glu, His, Val, or Thr”

Modified-site

/note= “Xaa at position 36 is Glu,
Asn, Ser, or Asp”

Modified-site

/note= “Xaa at position 37 is Asn,
Arg, Pro, Thr, or His”

Modified-site

/note= “Xaa at position 41 is Arg,
Leu, or Gly”

Modified-site

/note= “Xaa at position 42 is Pro,
Gly, Ser, Ala, Asn, Val, Leu, or Gln”

Modified-site

/note= “Xaa at position 48 is Asn,
Pro, or Thr”

Modified-site

/note= “Xaa at position 50 is Ala
or Asn”

Modified-site

/note= “Xaa at position 51 is Val
or Thr”

Modified-site

/note= “Xaa at position 53 is Ser
or Phe”

Modified-site

/note= “Xaa at position 54 is Leu
or Phe”

Modified-site

/note= “Xaa at position 55 is Gln,
Ala, Glu, or Arg”

Modified-site

/note= “Xaa at position 62 is Ser,
Val, Asn, Pro, or Gly”

Modified-site

/note= “Xaa at position 63 is Ile
or Leu”

Modified-site

/note= “Xaa at position 65 is Lys,
Asn, Met, Arg, Ile, or Gly”

Modified-site

/note= “Xaa at position 66 is Asn,
Gly, Glu, or Arg”

Modified-site

/note= “Xaa at position 68 is Leu,
Gln, Trp, Arg, Asp, Asn, Glu, His, Met, Phe, Ser,
Thr, Tyr, or Val”

Modified-site

/note= “Xaa at position 73 is Leu
or Ser”

Modified-site

/note= “Xaa at position 74 is Ala
or Trp”

Modified-site

/note= “Xaa at position 77 is Ala
or Pro”

Modified-site

/note= “Xaa at position 79 is Thr,
Asp, or Ala”

Modified-site

/note= “Xaa at position 81 is His,
Pro, Arg, Val, Gly, Asn, Ser, or Thr”

Modified-site

/note= “Xaa at position 84 is His,
Ile, Asn, Ala, Thr, Arg, Gln, Glu, Lys, Met,
Ser, Tyr, Val, or Leu”

Modified-site

/note= “Xaa at position 85 is Ile
or Leu”

Modified-site

/note= “Xaa at position 86 is Lys
or Arg”

Modified-site

/note= “Xaa at position 87 is Asp,
Pro, Met, Lys, His, Pro, Asn, Ile, Leu, or Tyr”

Modified-site

/note= “Xaa at position 91 is Asn,
Pro, Ser, Ile, or Asp”

Modified-site

/note= “Xaa at position 94 is Arg,
Ala, or Ser”

Modified-site

/note= “Xaa at position 95 is Arg,
Thr, Glu, Leu, or Ser”

Modified-site

/note= “Xaa at position 98 is Thr
or Gln”

Modified-site

102

/note= “Xaa at position 102 is Lys,
Val, Trp, or Ile”

Modified-site

103

/note= “Xaa at position 103 is Thr,
Ala, His, Phe, Tyr, or Ser”

Modified-site

106

/note= “Xaa at position 106 is Asn,
Pro, Leu, His, Val, or Gln”

Modified-site

107

/note= “Xaa at position 107 is Ala,
Ser, Ile, Pro, or Asp”

Modified-site

108

/note= “Xaa at position 108 is Gln,
Met, Trp, Phe, Pro, His, Ile, or Tyr”

Modified-site

109

/note= “Xaa at position 109 is Ala,
Met, Glu, Ser, or Leu”

22
Asn Cys Xaa Xaa Met Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa Xaa
1 5 10 15
Pro Xaa Pro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Glu Asp Xaa Xaa
20 25 30
Ile Leu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Glu Ala Phe Xaa
35 40 45
Arg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile Glu Xaa Xaa Leu
50 55 60
Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr Ala Xaa Pro Xaa Arg
65 70 75 80
Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp Xaa Glu Phe Xaa Xaa Lys
85 90 95
Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu Xaa Xaa Xaa Xaa Gln Gln
100 105 110

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

23
AACAACCTCA ATGCTGAAGA CGTTGAT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

24
ATCAACGTCT TCAGCATT 18

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

25
AACAACCTCA ATTCTGAAGA CATGGAT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

26
ATCCATGTCT TCAGAATT 18

22 base pairs

nucleic acid

single

linear

DNA (synthetic)

27
CATGGGAACC ATATGTCAGG AT 22

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

28
ATCCTGACAT ATGGTTCC 18

16 base pairs

nucleic acid

single

linear

DNA (synthetic)

29
TGAACCATAT GTCAGG 16

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

30
AATTCCTGAC ATATGGTTCA TGCA 24

20 base pairs

nucleic acid

single

linear

DNA (synthetic)

31
AATTCGAACC ATATGTCAGA 20

20 base pairs

nucleic acid

single

linear

DNA (synthetic)

32
AGCTTCTGAC ATATGGTTCG 20

22 base pairs

nucleic acid

single

linear

DNA (synthetic)

33
ATCGAACCAT ATGTCAGATG CA 22

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

34
TCTGACATAT GGTTCGAT 18

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

35
ATCCTGATGG AACGAAACCT TCGACTTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

36
AAGTCGAAGG TTTCGTTCCA TCAGGAT 27

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

37
ATCCTGATGG AACGAAACCT TCGAACTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

38
AGTTCGAAGG TTTCGTTCCA TCAGGAT 27

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

39
CTCGCATTCG TAAGGGCTGT CAAG 24

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

40
CCTTACGAAT GCGAGCAGGT TTGG 24

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

41
GAGAGCTTCG TAAGGGCTGT CAAG 24

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

42
CCTTACGAAG CTCTCCAGGT TTGG 24

15 base pairs

nucleic acid

single

linear

DNA (synthetic)

43
CACTTAGAAA ATGCA 15

20 base pairs

nucleic acid

single

linear

DNA (synthetic)

44
TTTTCTAAGT GCTTGACAGC 20

15 base pairs

nucleic acid

single

linear

DNA (synthetic)

45
AACTTAGAAA ATGCA 15

20 base pairs

nucleic acid

single

linear

DNA (synthetic)

46
TTTTCTAAGT TCTTGACAGC 20

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

47
GGTGATTGGA TGTCGAGAGG GTGCGGCCGT GGCAGAGGGC AGACATGG 48

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

48
CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCA CCATCAAG 48

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

49
GATGATTGGA TGTCGAGAGG GTGCGGCCGT GGCAGAGGGC AGACATGG 48

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

50
CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCA TCATCAAG 48

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

51
TACGAGATTA CGAAGAAT 18

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

52
CGTAATCTCG TACCATGT 18

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

53
TTGGAGATTA CGAAGAAT 18

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

54
CGTAATCTCC AACCATGT 18

19 base pairs

nucleic acid

single

linear

DNA (synthetic)

55
TGCCTCAATA CCTGATGCA 19

21 base pairs

nucleic acid

single

linear

DNA (synthetic)

56
TCAGGTATTG AGGCAATTCT T 21

26 base pairs

nucleic acid

single

linear

DNA (synthetic)

57
AATTCTTGCC AGTCACCTGC CTTGAT 26

16 base pairs

nucleic acid

single

linear

DNA (synthetic)

58
GCAGGTGACT GGCAAG 16

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

59
AATTCCGGGA AAAACTGACG TTCTATCTGG TT 32

37 base pairs

nucleic acid

single

linear

DNA (synthetic)

60
CTCAAGGGAA ACCAGATAGA ACGTCAGTTT TTCCCGG 37

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

61
ACCCTTGAGC ACGCGCAGGA ACAACAGTAA TA 32

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

62
AGCTTATTAC TGTTGTTCCT GCGCGTG 27

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

63
ACCCTTGAGC AAGCGCAGGA ACAACAGTAA TA 32

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

64
AGCTTATTAC TGTTGTTCCT GCGCTTG 27

111 amino acids

amino acid

linear

peptide

65
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

66
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro
1 5 10 15
Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp Val Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

67
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro
1 5 10 15
Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp Met Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

68
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Val Pro
1 5 10 15
Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp Met Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

69
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala Phe Val
35 40 45
Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

70
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val
35 40 45
Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

71
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val
35 40 45
Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

72
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu
50 55 60
Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg
65 70 75 80
His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

73
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu
50 55 60
Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg
65 70 75 80
His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

74
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Glu Lys
85 90 95
Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

75
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Glu Lys
85 90 95
Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

76
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu
50 55 60
Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg
65 70 75 80
His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys
85 90 95
Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

77
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu
50 55 60
Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg
65 70 75 80
His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys
85 90 95
Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

78
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu
50 55 60
Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg
65 70 75 80
His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys
85 90 95
Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

79
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro
1 5 10 15
Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp Val Asp
20 25 30
Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val
35 40 45
Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

80
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro
1 5 10 15
Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp Met Asp
20 25 30
Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val
35 40 45
Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

111 amino acids

amino acid

linear

peptide

81
Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Val Pro
1 5 10 15
Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp Met Asp
20 25 30
Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala Phe Val
35 40 45
Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

113 amino acids

amino acid

linear

peptide

82
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp
20 25 30
Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

83
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp
20 25 30
Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

84
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp
20 25 30
Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

85
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

86
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

87
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

88
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

89
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

90
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

91
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

92
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

93
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

94
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

95
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

96
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln
100 105 110
Gln

339 base pairs

nucleic acid

double

linear

DNA (genomic)

97
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

98
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

99
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTTCCCTTGA GCACGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

100
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

101
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

102
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTTCCCTTGA GCACGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

103
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGGT TCCACCTGCA 60
CCTTTGCTGG ACAGTAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

104
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGGT TCCACCTGCA 60
CCTTTGCTGG ACAGTAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

105
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGGT TCCACCTGCA 60
CCTTTGCTGG ACAGTAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTTCCCTTGA GCACGCGCAG GAACAACAG 339

333 base pairs

nucleic acid

double

linear

DNA (genomic)

106
AACTGCTCTA TAATGATCGA TGAAATTATA CATCACTTAA AGAGACCACC TGCACCTTTG 60
CTGGACCCGA ACAACCTCAA TGCTGAAGAC GTCGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

107
AACTGCTCTA TAATGATCGA TGAAATTATA CATCACTTAA AGAGACCACC TAACCCTTTG 60
CTGGACCCGA ACAACCTCAA TTCTGAAGAC ATGGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

108
AACTGCTCTA TAATGATCGA TGAAATTATA CATCACTTAA AGGTTCCACC TGCACCTTTG 60
CTGGACAGTA ACAACCTCAA TTCCGAAGAC ATGGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

109
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAACG AAACCTTCGA 120
CTTCCAAACC TGCTCGCATT CGTAAGGGCT GTCAAGAACT TAGAAAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

110
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAACG AAACCTTCGA 120
CTTCCAAACC TGGAGAGCTT CGTAAGGGCT GTCAAGAACT TAGAAAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

111
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAACG AAACCTTCGA 120
ACTCCAAACC TGCTCGCATT CGTAAGGGCT GTCAAGCACT TAGAAAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

112
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGGTATT 180
GAGGCAATTC TTCGTAATCT CCAACCATGT CTGCCCTCTG CCACGGCCGC ACCCTCTCGA 240
CATCCAATCA TCATCAAGGC AGGTGACTGG CAAGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

113
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGGTATT 180
GAGGCAATTC TTCGTAATCT CGTACCATGT CTGCCCTCTG CCACGGCCGC ACCCTCTCGA 240
CATCCAATCA CCATCAAGGC AGGTGACTGG CAAGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

114
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GGGAAAAACT GACGTTCTAT 300
CTGGTTACCC TTGAGCAAGC GCAGGAACAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

115
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GGGAAAAACT GACGTTCTAT 300
CTGGTTTCCC TTGAGCACGC GCAGGAACAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

116
AACTGCTCTA TAATGATCGA TGAAATTATA CATCACTTAA AGAGACCACC TGCACCTTTG 60
CTGGACCCGA ACAACCTCAA TGCTGAAGAC GTCGATATCC TGATGGAACG AAACCTTCGA 120
CTTCCAAACC TGGAGAGCTT CGTAAGGGCT GTCAAGAACT TAGAAAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

117
AACTGCTCTA TAATGATCGA TGAAATTATA CATCACTTAA AGAGACCACC TAACCCTTTG 60
CTGGACCCGA ACAACCTCAA TTCTGAAGAC ATGGATATCC TGATGGAACG AAACCTTCGA 120
ACTCCAAACC TGCTCGCATT CGTAAGGGCT GTCAAGCACT TAGAAAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

118
AACTGCTCTA TAATGATCGA TGAAATTATA CATCACTTAA AGGTTCCACC TGCACCTTTG 60
CTGGACAGTA ACAACCTCAA TTCCGAAGAC ATGGATATCC TGATGGAACG AAACCTTCGA 120
CTTCCAAACC TGCTCGCATT CGTAAGGGCT GTCAAGAACT TAGAAAATGC ATCAGCAATT 180
GAGAGCATTC TTAAAAATCT CCTGCCATGT CTGCCCCTGG CCACGGCCGC ACCCACGCGA 240
CATCCAATCC ATATCAAGGA CGGTGACTGG AATGAATTCC GTCGTAAACT GACCTTCTAT 300
CTGAAAACCT TGGAGAACGC GCAGGCTCAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

119
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGGTATT 180
GAGGCAATTC TTCGTAATCT CCAACCATGT CTGCCCTCTG CCACGGCCGC ACCCTCTCGA 240
CATCCAATCA TCATCAAGGC AGGTGACTGG CAAGAATTCC GGGAAAAACT GACGTTCTAT 300
CTGGTTACCC TTGAGCAAGC GCAGGAACAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

120
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGGTATT 180
GAGGCAATTC TTCGTAATCT CGTACCATGT CTGCCCTCTG CCACGGCCGC ACCCTCTCGA 240
CATCCAATCA CCATCAAGGC AGGTGACTGG CAAGAATTCC GGGAAAAACT GACGTTCTAT 300
CTGGTTACCC TTGAGCAAGC GCAGGAACAA CAG 333

333 base pairs

nucleic acid

double

linear

DNA (genomic)

121
AACTGCTCTA ACATGATCGA TGAAATCATC ACCCACCTGA AGCAGCCACC GCTGCCGCTG 60
CTGGACTTCA ACAACCTCAA TGGTGAAGAC CAAGATATCC TGATGGAAAA TAACCTTCGT 120
CGTCCAAACC TCGAGGCATT CAACCGTGCT GTCAAGTCTC TGCAGAATGC ATCAGGTATT 180
GAGGCAATTC TTCGTAATCT CGTACCATGT CTGCCCTCTG CCACGGCCGC ACCCTCTCGA 240
CATCCAATCA CCATCAAGGC AGGTGACTGG CAAGAATTCC GGGAAAAACT GACGTTCTAT 300
CTGGTTTCCC TTGAGCACGC GCAGGAACAA CAG 333

339 base pairs

nucleic acid

double

linear

DNA (genomic)

122
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC 240
ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG AATTCCGTCG TAAACTGACC 300
TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

123
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC 240
ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG AATTCCGTCG TAAACTGACC 300
TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

124
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGGT TCCACCTGCA 60
CCTTTGCTGG ACAGTAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC 240
ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG AATTCCGTCG TAAACTGACC 300
TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

125
ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA GCCACCGCTG 60
CCGCTGCTGG ACTTCAACAA CCTCAATGGT GAAGACCAAG ATATCCTGAT GGAAAATAAC 120
CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA AGTCTCTGCA GAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

126
ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA GCCACCGCTG 60
CCGCTGCTGG ACTTCAACAA CCTCAATGGT GAAGACCAAG ATATCCTGAT GGAAAATAAC 120
CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA AGTCTCTGCA GAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

127
ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA GCCACCGCTG 60
CCGCTGCTGG ACTTCAACAA CCTCAATGGT GAAGACCAAG ATATCCTGAT GGAAAATAAC 120
CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA AGTCTCTGCA GAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCGTA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCACCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTTCCCTTGA GCACGCGCAG GAACAACAG 339

134 amino acids

amino acid

linear

peptide

128
Met Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn
1 5 10 15
Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro Pro
20 25 30
Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile
35 40 45
Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg
50 55 60
Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys
65 70 75 80
Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His
85 90 95
Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu
100 105 110
Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln Thr Thr
115 120 125
Leu Ser Leu Ala Ile Phe
130

133 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- may or may not precede
the amino acid in position 1”

Modified-site

/note= “Xaa at position 18 is Asn
or Ile”

Modified-site

/note= “Xaa at position 19 is Met,
Ala, or Ile”

Modified-site

/note= “Xaa at position 20 is Ile,
Pro, or Leu”

Modified-site

/note= “Xaa at position 23 is Ile,
Ala, or Leu”

Modified-site

/note= “Xaa at position 25 is Thr
or His”

Modified-site

/note= “Xaa at position 29 is Gln,
Arg, Val, or Leu”

Modified-site

/note= “Xaa at position 32 is Leu,
Ala, Asn, or Arg”

Modified-site

/note= “Xaa at position 34 is Leu
or Ser”

Modified-site

/note= “Xaa at position 37 is Phe,
Pro, or Ser”

Modified-site

; 38

/note= “Xaa at position 38 is Asn
or Ala”

Modified-site

/note= “Xaa at position 42 is Gly,
Ala, Ser, Asp, or Asn”

Modified-site

/note= “Xaa at position 45 is Gln,
Val, or Met”

Modified-site

/note= “Xaa at position 46 is Asp
or Ser”

Modified-site

/note= “Xaa at position 49 is Met,
Ile, Leu, or Asp”

Modified-site

/note= “Xaa at position 50 is Glu
or Asp”

Modified-site

/note= “Xaa at position 51 is Asn,
Arg, or Ser”

Modified-site

/note= “Xaa at position 55 is Arg,
Leu, or Thr”

Modified-site

/note= “Xaa at position 56 is Pro
or Ser”

Modified-site

/note= “Xaa at position 59 is Glu
or Leu”

Modified-site

/note= “Xaa at position 60 is Ala
or Ser”

Modified-site

/note= “Xaa at position 62 is Asn
Val, or Pro”

Modified-site

/note= “Xaa at position 63 is Arg
or His”

Modified-site

/note= “Xaa at position 65 is Val
or Ser”

Modified-site

/note= “Xaa at position 67 is Ser,
Asn, His, or Gly”

Modified-site

/note= “Xaa at position 69 is Gln
or Glu”

Modified-site

/note= “Xaa at position 73 is Ala
or Gly”

Modified-site

/note= “Xaa at position 76 is Ser,
Ala, or Pro”

Modified-site

/note= “Xaa at position 79 is Lys,
Arg, or Ser”

Modified-site

/note= “Xaa at position 82 is Leu,
Glu, Val, or Trp”

Modified-site

/note= “Xaa at position 85 is Leu
or Val”

Modified-site

/note= “Xaa at position 87 is Leu,
Ser, or Trp”

Modified-site

/note= “Xaa at position 88 is Ala
or Trp”

Modified-site

/note= “Xaa at position 91 is Ala
or Pro”

Modified-site

/note= “Xaa at position 93 is Pro
or Ser”

Modified-site

/note= “Xaa at position 95 is His
or Thr”

Modified-site

/note= “Xaa at position 98 is His,
Ile, or Thr”

Modified-site

100

/note= “Xaa at position 100 is Lys
or Arg”

Modified-site

101

/note= “Xaa at position 101 is Asp,
Ala, or Met”

Modified-site

105

/note= “Xaa at position 105 is Asn
or Gln”

Modified-site

109

/note= “Xaa at position 109 is Arg,
Glu, or Leu”

Modified-site

112

/note= “Xaa at position 112 is Thr
or Gln”

Modified-site

116

/note= “Xaa at position 116 is Lys,
Val, Trp, or Ser”

Modified-site

117

/note= “Xaa at position 117 is Thr
or Ser”

Modified-site

120

/note= “Xaa at position 120 is Asn,
Gln, or His”

Modified-site

123

/note= “Xaa at position 123 is Ala
or Glu”

129
Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn Cys
1 5 10 15
Ser Xaa Xaa Xaa Asp Glu Xaa Ile Xaa His Leu Lys Xaa Pro Pro Xaa
20 25 30
Pro Xaa Leu Asp Xaa Xaa Asn Leu Asn Xaa Glu Asp Xaa Xaa Ile Leu
35 40 45
Xaa Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa Xaa Ala
50 55 60
Xaa Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa Ile Leu Xaa Asn
65 70 75 80
Leu Xaa Pro Cys Xaa Pro Xaa Xaa Thr Ala Xaa Pro Xaa Arg Xaa Pro
85 90 95
Ile Xaa Ile Xaa Xaa Gly Asp Trp Xaa Glu Phe Arg Xaa Lys Leu Xaa
100 105 110
Phe Tyr Leu Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln Thr Thr Leu
115 120 125
Ser Leu Ala Ile Phe
130

111 amino acids

amino acid

linear

peptide

Modified-site

/note= “Met- or Met-Ala may or may
not precede the amino acid in position 1”

Modified-site

/note= “Xaa at position 4 is Asn or
Ile”

Modified-site

/note= “Xaa at position 5 is Met,
Ala, or Ile”

Modified-site

/note= “Xaa at position 6 is Ile,
Pro, or Leu”

Modified-site

/note= “Xaa at position 9 is Ile,
Ala, or Leu”

Modified-site

/note= “Xaa at position 11 is Thr
or His”

Modified-site

/note= “Xaa at position 15 is Gln,
Arg, Val, or Leu”

Modified-site

/note= “Xaa at position 18 is Leu,
Ala, Asn, or Arg”

Modified-site

/note= “Xaa at position 20 is Leu
or Ser”

Modified-site

/note= “Xaa at position 23 is Phe,
Pro, or Ser”

Modified-site

/note= “Xaa at position 24 is Asn
or Ala”

Modified-site

/note= “Xaa at position 28 is Gly,
Ala, Ser, Asp, or Asn”

Modified-site

/note= “Xaa at position 31 is Gln,
Val, or Met”

Modified-site

/note= “Xaa at position 32 is Asp
or Ser”

Modified-site

/note= “Xaa at position 35 is Met,
Ile, Leu, or Asp”

Modified-site

/note= “Xaa at position 36 is Glu
or Asp”

Modified-site

/note= “Xaa at position 37 is Asn,
Arg, or Ser”

Modified-site

/note= “Xaa at position 41 is Arg,
Leu, or Thr”

Modified-site

/note= “Xaa at position 42 is Pro
or Ser”

Modified-site

/note= “Xaa at position 45 is Glu
or Leu”

Modified-site

/note= “Xaa at position 46 is Ala
or Ser”

Modified-site

/note= “Xaa at position 48 is Asn,
Val, or Pro”

Modified-site

/note= “Xaa at position 49 is Arg
or His”

Modified-site

/note= “Xaa at position 51 is Val
or Ser”

Modified-site

/note= “Xaa at position 53 is Ser,
Asn, His, or Gly”

Modified-site

/note= “Xaa at position 55 is Gln
or Glu”

Modified-site

/note= “Xaa at position 59 is Ala
or Gly”

Modified-site

/note= “Xaa at position 62 is Ser,
Ala, or Pro”

Modified-site

/note= “Xaa at position 65 is Lys,
Arg, or Ser”

Modified-site

/note= “Xaa at position 67 is Leu,
Glu, or Val”

Modified-site

/note= “Xaa at position 68 is Leu,
Glu, Val, or Trp”

Modified-site

/note= “Xaa at position 71 is Leu
or Val”

Modified-site

/note= “Xaa at position 73 is Leu,
Ser, or Trp”

Modified-site

/note= “Xaa at position 74 is Ala
or Trp”

Modified-site

/note= “Xaa at position 77 is Ala
or Pro”

Modified-site

/note= “Xaa at position 79 is Pro
or Ser”

Modified-site

/note= “Xaa at position 81 is His
or Thr”

Modified-site

/note= “Xaa at position 84 is His,
Ile, or Thr”

Modified-site

/note= “Xaa at position 86 is Lys
or Arg”

Modified-site

/note= “Xaa at position 87 is Asp,
Ala, or Met”

Modified-site

/note= “Xaa at position 91 is Asn
or Glu”

Modified-site

/note= “Xaa at position 95 is Arg,
Glu, or Leu”

Modified-site

/note= “Xaa at position 98 is Thr
or Gln”

Modified-site

102

/note= “Xaa at position 102 is Lys,
Val, Trp, or Ser”

Modified-site

103

/note= “Xaa at position 103 is Thr
or Ser”

Modified-site

106

/note= “Xaa at position 106 is Asn,
Gln, or His”

Modified-site

109

/note= “Xaa at position 109 is Ala
or Glu”

130
Asn Cys Ser Xaa Xaa Xaa Asp Glu Xaa Ile Xaa His Leu Lys Xaa Pro
1 5 10 15
Pro Xaa Pro Xaa Leu Asp Xaa Xaa Asn Leu Asn Xaa Glu Asp Xaa Xaa
20 25 30
Ile Leu Xaa Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa
35 40 45
Xaa Ala Xaa Lys Xaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa Ile Leu
50 55 60
Xaa Asn Xaa Xaa Pro Cys Xaa Pro Xaa Xaa Thr Ala Xaa Pro Xaa Arg
65 70 75 80
Xaa Pro Ile Xaa Ile Xaa Xaa Gly Asp Trp Xaa Glu Phe Arg Xaa Lys
85 90 95
Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln
100 105 110

58 base pairs

nucleic acid

single

linear

DNA (synthetic)

131
CTAGCCACGG CCGCACCCAC GCGACATCCA ATCCATATCA AGGACGGTGA CTGGAATG 58

58 base pairs

nucleic acid

single

linear

DNA (synthetic)

132
TTAACATTCC AGTCACCGTC CTTGATATGG ATTGGATGTC GCGTGGGTGC GGCCGTGG 58

29 base pairs

nucleic acid

double

linear

DNA (genomic)

133
AAGGAGATAT ATCCATGAAC TGCTCTAAC 29

5 amino acids

amino acid

linear

peptide

134
Met Asn Cys Ser Asn
1 5

120 amino acids

amino acid

linear

peptide

135
Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln
1 5 10 15
Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln
20 25 30
Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe
35 40 45
Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile
50 55 60
Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr
65 70 75 80
Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg
85 90 95
Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110
Thr Thr Leu Arg Leu Ala Ile Phe
115 120

71 base pairs

nucleic acid

single

linear

DNA (synthetic)

136
AATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAGA 60
CCACTCTGTC G 71

71 base pairs

nucleic acid

single

linear

DNA (synthetic)

137
CTAGCGACAG AGTGGTCTGT TGAGCCTGCG CGTTCTCCAA GGTTTTCAGA TAGAAGGTCA 60
GTTTACGACG G 71

112 amino acids

amino acid

linear

peptide

138
Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln
1 5 10 15
Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln
20 25 30
Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe
35 40 45
Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile
50 55 60
Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr
65 70 75 80
Arg His Pro Ile His Ile Lys Ala Gly Asp Trp Asn Glu Phe Arg Arg
85 90 95
Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

75 amino acids

amino acid

linear

peptide

139
Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln
1 5 10 15
Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln
20 25 30
Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe
35 40 45
Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile
50 55 60
Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala
65 70 75

58 base pairs

nucleic acid

single

linear

DNA (synthetic)

140
CTAGCCACGG CCGCACCCAC GCGACATCCA ATCCATATCA AGGCTGGTGA CTGGAATG 58

58 base pairs

nucleic acid

single

linear

DNA (synthetic)

141
AATTCATTCC AGTCACCAGC CTTGATATGG ATTGGATGTC GCGTGGGTGC GGCCGTGG 58

64 base pairs

nucleic acid

single

linear

DNA (synthetic)

142
AATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAGT 60
AATA 64

64 base pairs

nucleic acid

single

linear

DNA (synthetic)

143
AGCTTATTAC TGTTGAGCCT GCGCGTTCTC CAAGGTTTTC AGATAGAAGG TCAGTTTACG 60
ACGG 64

408 base pairs

nucleic acid

double

linear

DNA (genomic)

144
ATGGCTCCAA TGACTCAGAC TACTTCTCTT AAGACTTCTT GGGTTAACTG CTCTAACATG 60
ATCGATGAAA TTATAACACA CTTAAAGCAG CCACCTTTGC CTTTGCTGGA CTTCAACAAC 120
CTCAATGGGG AAGACCAAGA CATTCTGATG GAAAATAACC TTCGAAGGCC AAACCTGGAG 180
GCATTCAACA GGGCTGTCAA GAGTTTACAG AATGCATCAG CAATTGAGAG CATTCTTAAA 240
AATCTCCTGC CATGTCTGCC CCTGGCCACG GCCGCACCCA CGCGACATCC AATCCATATC 300
AAGGACGGTG ACTGGAATGA ATTCCGTCGT AAACTGACCT TCTATCTGAA AACCTTGGAG 360
AACGCGCAGG CTCAACAGAC CACTCTGTCG CTAGCGATCT TTTAATAA 408

157 base pairs

nucleic acid

double

linear

DNA (genomic)

CDS

1..156

145
ATC GAT GAA ATC ATC ACC CAC CTG AAG CAG CCA CCG CTG CCG CTG CTG 48
Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu
1 5 10 15
GAC TTC AAC AAC CTC AAT GGT GAA GAC CAA GAT ATC CTG ATG GAA AAT 96
Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn
20 25 30
AAC CTT CGT CGT CCA AAC CTC GAG GCA TTC AAC CGT GCT GTC AAG TCT 144
Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser
35 40 45
CTG CAG AAT GCA T 157
Leu Gln Asn Ala
50

52 amino acids

amino acid

linear

protein

146
Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu
1 5 10 15
Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn
20 25 30
Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser
35 40 45
Leu Gln Asn Ala
50

414 base pairs

nucleic acid

double

linear

DNA (genomic)

147
CCATGGCTCC AATGACTCAG ACTACTTCTC TTAAGACTTC TTGGGTTAAC TGCTCTAACA 60
TGATCGATGA AATTATAACA CACTTAAAGC AGCCACCTTT GCCTTTGCTG GACTTCAACA 120
ACCTCAATGG GGAAGACCAA GACATTCTGA TGGAAAATAA CCTTCGAAGG CCAAACCTGG 180
AGGCATTCAA CAGGGCTGTC AAGAGTTTAC AGAATGCATC AGCAATTGAG AGCATTCTTA 240
AAAATCTCCT GCCATGTCTG CCCCTGGCCA CGGCCGCACC CACGCGACAT CCAATCCATA 300
TCAAGGACGG TGACTGGAAT GAATTCCGTC GTAAACTGAC CTTCTATCTG AAAACCTTGG 360
AGAACGCGCA GGCTCAACAG ACCACTCTGT CGCTAGCGAT CTTTTAATAA GCTT 414

414 base pairs

nucleic acid

double

linear

DNA (genomic)

148
AAGCTTATTA AAAGATCGCT AGCGACAGAG TGGTCTGTTG AGCCTGCGCG TTCTCCAAGG 60
TTTTCAGATA GAAGGTCAGT TTACGACGGA ATTCATTCCA GTCACCGTCC TTGATATGGA 120
TTGGATGTCG CGTGGGTGCG GCCGTGGCCA GGGGCAGACA TGGCAGGAGA TTTTTAAGAA 180
TGCTCTCAAT TGCTGATGCA TTCTGTAAAC TCTTGACAGC CCTGTTGAAT GCCTCCAGGT 240
TTGGCCTTCG AAGGTTATTT TCCATCAGAA TGTCTTGGTC TTCCCCATTG AGGTTGTTGA 300
AGTCCAGCAA AGGCAAAGGT GGCTGCTTTA AGTGTGTTAT AATTTCATCG ATCATGTTAG 360
AGCAGTTAAC CCAAGAAGTC TTAAGAGAAG TAGTCTGAGT CATTGGAGCC ATGG 414

81 base pairs

nucleic acid

double

linear

DNA (genomic)

149
ATGATGATTA CTCTGCGCAA ACTTCCTCTG GCGGTTGCCG TCGCAGCGGG CGTAATGTCT 60
GCTCAGGCCA TGGCTAACTG C 81

81 base pairs

nucleic acid

double

linear

DNA (genomic)

150
GCAGTTAGCC ATGGCCTGAG CAGACATTAC GCCCGCTGCG ACGGCAACCG CCAGAGGAAG 60
TTTGCGCAGA GTAATCATCA T 81

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

151
CATGGCTAAC TGCTCTAACA TGAT 24

22 base pairs

nucleic acid

single

linear

DNA (synthetic)

152
CGATCATGTT AGAGCAGTTA GC 22

12 base pairs

nucleic acid

double

linear

DNA (genomic)

153
ATGGCTAACT GC 12

4 amino acids

amino acid

linear

peptide

154
Met Ala Asn Cys
1

34 base pairs

nucleic acid

single

linear

DNA (synthetic)

155
GCCGATACCG CGGCATACTC CCACCATTCA GAGA 34

33 base pairs

nucleic acid

single

linear

DNA (synthetic)

156
GCCGATAAGA TCTAAAACGG GTATGGAGAA ACA 33

30 base pairs

nucleic acid

single

linear

DNA (synthetic)

157
ATAGTCTTCC CCAGATATCT AACGCTTGAG 30

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

158
CAATACCTGA TGCGTTTTCT AAGT 24

33 base pairs

nucleic acid

single

linear

DNA (synthetic)

159
GGTTTCGTTC CATCAGAATG TCCATGTCTT CAG 33

339 base pairs

nucleic acid

double

linear

DNA (genomic)

160
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAACGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

338 base pairs

nucleic acid

double

linear

DNA (genomic)

161
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACATGG ACATTTGATG GAACGAAACC 120
TTCGAACTCC AAACCTGCTC GCATTCGTAA GGGCTGTCAA GCACTTAGAA AACGCATCAG 180
GTATTGAGGC AATTCTTCGT AATCTCCAAC CATGTCTGCC CTCTGCCACG GCCGCACCCT 240
CTCGACATCC AATCATCATC AAGGCAGGTG ACTGGCAAGA ATTCCGGGAA AAACTGACGT 300
TCTATCTGGT TACCCTTGAG CAAGCGCAGG AACAACAG 338

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

162
CATGGCTAAC TGCTCTAACA TGATCGATGA AATTATAACA 40

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

163
CTTTAAGTGT GTTATAATTT CATCGATCAT GTTAGAGCAG TTAGC 45

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

164
CACTTAAAGC AGCCACCTTT GCCTTTGCTG GACTTC 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

165
GAGGTTGTTG AAGTCCAGCA AAGGCAAAGG TGGCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

166
AACAACCTCA ATGACGAAGA CATGTCT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

167
AGACATGTCT TCGTCATT 18

16 base pairs

nucleic acid

single

linear

DNA (synthetic)

168
TGAACCATAT GTCAGG 16

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

169
AATTCCTGAC ATATGGTTCA TGCA 24

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

170
CATGGCAAAC TGCTCTATAG CTATCGATGA AATTATACAT 40

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

171
CTTTAAGTGA TGTATAATTT CATCGATAGC TATAGAGCAG TTTGC 45

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

172
CATGGCAAAC TGCTCTATAA TCATCGATGA AATTATACAT 40

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

173
CTTTAAGTGA TGTATAATTT CATCGATGAT TATAGAGCAG TTTGC 45

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

174
ATCCTGGACG AACGAAACCT TCGAACTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

175
AGTTCGAAGG TTTCGTTCGT CCAGGAT 27

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

176
ATCCTGATCG AACGAAACCT TCGAACTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

177
AGTTCGAAGG TTTCGTTCGA TCAGGAT 27

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

178
ATCCTGCTGG AACGAAACCT TCGAACTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

179
AGTTCGAAGG TTTCGTTCCA GCAGGAT 27

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

180
AACAACCTCA ATTCTGAAGA CGTTGAT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

181
ATCAACGTCT TCAGAATT 18

51 base pairs

nucleic acid

single

linear

DNA (synthetic)

182
CGCGCCATGG CTAACTGCTC TATAATGATC GATGAAGCAA TACATCACTT A 51

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

183
CGCGTCGATA AGCTTATT 18

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

184
GGAGATATAT CCATGGCT 18

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

185
TCGGTCCATC AGAATAGACA TGTCTTCAGC ATTGAGGTTG TT 42

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

186
TCGGTCCATC AGAATAGAAA CGTCTTCAGC ATTGAGGTTG TT 42

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

187
TCGGTCCATC AGAATAGACA TGTCTTCGTC ATTGAGGTTG TT 42

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

188
TCGGTCCATC AGAATAGAAA CGTCTTCGTC ATTGAGGTTG TT 42

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

189
TCGGTCCATC AGAATAGACA TGTCTTCAGA ATTGAGGTTG TT 42

42 base pairs

nucleic acid

single

linear

DNA (synthetic)

190
TCGGTCCATC AGAATAGAAA CGTCTTCAGA ATTGAGGTTG TT 42

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

191
CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCA TCATCCGT 48

26 base pairs

nucleic acid

single

linear

DNA (synthetic)

192
AATTCTTGCC AGTCACCTGC ACGGAT 26

16 base pairs

nucleic acid

single

linear

DNA (synthetic)

193
ATGGGTGACT GGCAAG 16

26 base pairs

nucleic acid

single

linear

DNA (synthetic)

194
AATTCTTGCC AGTCACCCAT ACGGAT 26

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

195
CATGGCTAAC TGCTCTATTA TGATCGATGA AGCAATACAT 40

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

196
CTTTAAGTGA TGTATTGCTT CATCGATCAT AATAGAGCAG TTAGC 45

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

197
CACTTAAAGG TACCACCTCG CCCTTCCCTG GACCCG 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

198
GAGGTTGTTC GGGTCCAGGG AAGGGCGAGG TGGTAC 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

199
CACTTAAAGA GACCACCTGC ACCTTCCCTG GACCCG 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

200
GAGGTTGTTC GGGTCCAGGG AAGGTGCAGG TGGTCT 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

201
AACAACCTCA ATGACGAAGA CATGGAT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

202
ATCCATGTCT TCGTCATT 18

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

203
AACAACCTCA ATGACGAAGA CGTCGAT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

204
ATCGACGTCT TCGTCATT 18

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

205
AACAACCTCA ATGACGAAGA CATGTCT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

206
AGACATGTCT TCGTCATT 18

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

207
AACAACCTCA ATGACGAAGA CGTCTCT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

208
AGAGACGTCT TCGTCATT 18

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

209
ATCCTGATGG ACCGAAACCT TCGACTTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

210
AAGTCGAAGG TTTCGGTCCA TCAGGAT 27

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

211
ATCCTGATGG ACCGAAACCT TCGACTTAGC AACCTG 36

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

212
CCTTACGAAG CTCTCCAGGT TGCT 24

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

213
CGTAATCTCT GGCCATGT 18

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

214
CCAGAGATTA CGAAGAAT 18

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

215
AATTCCGGGA AAAACTGCAA TTCTATCTGT GG 32

37 base pairs

nucleic acid

single

linear

DNA (synthetic)

216
CTCAAGGGTC CACAGATAGA ATTGCAGTTT TTCCCGG 37

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

217
AATTCCGGGA AAAACTGCAA TTCTATCTGG TT 32

37 base pairs

nucleic acid

single

linear

DNA (synthetic)

218
CTCAAGGGTA ACCAGATAGA ATTGCAGTTT TTCCCGG 37

23 base pairs

nucleic acid

single

linear

DNA (synthetic)

219
AATTCCGGGA AAAACTGACG TTC 23

28 base pairs

nucleic acid

single

linear

DNA (synthetic)

220
AACCAGATAG AACGTCAGTT TTTCCCGG 28

23 base pairs

nucleic acid

single

linear

DNA (synthetic)

221
TATCTGGTTA CCCTTGAGTA ATA 23

19 base pairs

nucleic acid

single

linear

DNA (synthetic)

222
AGCTTATTAC TTCAAGGGT 19

23 base pairs

nucleic acid

single

linear

DNA (synthetic)

223
AATTCCGGGA AAAACTGCAA TTC 23

28 base pairs

nucleic acid

single

linear

DNA (synthetic)

224
AACCAGATAG AATTGCAGTT TTTCCCGG 28

61 base pairs

nucleic acid

single

linear

DNA (synthetic)

225
CGATCATTAT AGAGCAGTTA GCCTTGTCAT CGTCGTCCTT GTAATCAGTT TCTGGATATG 60
C 61

63 base pairs

nucleic acid

single

linear

DNA (synthetic)

226
CATGGCATAT CCAGAAACTG ATTACAAGGA CGACGATGAC AAGGCTAACT GCTCTATAAT 60
GAT 63

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

227
AATTCCGGCT TAAACTGCAA TTCTATCTGT CT 32

37 base pairs

nucleic acid

single

linear

DNA (synthetic)

228
CTCAAGGGTA GACAGATAGA ATTGCAGTTT AAGCCGG 37

32 base pairs

nucleic acid

single

linear

DNA (synthetic)

229
TCTCTTGAGC AAGCGCAGGA ACAACAGTAA TA 32

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

230
CATGGCAAAC TGCTCTATAA TACTCGATGA AGCAATACAT 40

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

231
CTTTAAGTGA TGTATTGCTT CATCGAGTAT TATAGAGCAG TTTGC 45

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

232
CATGGCAAAC TGCTCTATAA TGCCAGATGA AGCAATACAT 40

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

233
CTTTAAGTGA TGTATTGCTT CATCTGGCAT TATAGAGCAG TTTGC 45

41 base pairs

nucleic acid

single

linear

DNA (synthetic)

234
CATGGCAAAC TGCTCTATAA TGATCGATGA AACTGATACA T 41

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

235
CTTTAAGTGA TGTATCAGTT CATCGATCAT TATAGAGCAG TTTGC 45

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

236
CACTTAAAGA TACCACCTAA CCCTAGCCTG GACAGT 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

237
GAGGTTAGCA CTGTCCAGGC TAGGGTTAGG TGGTAT 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

238
GCTAACCTCA ATTCCGAAGA CGTCTCT 27

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

239
AGAGACGTCT TCGGAATT 18

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

240
ATCCTGATGG ACTCCAACCT TCGAACTCCA AACCTG 36

27 base pairs

nucleic acid

single

linear

DNA (synthetic)

241
AGTTCGAAGG TTGGAGTCCA TCAGGAT 27

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

242
GTTCCCTATT GGACGGCCCC TCCCTCTCGA ACACCAATCA CGATCAAG 48

48 base pairs

nucleic acid

single

linear

DNA (synthetic)

243
CGTGATTGGT GTTCGAGAGG GAGGGGCCGT CCAATAGGGA ACACATGG 48

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

244
CTCGCATTCC CACATGCTTC TAAG 24

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

245
CTCGCATTCC CACATGCTGT CAAG 24

24 base pairs

nucleic acid

single

linear

DNA (synthetic)

246
ATGTGGGAAT GCGAGCAGGT TTGG 24

20 base pairs

nucleic acid

single

linear

DNA (synthetic)

247
TTTTCTAATT GCTTAGAAGC 20

15 base pairs

nucleic acid

single

linear

DNA (synthetic)

248
CAATTAGAAA ATGCA 15

20 base pairs

nucleic acid

single

linear

DNA (synthetic)

249
TTTTCTAATT GCTTGACAGC 20

21 base pairs

nucleic acid

single

linear

DNA (synthetic)

250
TCAGGTATTG AGCCAATTCT T 21

19 base pairs

nucleic acid

single

linear

DNA (synthetic)

251
TGGCTCAATA CCTGATGCA 19

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

252
TCTAATCTCC AACCATGT 18

18 base pairs

nucleic acid

single

linear

DNA (synthetic)

253
TTGGAGATTA GAAAGAAT 18

37 base pairs

nucleic acid

single

linear

DNA (synthetic)

254
CTCAAGAGAA GACAGATAGA ATTGCAGTTT AAGCCGG 37

40 base pairs

nucleic acid

single

linear

DNA (synthetic)

255
CATGGCTAAC TGCTCTATAA TGATCGATGA AATTATACAT 40

43 base pairs

nucleic acid

single

linear

DNA (synthetic)

256
AATTCCGGCT TAAACTGCAA TTCTATCTGT CTACCCTTTA ATA 43

43 base pairs

nucleic acid

single

linear

DNA (synthetic)

257
AGCTTATTAA AGGGTAGACA GATAGAATTG CAGTTTAAGC CGG 43

43 base pairs

nucleic acid

single

linear

DNA (synthetic)

258
AATTCCGGCT TAAACTGCAA TTCTATCTGT CTACCCTTTA ATA 43

113 amino acids

amino acid

linear

peptide

259
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

260
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

261
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Trp Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

262
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Trp Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

263
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg Arg Pro Ile Ile Ile Arg Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Trp Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

264
Met Ala Asn Cys Ser Ile Ala Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

265
Met Ala Asn Cys Ser Ile Ile Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

266
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

267
Met Ala Asn Cys Ser Ile Ala Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

268
Met Ala Asn Cys Ser Ile Ile Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

269
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Ile Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

270
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Leu Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

271
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Asp Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

272
Met Ala Asn Cys Ser Ile Ala Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Ile Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

273
Met Ala Asn Cys Ser Ile Ile Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Ile Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

274
Met Ala Asn Cys Ser Ile Ala Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Leu Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

275
Met Ala Asn Cys Ser Ile Ile Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Leu Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

276
Met Ala Asn Cys Ser Ile Ala Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Asp Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

277
Met Ala Asn Cys Ser Ile Ile Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Val Asp Ile Leu Asp Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

278
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

279
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

280
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

281
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asn Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

282
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

283
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

284
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

285
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

286
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

107 amino acids

amino acid

linear

peptide

287
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu
100 105

107 amino acids

amino acid

linear

peptide

288
Met Asp Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu
100 105

113 amino acids

amino acid

linear

peptide

289
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

107 amino acids

amino acid

linear

peptide

290
Met Asp Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu
100 105

113 amino acids

amino acid

linear

peptide

291
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

292
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Val Pro Pro Arg Pro Ser Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

293
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Trp Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

294
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

295
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

296
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Asp Arg Asn Leu Arg Leu Ser Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

297
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

298
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Ser Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

299
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

300
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Ser Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

301
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

302
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Val Pro Pro Arg Pro Ser Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

303
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Arg Met Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

304
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Trp Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Arg Met Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

305
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

306
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Val Pro Pro Arg Pro Ser Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

107 amino acids

amino acid

linear

peptide

307
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Val Thr Leu Glu
100 105

113 amino acids

amino acid

linear

peptide

308
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

309
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

310
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

311
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Ser Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Val Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Ser Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

312
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Ser Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

313
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Gln Phe Tyr Leu Trp Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

314
Met Asp Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp
20 25 30
Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

125 amino acids

amino acid

linear

peptide

315
Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp Asp Lys Asn Cys
1 5 10 15
Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro Pro Ala
20 25 30
Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu
35 40 45
Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala
50 55 60
Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn
65 70 75 80
Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro
85 90 95
Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr
100 105 110
Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln
115 120 125

125 amino acids

amino acid

linear

peptide

316
Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp Asp Lys Asn Cys
1 5 10 15
Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro Pro Asn
20 25 30
Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu
35 40 45
Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val Arg Ala
50 55 60
Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn
65 70 75 80
Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro
85 90 95
Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr
100 105 110
Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln
115 120 125

113 amino acids

amino acid

linear

peptide

317
Met Ala Asn Cys Ser Ile Met Pro Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

318
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Leu Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

319
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Pro
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg Thr Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

320
Met Ala Asn Cys Ser Ile Ile Leu Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Pro
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg Thr Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

321
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Asp Ser Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Pro His Ala Ser Lys Gln Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

322
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

106 amino acids

amino acid

linear

peptide

323
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Thr Leu
100 105

113 amino acids

amino acid

linear

peptide

324
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Ser Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

325
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Asp Ser Asn Leu Leu Thr Pro Asn Leu Leu Ala
35 40 45
Phe Pro His Ala Ser Lys Gln Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Ser Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

326
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Pro
50 55 60
Ile Leu Ser Asn Leu Gln Pro Cys Val Pro Tyr Trp Thr Ala Pro Pro
65 70 75 80
Ser Arg Thr Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

327
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp
20 25 30
Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Pro
50 55 60
Ile Leu Ser Asn Leu Gln Pro Cys Val Pro Tyr Trp Thr Ala Pro Pro
65 70 75 80
Ser Arg Thr Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

328
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Leu Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Pro
50 55 60
Ile Leu Ser Asn Leu Gln Pro Cys Val Pro Tyr Trp Thr Ala Pro Pro
65 70 75 80
Ser Arg Thr Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

329
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Leu Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Leu Lys Leu Gln Phe Tyr Leu Ser Ser Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

330
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Leu Ile His His Leu Lys
1 5 10 15
Ile Pro Pro Asn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser Glu Asp
20 25 30
Val Ser Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala
35 40 45
Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Pro
50 55 60
Ile Leu Ser Asn Leu Gln Pro Cys Val Pro Tyr Trp Thr Ala Pro Pro
65 70 75 80
Ser Arg Thr Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

113 amino acids

amino acid

linear

peptide

331
Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ala Ile His His Leu Lys
1 5 10 15
Arg Pro Pro Ala Pro Ser Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Asp Arg Asn Leu Arg Leu Ser Asn Leu Glu Ser
35 40 45
Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu Ala
50 55 60
Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro
65 70 75 80
Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg
85 90 95
Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln
100 105 110
Gln

339 base pairs

nucleic acid

double

linear

DNA (genomic)

332
ATGGCAAACT GCTCTATAGC TATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

333
ATGGCAAACT GCTCTATAAT CATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

334
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

335
ATGGCAAACT GCTCTATAGC TATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

336
ATGGCAAACT GCTCTATAAT GATCCATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

337
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT CGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

338
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGCT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

339
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGGA CGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

340
ATGGCAAACT GCTCTATAGC TATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGAT CGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

341
ATGGCAAACT GCTCTATAAT CATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGAT CGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

342
ATGGCAAACT GCTCTATAGC TATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGCT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

343
ATGGCAAACT GCTCTATAAT CATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGCT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

344
ATGGCAAACT GCTCTATAGC TATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGGA CGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

345
ATGGCAAACT GCTCTATAAT CATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCT GAAGACGTTG ATATCCTGGA CGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

346
ATGGCTAACT GCTCTATTAT GATCGATGAA GCAATACATC ACTTAAAGGT TCCACCTGCA 60
CCTTTGCTGG ACAGTAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

347
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGGT TCCACCTGCA 60
CCTTTGCTGG ACAGTAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

348
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTTT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

349
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATAAC GAAGACGTTT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

350
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTTT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

351
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTTT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

352
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACATGT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

353
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACATGT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

354
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

321 base pairs

nucleic acid

double

linear

DNA (genomic)

355
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA G 321

321 base pairs

nucleic acid

double

linear

DNA (genomic)

356
ATGGATAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA G 321

321 base pairs

nucleic acid

double

linear

DNA (genomic)

357
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA G 321

321 base pairs

nucleic acid

double

linear

DNA (genomic)

358
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA G 321

339 base pairs

nucleic acid

double

linear

DNA (genomic)

359
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTTT CTATCCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

360
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGGT ACCACCTCGC 60
CCTTCCCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

361
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCTGG CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

362
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTTT CTATTCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

363
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

364
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGACCGAAAC 120
CTTCGACTTA GCAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

365
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

366
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTCCCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

367
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

368
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTCCCTGG ACCCGAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

369
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

370
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGGT ACCACCTCGC 60
CCTTCCCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

371
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CCGTATGGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

372
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCTGG CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CCGTATGGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

373
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

374
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGGT ACCACCTCGC 60
CCTTCCCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

321 base pairs

nucleic acid

double

linear

DNA (genomic)

375
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGG TTACCCTTGA G 321

339 base pairs

nucleic acid

double

linear

DNA (genomic)

376
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

377
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCG ATTCTCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

378
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCA TGTCTCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

379
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGAC 60
CTTTCCCTGG ACCCGAACAA CCTCAATGAC GAAGACGTCT CTATCCTGAT GGACCGAAAC 120
CTTCGACTTA GCAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

380
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTCCCTGG ACCCGAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGACCGAAAC 120
CTTCGACTTA GCAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

381
ATGGCTAACT GCTCTATAAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTCCCTGG ACCCGAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGACCGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

382
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGCAA 300
TTCTATCTGT GGACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

383
ATGGATAACT GCTCTATTAT GATCGATGAA GCAATACATC ACTTAAAGAG ACCACCTGCA 60
CCTTTGCTGG ACCCGAACAA CCTCAATGCT GAAGACGTCG ATATCCTGAT GGAACGAAAC 120
CTTCGACTTC CAAACCTGGA GAGCTTCGTA AGGGCTGTCA AGAACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

384
ATGGCTAACT GCTCTATAAT GCCAGATGAA GCAATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

385
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

386
ATGGCTAACT GCTCTATTAT GATCGATGAA GCAATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

387
ATGGCTAACT GCTCTATAAT ACTCGATGAA GCAATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

388
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGACTCCAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCCCA CATGCTGTCA AGCAATTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

389
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGACTCCAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCCCA CATGCTTCTA AGCAATTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

390
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGCT TAAACTGCAA 300
TTCTATCTGT CTACCCTTGA GCAAGCGCAG GAACAACAG 339

318 base pairs

nucleic acid

double

linear

DNA (genomic)

391
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGCT TAAACTGCAA 300
TTCTATCTGT CTACCCTT 318

339 base pairs

nucleic acid

double

linear

DNA (genomic)

392
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGCT TAAACTGCAA 300
TTCTATCTGT CTTCTCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

393
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGC CAATTCTTTC TAATCTCCAA CCATGTGTTC CCTATTGGAC GGCCCCTCCC 240
TCTCGAACAC CAATCACGAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

394
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAG ACCACCTAAC 60
CCTTTGCTGG ACCCGAACAA CCTCAATTCC GAAGACATGG ATATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGC CAATTCTTTC TAATCTCCAA CCATGTGTTC CCTATTGGAC GGCCCCTCCC 240
TCTCGAACAC CAATCACGAT CAAGGCAGGT GACTGGCAAG AATTCCGGCT TAAACTGCAA 300
TTCTATCTGT CTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

395
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGC CAATTCTTTC TAATCTCCAA CCATGTGTTC CCTATTGGAC GGCCCCTCCC 240
TCTCGAACAC CAATCACGAT CAAGGCAGGT GACTGGCAAG AATTCCGGCT TAAACTGCAA 300
TTCTATCTGT CTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

396
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGG CAATTCTTCG TAATCTCCAA CCATGTCTGC CCTCTGCCAC GGCCGCACCC 240
TCTCGACATC CAATCATCAT CAAGGCAGGT GACTGGCAAG AATTCCGGCT TAAACTGCAA 300
TTCTATCTGT CTTCTCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

397
ATGGCTAACT GCTCTATAAT GATCGATGAA ATTATACATC ACTTAAAGAT ACCACCTAAC 60
CCTAGCCTGG ACAGTGCTAA CCTCAATTCC GAAGACGTCT CTATCCTGAT GGAACGAAAC 120
CTTCGAACTC CAAACCTGCT CGCATTCGTA AGGGCTGTCA AGCACTTAGA AAATGCATCA 180
GGTATTGAGC CAATTCTTTC TAATCTCCAA CCATGTGTTC CCTATTGGAC GGCCCCTCCC 240
TCTCGAACAC CAATCACGAT CAAGGCAGGT GACTGGCAAG AATTCCGGGA AAAACTGACG 300
TTCTATCTGG TTACCCTTGA GCAAGCGCAG GAACAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

398
ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA GCCACCGCTG 60
CCGCTGCTGG ACTTCAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGACAATAAC 120
CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA AGTCTCTGCA GAATGCATCA 180
GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC 240
ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG AATTCCGTCG TAAACTGACC 300
TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG 339

339 base pairs

nucleic acid

double

linear

DNA (genomic)

399
ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA GCCACCGCTG 60
CCGCTGCTGG ACTTCAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGAAAATAAC 120
CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA AGTCTCTGCA GAATGCATCA 180
GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCC 240
ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG AATTCCGTCG TAAACTGACC 300
TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG 339

29 base pairs

nucleic acid

double

linear

DNA (genomic)

400
AAGGAGATAT ATCCATGAAC TGCTCTAAC 29

5 amino acids

amino acid

linear

peptide

401
Met Asn Cys Ser Asn
1 5

120 amino acids

amino acid

linear

peptide

402
Met Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln
1 5 10 15
Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln
20 25 30
Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe
35 40 45
Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile
50 55 60
Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr
65 70 75 80
Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg
85 90 95
Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110
Thr Thr Leu Arg Leu Ala Ile Phe
115 120

14 amino acids

amino acid

linear

peptide

403
Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp Asp Lys
1 5 10

378 base pairs

nucleic acid

double

linear

DNA (genomic)

404
ATGGCATATC CAGAAACTGA TTACAAGGAC GACGATGACA AGGCTAACTG CTCTATAATG 60
ATCGATGAAA TTATACATCA CTTAAAGAGA CCACCTGCAC CTTTGCTGGA CCCGAACAAC 120
CTCAATGCTG AAGACGTCGA TATCCTGATG GAACGAAACC TTCGACTTCC AAACCTGGAG 180
AGCTTCGTAA GGGCTGTCAA GAACTTAGAA AATGCATCAG GTATTGAGGC AATTCTTCGT 240
AATCTCCAAC CATGTCTGCC CTCTGCCACG GCCGCACCCT CTCGACATCC AATCATCATC 300
AAGGCAGGTG ACTGGCAAGA ATTCCGGGAA AAACTGACGT TCTATCTGGT TACCCTTGAG 360
CAAGCGCAGG AACAACAG 378

378 base pairs

nucleic acid

double

linear

DNA (genomic)

405
ATGGCATATC CAGAAACTGA TTACAAGGAC GACGATGACA AGGCTAACTG CTCTATAATG 60
ATCGATGAAA TTATACATCA CTTAAAGAGA CCACCTAACC CTTTGCTGGA CCCGAACAAC 120
CTCAATTCCG AAGACATGGA TATCCTGATG GAACGAAACC TTCGAACTCC AAACCTGCTC 180
GCATTCGTAA GGGCTGTCAA GCACTTAGAA AATGCATCAG GTATTGAGGC AATTCTTCGT 240
AATCTCCAAC CATGTCTGCC CTCTGCCACG GCCGCACCCT CTCGACATCC AATCATCATC 300
AAGGCAGGTG ACTGGCAAGA ATTCCGGGAA AAACTGACGT TCTATCTGGT TACCCTTGAG 360
CAAGCGCAGG AACAACAG 378

113 amino acids

amino acid

linear

peptide

406
Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys
1 5 10 15
Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Asp Glu Asp
20 25 30
Met Ser Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala
35 40 45
Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser
50 55 60
Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro
65 70 75 80
Thr Arg His Pro Ile Ile Ile Arg Asp Gly Asp Trp Asn Glu Phe Arg
85 90 95
Arg Lys Leu Thr Phe Tyr Leu Trp Thr Leu Glu Asn Ala Gln Ala Gln
100 105 110
Gln

111 amino acids

amino acid

linear

peptide

407
Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro
1 5 10 15
Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp
20 25 30
Ile Leu Met Asp Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn
35 40 45
Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu
50 55 60
Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg
65 70 75 80
His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys
85 90 95
Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln
100 105 110

340 base pairs

nucleic acid

double

linear

DNA (genomic)

408
ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA GCCACCGCTG 60
CCGCTGCTGG ACTTCAACAA CCTCAATGGT GAAGACCAAG ATATCCTGAT GGAACAATAA 120
CCTTCGTCGT CCAAACCTCG AGGCATTCAA CCGTGCTGTC AACTCTCTGC AGAATGCATC 180
AGCAATTGAG AGCATTCTTA AAAATCTCCT GCCATGTCTG CCCCTGGCCA CGGCCGCACC 240
CACGCGACAT CCAATCCATA TCAAGGACGG TGACTGGAAT GAATTCCGTC GTAAACTGAC 300
CTTCTATCTG AAAACCTTGG AGAACGCGCA GGCTCAACAG 340

45 base pairs

nucleic acid

single

linear

DNA (synthetic)

409
CTTTAAGTGA TGTATAATTT CATCGATCAT TATAGAGCAG TTAGC 45

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

410
CACTTAAAGA GACCACCTGC ACCTTTGCTG GACCCG 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

411
GAGGTTGTTC GGGTCCAGCA AAGGTGCAGG TGGTCT 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

412
CACTTAAAGA GACCACCTAA CCCTTTGCTG GACCCG 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

413
GAGGTTGTTC GGGTCCAGCA AAGGGTTAGG TGGTCT 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

414
CACTTAAAGG TTCCACCTGC ACCTTTGCTG GACAGT 36

36 base pairs

nucleic acid

single

linear

DNA (synthetic)

415
GAGGTTGTTA CTGTCCAGCA AAGGTGCAGG TGGAAC 36

Number	Name	Date	Kind
4877729	Clark et al.	Oct 1989	A
4959455	Clark et al.	Sep 1990	A
5032395	Clark et al.	Jul 1991	A
5166322	Shaw et al.	Nov 1992	A
5591427	Vadas et al.	Jan 1997	A

Number	Date	Country
275598	Jul 1988	EP
413383	Feb 1991	EP
2210883	Jun 1989	GB
3236400	Oct 1991	JP
463595	Feb 1992	JP
8800598	Jan 1988	WO
8804691	Jun 1988	WO
8805469	Jul 1988	WO
8806161	Aug 1988	WO
9001039	Feb 1990	WO
9010705	Sep 1990	WO
9012874	Nov 1990	WO
9100350	Jan 1991	WO
9102754	Mar 1991	WO
9307171	Apr 1993	WO

	Number	Date	Country
Parent	07/981044	Nov 1992	US
Child	08/411795		US

Interleukin-3 (IL-3) multiple mutation polypeptides

Information

Patent Number

Date Filed

Date Issued

Inventors

Examiners

Agents

CPC

US Classifications

Field of Search

US

International Classifications

Disclaimer

Abstract

Description

Claims

Parent Case Info

US Referenced Citations (5)

Foreign Referenced Citations (15)

Non-Patent Literature Citations (14)

Continuation in Parts (1)