EUBACTERIUM RECTALE CAS12A MUTANTS

REFERENCE TO SEQUENCE LISTING

This application was filed with a Sequence Listing XML in ST.26 XML format accordance with 37 C.F.R. § 1.831 and PCT Rule 13ter. The Sequence Listing XML file submitted in the USPTO Patent Center, “013670-0028-WOO1_sequence_listing_xml_25 Apr. 2024.xml,” was created on Apr. 25, 2024, contains 1667 sequences, has a file size of 1.94 Mbytes (2,043,904 bytes), and is incorporated by reference in its entirety into the specification.

BACKGROUND

Eubacterium rectale Cas12a (ERCAS12A) is an engineered RNA-guided endonuclease from Eubacterium rectale (now called Agathobacter rectalis) Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) adaptive immune system. Zetsche et al., Cell 163: 759-771 (2015), which is incorporated by reference herein for such teachings. Cas12a is guided to a ˜21-24-nt DNA target sequence, or commonly referred as protospacer, by a target site-specific ˜21-24-nt complementary RNA. The Cas12a-gRNA ribonucleoprotein (RNP) complex mediates double-stranded DNA breaks (DSBs), which are then repaired by either the non-homologous end joining (NHEJ) which typically introduces mutations or indels at the cut site, or the homology directed repair (HDR) system for precise editing if a suitable template nucleic acid is present.

Critical to the recognition of correct DNA target for ERCAS12A variants include both crRNA and the canonical “TTTV” protospacer adjacent motif (PAM), which is a 4-bp sequence immediately upstream of the protospacer. Compared to the 2-bp NGG PAM of Cas9 from Streptococcus pyogenes, Cas12a expanded the targetable loci in genome editing, particularly over the T-rich sites that are inaccessible to the Cas9 system. Jinek et al., Science 337: 816-821 (2012), which is incorporated by reference herein for such teachings. Although the occurrence of targetable sites of Cas12a on the genome is less frequent than Cas9, ERCAS12A is an alternative enzyme for use in genome engineering applications. Improving the cleavage activity and associated genome editing efficiency of ERCAS12A would greatly facilitate the development of human cell therapies with enhanced properties.

Previous studies improved the utility of Cas12a from Lachnospiraceae bacterium ND2006 (LbCas12a) and Acidaminococcus sp. (AsCas12a). See, e.g., Gao et al., Nat. Biotechnol. 35(8): 789-792 (2017); Kleinstiver et al., Nat. Biotechnol. 37(3): 276-282 (2019); Zhang et al., Nat. Communications 12: 3908 (2021); U.S. Pat. App. Pub. Nos. US 2021/0348144 A1 and US 2023/0040148 A1, each of which are incorporated by reference herein for such teachings. However, transferring beneficial mutations from AsCas12a and LbCas12a to ERCAS12A was not simple or predictable. Despite significant structural similarity between these homologous proteins, most point mutations that improved the activity of AsCas12a and LbCas12a were detrimental to ERCAS12A.

What is needed are mutations that enhance the activity of ERCAS12A.

SUMMARY

One embodiment described herein is an isolated polypeptide comprising an Eubacterium rectale Cas12a (ERCAS12A) mutant having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant polypeptide provides an improvement in nuclease activity as compared to the wild-type ERCAS12A polypeptide. In one aspect, the ERCAS12A mutant has 98% to 99% identity to a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210). In another aspect, the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

Another embodiment described herein is an isolated polynucleotide sequence encoding an Eubacterium rectale Cas12a (ERCAS12A) mutant having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide. In one aspect, the encoded ERCAS12A mutant has 98% to 99% identity to a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the encoded ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant polynucleotide sequence has 95% to 99% identity to a nucleotide sequence selected from the odd numbered sequences of SEQ ID NO: 3-209. In another aspect, the ERCAS12A mutant polynucleotide has a nucleotide sequence selected from the odd numbered sequences of SEQ ID NO: 3-209. In another aspect, the encoded ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the encoded ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, encoded ERCAS12A mutant polynucleotide is selected from is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210). In another aspect, the ERCAS12A mutant poly nucleotide is selected from Q18V (SEQ ID NO: 3); 135L (SEQ ID NO: 5); G39Q (SEQ ID NO: 7); G39K (SEQ ID NO: 9); 140Q (SEQ ID NO: 11); D56R (SEQ ID NO: 13); D56P (SEQ ID NO: 15); D59R (SEQ ID NO: 17); D59A (SEQ ID NO: 19); S158G (SEQ ID NO: 21); K165Y (SEQ ID NO: 23); S181V (SEQ ID NO: 25); A290K (SEQ ID NO: 27); D291N (SEQ ID NO: 29); V296R (SEQ ID NO: 31); P297V (SEQ ID NO: 33); P297R (SEQ ID NO: 35); P297F (SEQ ID NO: 37); D303R (SEQ ID NO: 39); D303C (SEQ ID NO: 41); E305K (SEQ ID NO: 43); Y307K (SEQ ID NO: 45); Q353R (SEQ ID NO: 47); L444W (SEQ ID NO: 49); F465R (SEQ ID NO: 51); S469K (SEQ ID NO: 53); V470K (SEQ ID NO: 55); 1498K (SEQ ID NO: 57); Y507W (SEQ ID NO: 59); Y507S (SEQ ID NO: 61); Y507R (SEQ ID NO: 63); Q510Y (SEQ ID NO: 65); Q510V (SEQ ID NO: 67); Q51OK (SEQ ID NO: 69); P512V (SEQ ID NO: 71); P512G (SEQ ID NO: 73); Y513S (SEQ ID NO: 75); Y513R (SEQ ID NO: 77); Y513L (SEQ ID NO: 79); Y513G (SEQ ID NO: 81); S514R (SEQ ID NO: 83); S514P (SEQ ID NO: 85); T515P (SEQ ID NO: 87); K516R (SEQ ID NO: 89); K516P (SEQ ID NO: 91); K517P (SEQ ID NO: 93); 1518P (SEQ ID NO: 95); L520M (SEQ ID NO: 97); F522A (SEQ ID NO: 99); A541V (SEQ ID NO: 101); G553M (SEQ ID NO: 103); N556R (SEQ ID NO: 105); N556E (SEQ ID NO: 107); D562V (SEQ ID NO: 109); D562K (SEQ ID NO: 111); D5621 (SEQ ID NO: 113); N583V (SEQ ID NO: 115); L585K (SEQ ID NO: 117); F653R (SEQ ID NO: 119); K712R (SEQ ID NO: 121); N716R (SEQ ID NO: 123); E731P (SEQ ID NO: 125); N732T (SEQ ID NO: 127); A744T (SEQ ID NO: 129); A744P (SEQ ID NO: 131); A744N (SEQ ID NO: 132); F748A (SEQ ID NO: 135); S751A (SEQ ID NO: 137); K754G (SEQ ID NO: 139); 1757W (SEQ ID NO: 141); 1757K (SEQ ID NO: 143); E771R (SEQ ID NO: 145); N787A (SEQ ID NO: 147); A825R (SEQ ID NO: 149); T826K (SEQ ID NO: 151); T826F (SEQ ID NO: 153); Y832R (SEQ ID NO: 155); Y836Q (SEQ ID NO: 157); Y836K (SEQ ID NO: 159); Y839L (SEQ ID NO: 161); F840L (SEQ ID NO: 163); Y907V (SEQ ID NO: 165); Y9071 (SEQ ID NO: 167); Q916Y (SEQ ID NO: 169); E935R (SEQ ID NO: 171); 1936R (SEQ ID NO: 173); K974G (SEQ ID NO: 175); K1019V (SEQ ID NO: 177); 135L/S469K (SEQ ID NO: 179); 135L/T826F (SEQ ID NO: 181); 140Q/S469K (SEQ ID NO: 183); 140Q/T826F (SEQ ID NO: 185); S469K/K516P (SEQ ID NO: 187); S469K/T826F (SEQ ID NO: 189); Y513G/Y832R (SEQ ID NO: 191); Y513G/F840L (SEQ ID NO: 193); 135L/S469K/T826F (SEQ ID NO: 195); 140Q/S469K/T826F (SEQ ID NO: 197); S469K/K516P/T826F (SEQ ID NO: 199); S469K/T826F/F840L (SEQ ID NO: 201); D303R/S469K/T826F/F840L (SEQ ID NO: 203); F465R/S469K/T826F/F840L (SEQ ID NO: 205); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 207); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 209). In another aspect, the ERCAS12A mutant polynucleotide is selected from is selected from: D303R/S469K/T826F/F840L (SEQ ID NO: 203); F465R/S469K/T826F/F840L (SEQ ID NO: 205); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 207); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 209).

Another embodiment described herein is a vector or plasmid comprising the polynucleotide sequences described herein.

Another embodiment described herein is a cell comprising a nucleic acid comprising one or more of the polynucleotide sequences described herein.

Another embodiment described herein is an isolated CRISPR ribonucleoprotein complex comprising a guide RNA and a ERCAS12A mutant polypeptide having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide. In one aspect, the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210). In another aspect, the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

Another embodiment described herein is an isolated CRISPR/Cpf1 endonuclease system comprising a crRNA and a ERCAS12A mutant polypeptide having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2 where the ERCAS12A mutant polypeptide provides improved genome editing as compared to the wild-type ERCAS12A polypeptide. In one aspect, the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210). In another aspect, the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

Another embodiment described herein is a method for performing gene editing comprising: contacting a candidate editing target site locus with a CRISPR ribonucleoprotein complex comprising a crRNA and an ERCAS12A mutant polypeptide comprising having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides an improvement in gene editing as compared to the wild-type ERCAS12A polypeptide. In one aspect, the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant comprises ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210). In another aspect, the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

Another embodiment described herein is a kit comprising a CRISPR ribonucleoprotein complex comprising a guide RNA and an ERCAS12A mutant polypeptide comprising one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant polypeptide provides improved gene editing as compared to the wild-type ERCAS12A polypeptide. In one aspect, the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant comprises wherein the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q51OK, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2. In another aspect, the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210). In another aspect, the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

Another embodiment described herein is the use of a ERCAS12A mutant polypeptide for improving CRISPR/LbCas12a-associated nuclease activity at non-canonical TTTT PAM sites, wherein the ERCAS12A mutant one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide. In one aspect, the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210. In another aspect, the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q51OK (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

FIG. 1A-B show high-throughput characterization of ERCAS12A cleavage activity in E. coli. Three biological replicates of selection were performed and deep-sequenced to determine the enrichment of each ERCAS12A variant. FIG. 1A shows the phenotype scores of synonymous (n=834) and non-synonymous changes (n=22409) over the selection. Notably, most synonymous changes resulted in no apparent enrichment, serving as the baseline activity of wild-type ERCAS12A. FIG. 1B shows the enrichment scores for 23,243 ERCAS12A point mutations over two biological replicates. Results of the high-throughput screen are high reproducible (R²=0.66).

FIG. 2 shows DNA cleavage activity of ERCAS12A variants in E. coli. Each ERCAS12A mutant was cloned and evaluated in the context of E. coli. The survival rate of E. coli was determined by taking the ratio of colony counts with or without selection pressure. Most mutations resulted in higher survival rate than wild type (WT) ERCAS12A (˜10%). The raw data are shown in FIG. 3-5.

FIG. 3 shows examples of point mutations of ERCAS12A with enhanced DNA cleavage activity. Clearly, mutations increased the survival rate at TTTC and TTTT PAM sites, indicating an improved cleavage activity over the WT-ERCAS12A.

FIG. 4 shows examples of point mutations of ERCAS12A with enhanced DNA cleavage activity.

FIG. 5 shows examples of point mutations of ERCAS12A with enhanced DNA cleavage activity.

FIG. 6 shows cleavage activity of ERCAS12A variants in E. coli. Each ErCas12a mutant was cloned and evaluated in the context of E. coli. The survival rate of E. coli was determined by taking the ratio of colony counts with or without selection pressure. Most mutations resulted in higher survival rate than WT ERCAS12A (˜18%) FIG. 7A-D show the performance of ERCAS12A variants in human cells as RNP. ERCAS12A variant, and WT proteins were purified and assembled as RNPs using 6 different crRNAs. FIG. 7A-B show the editing efficiency of each variant measured by T7EI 48-hours post-delivery at 200 nM (FIG. 7A) or 50 nM RNP (FIG. 7B) in electroporation reaction. NC: negative control in the absence of ERCAS12A nuclease. FIG. 7C-D show normalized activity of each variant over WT ERCAS12A at 200 nM RNP (FIG. 7C) and 50 nM RNP (FIG. 7D) respectively. The dash line represents the baseline activity of WT protein.

FIG. 8A-C show the on-target editing efficiency of ERCAS12A point mutations in human cells. FIG. 8A shows the relative editing efficiency of ERCAS12A variants in human cells over two target sites. Plasmids encoding each variant were delivered by lipofection, and the editing efficiency was measured using a T7 endonuclease I assay (T7EI) 2 days post-delivery. FIG. 8B shows the editing efficiency of positive hits from lipofection experiment when delivered as RNP. FIG. 8C shows the normalized editing efficiency of ERCAS12A point mutations to WT.

FIG. 9A-B show the performance of stacked ERCAS12A variants with multiple point mutations in human cells as RNP. ERCAS12A variants, and WT proteins were purified and assembled as RNPs using 6 different crRNAs. FIG. 9A shows the editing efficiency of each variant measured by T7EI 48-hours post-delivery at 50 nM in electroporation reaction. NC: negative control in the absence of ERCAS12A nuclease. FIG. 9B shows the normalized activity of each variant over WT ERCAS12A at 50 nM RNP.

FIG. 10A-H shows the relative editing efficiency of ERCAS12A variants in human cells over 9 target sites. Plasmids encoding each variant were delivered by lipofection, and the editing efficiency was measured by NGS 3 days post-delivery. FIG. 10A shows AAVS1; FIG. 10B shows TGFBR2; FIG. 10C shows PD1; FIG. 10D shows TRAC1; FIG. 10D shows B2M-12; FIG. 10E shows CIITA; FIG. 10G shows CISH; and FIG. 10H shows TRAC2.

FIG. 11 shows the on-target editing efficiency of ERCAS12A Y513G in human cells. ERCAS12A Y513G, and WT proteins were purified and assembled as RNPs using 7 different crRNAs, and the editing efficiency was measured by NGS 2 days post-delivery.

FIG. 12A-D shows the on-target editing efficiency of ERCAS12A mutants in human cells. WT ERCAS12A, Y513G, S514P, Y513G/F840L and AsCas12a-Ultra (e.g., AsCas12a M537R/F870L) proteins were purified and assembled as RNPs using different crRNAs, and the editing efficiency was measured by NGS 2 days post-delivery. FIG. 12A shows AAVS1; FIG. 12B shows CISH; FIG. 12C showsTRAC1; FIG. 12D shows TRAC2.

FIG. 13 shows show the performance of stacked ERCAS12A variants with multiple point mutations in human cells as RNP. ERCAS12A variants, and WT proteins were purified and assembled as RNPs using different crRNAs. D303R/S469K/T826F/F840L; F465R/S469K/T826F/F840L; D303R/S469K/K516P/T826F/F840L) improved activity over ERCAS12A WT.

FIG. 14 shows a summary of the on-target editing efficiency of various ERCAS12A point mutations in human cells.

DETAILED DESCRIPTION

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For example, any nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are well known and commonly used in the art. In case of conflict, the present disclosure, including definitions, will control. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the embodiments and aspects described herein.

As used herein, the terms such as “include,” “including,” “contain,” “containing,” “having,” and the like mean “comprising.” The present disclosure also contemplates other embodiments “comprising,” “consisting of,” and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not.

As used herein, the term “a,” “an,” “the” and similar terms used in the context of the disclosure (especially in the context of the claims) are to be construed to cover both the singular and plural unless otherwise indicated herein or clearly contradicted by the context. In addition, “a,” “an,” or “the” means “one or more” unless otherwise specified.

As used herein, the term “or” can be conjunctive or disjunctive.

As used herein, the terms “amino acid,” “nucleotide,” “polynucleotide,” “vector,” “polypeptide,” and “protein” have their common meanings as would be understood by a biochemist of ordinary skill in the art. Standard single letter nucleotides (A, C, G, T, U) and standard single letter amino acids (A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y) are used herein.

As used herein, the terms “nucleic acid” and “polynucleotide” are interchangeable have the same meaning.

As used herein, the terms “amino acid sequence,” “polypeptide,” and “protein” are interchangeable have the same meaning.

As used herein, the term “substantially” means to a great or significant extent, but not completely.

As used herein, the term “about” or “approximately” as applied to one or more values of interest, refers to a value that is similar to a stated reference value, or within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, such as the limitations of the measurement system. In one aspect, the term “about” refers to any values, including both integers and fractional components that are within a variation of up to ±10% of the value modified by the term “about.” Alternatively, “about” can mean within 3 or more standard deviations, per the practice in the art. Alternatively, such as with respect to biological systems or processes, the term “about” can mean within an order of magnitude, in some embodiments within 5-fold, and in some embodiments within 2-fold, of a value. As used herein, the symbol “˜” means “about” or “approximately.”

All ranges disclosed herein include both end points as discrete values as well as all integers and fractions specified within the range. For example, a range of 0.1-2.0 includes 0.1, 0.2, 0.3, 0.4 . . . 2.0. If the end points are modified by the term “about,” the range specified is expanded by a variation of up to ±10% of any value within the range or within 3 or more standard deviations, including the end points.

As used herein, the terms “control,” or “reference” are used herein interchangeably. A “reference” or “control” level may be a predetermined value or range, which is employed as a baseline or benchmark against which to assess a measured result. “Control” also refers to control experiments or control cells.

As used herein, the phrase “an effective amount” of a compound described herein refers to an amount of the compound described herein that will elicit the biological response, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc.

As used herein, the terms “inhibit,” “inhibition,” or “inhibiting” refer to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

As used herein, the terms “Eubacterium rectale Cas12a” or “ERCAS12A” are used interchangeably and refer to the Eubacterium rectale (Er) Cas12a protein (previously named Cpf1), a class 2/type V CRISPR RNA-guided endonuclease.

The terms “ERCAS12A,” “wild-type ERCAS12A,” or “WT ERCAS12A” refer to a protein having the amino acid sequence of the naturally occurring Eubacterium rectale Cas12a (e.g., SEQ ID NO: 2, encoded by the nucleotide sequence of SEQ ID NO: 1) and that has biochemical activity when combined with a suitable crRNA to form an active CRISPR/Cas12a endonuclease system.

The term “mutant ERCAS12A protein” or “variant ERCAS12A protein” are used interchangeably and refer to Eubacterium rectale Cas12a protein forms having a different amino acid sequence form the wild-type Eubacterium rectale Cas12a proteins and that have biochemical activity when combined with a suitable crRNA to form an active CRISPR-Cas12a endonuclease system. This includes orthologs and Cas12a variants having different amino acid sequences from the wild-type Eubacterium rectale Cas12a. The mutant ERCAS12A proteins described herein have at least one amino acid substitution to the wild-type ERCAS12A polypeptide sequence. In one aspect, the mutant ERCAS12A polypeptides have at least one amino acid substitution to the wild-type ERCAS12A polypeptide sequence and have an improvement in CRISPR/ERCAS12A-associated nuclease activity at non-canonical TTTT PAM sites as compared to the wild-type ERCAS12A enzyme.

As used herein, the term “substantially purified,” as applied to a composition, refers to a composition having at least 90% purity or greater, including 90% purity, 95% purity, 99% purity and greater than 99% purity.

As used herein, the term “isolated,” when used to describe a composition, such as a polynucleotide, a polypeptide or a ribonucleoprotein complex refers to a substantially purified composition, or in the case of a ribonucleoprotein complex, at least one component being a substantially purified component. In further respect to an isolated ribonucleoprotein complex, preferably all components are substantially purified.

As used herein, the term “affinity tag” refers to a ligand that permits detection and/or selection of an oligonucleotide sequence to which the ligand is attached. For the purposes of this disclosure, a bait may include an affinity tag. In particular, the affinity tag is positioned typically at either or both the N-terminus and/or C-terminus of a polypeptide through conventional chemical coupling technology or recombinant DNA technology. Exemplary affinity tags include biotin, digoxigenin, streptavidin, polyhistine, e.g., (His₆), glutathione-S-transferase (GST), HaloTag®, AviTag, Calmodulin-tag, polyglutamate tag, FLAG-tag, HA-tag, Myc-tag, S-tag, SBP-tag, Softag 3, V5 tag, Xpress tag, a hapten, among others.

As used herein, the term “eukaryotic cell,” includes those cells of or derived from a particular organism, such as a plant or a mammal, including but not limited to human, or non-human eukaryote or animal or mammal as herein discussed, e.g., mouse, rat, rabbit, dog, livestock, or non-human mammal or primate. In some embodiments, processes for modifying the germ line genetic identity of human beings and/or processes for modifying the genetic identity of animals which are likely to cause them suffering without any substantial medical benefit to man or animal, and also animals resulting from such processes, may be excluded. Preferred human cells include cells derived from somatic cells and germ line cells. Exemplary somatic cells include cells from every major organ and tissue system, including the immune system and hematopoietic system.

As used herein, the phrase “the odd numbered sequences of SEQ ID NO: 3-209” refers to nucleotide sequences of SEQ ID NO: 3, 5, 7 . . . 207, or 209. See Table 14, where all odd numbered sequences are nucleotide sequences.

As used herein, the phrase “the even numbered sequences of SEQ ID NO: 4-210” refers to polypeptide sequences of SEQ ID NO: 4, 6, 8 . . . 206, or 210. See Table 14, where all even numbered sequences are polypeptide sequences.

Described herein are Eubacterium rectale Cas12a (ERCAS12A) variants and methods to enhance the utility of ERCAS12A and variants thereof.

A bacterial-based directed evolution of ERCAS12A was used to identify mutations with enhanced cleavage activity. First a deep-scanning mutagenesis library containing all possible point mutations on the amino acid level over the entire coding sequence of ERCAS12A was created. Most clones contained only one mutation. This type of library allowed us to directly evaluate the phenotype of each point mutation, by measuring their relative survival rates over the wild-type (WT) protein in the bacterial screen. Briefly, the screening strain harboring the toxin plasmid was transformed with ERCAS12A library and crRNA targeting the HPRT38346 site on the toxin plasmid. After recovery and IPTG induction, cells were plated on LB-Chloramphenicol media with arabinose and incubated at 37° C. overnight. ERCAS12A expression plasmids carried by the survived E. coli cells were extracted and purified. Both input and selected plasmid libraries were PCR amplified, randomly fragmented by Nextera library prep kit, and sequenced on Illumina NextSeq with ˜40 million reads per library. The frequencies of mutations at each position of ERCAS12A in both libraries were measured and normalized to the total coverage of each codon. The relative survival rate of each point mutation was calculated as the ratio of normalized frequency between selected and input library. Since the degree of cell survival under the arabinose selection is indicative of the cleavage activity of ERCAS12A variant at HPRT38346 protospacer, any variants that enriched during the selection over WT would be those with enhanced activity at TTTT PAM.

Large collection of putative mutations with enhanced activity in the context of E. coli is described. The phenotype of 22,049 point mutations of ERCAS12A in the bacterial screening that measuring the cleavage activity at non-canonical TTTT PAM. Two biological replicates were performed with phenotype measurements which enabled the ability to isolate a large collection of ERCAS12A variants with enhanced cleavage activity.

The genome editing efficiency of ERCAS12A variants with single or multiple point mutations in human cells were evaluated. ERCAS12A variants were column purified, and delivered to human HEK293 cells as ribonucleoprotein (RNP) to edit 6 genomic targets with canonical TTTV PAM. Editing efficiency of each mutant was quantified by T7EI assay and compared to WT ERCAS12A. This enabled rigorous verification of the activity of putative candidates in the context of human cells without protein overexpression. The editing efficiency was measured by T7 endonuclease I assay (T7E1) 72 hours post-delivery.

The genome editing efficiency of 10 ERCAS12A variants with single or multiple point mutations in human cells. ERCAS12A variants were column purified, and delivered to human HEK293 cells as ribonucleoprotein (RNP) to edit 6 genomic targets with canonical TTTV PAM. Editing efficiency of each mutant was quantified by T7EI assay and compared to WT ERCAS12A. This enabled rigorous verification of the activity of putative candidates in the context of human cells without protein overexpression. The editing efficiency was measured by T7 endonuclease I assay (T7EI) 72-hours post-delivery.

One embodiment described herein is a CRISPR-associated protein comprising a polypeptide encoding a variant of ERCAS12A. The isolated polypeptide comprising a ERCAS12A variant is selected from a polypeptide having one or more amino acid substitutions presented in Table 2 relative to wild type ERCAS12A (SEQ ID NO: 2).

Another embodiment described herein is an ErCas12a variant selected from a polypeptide a polypeptide having one or more amino acid substitutions presented in Table 2 relative to wild type ERCAS12A (SEQ ID NO: 2), wherein the selected ERCAS12A variant displays improved genome editing relative to wild type ERCAS12A (SEQ ID NO: 2), wherein the selected ERCAS12A variant displays improved genome editing relative to wild type ERCAS12A (SEQ ID NO: 2).

Another embodiment described herein is CRISPR ribonucleoprotein complex is provided. The CRISPR ribonucleoprotein including a polypeptide encoding a variant of ERCAS12A. The variant of ERCAS12A is selected from the ERCAS12A variant is selected from a polypeptide having one or more amino acid substitutions presented in Table 2 relative to wild type ERCAS12A (SEQ ID NO: 2).

Another embodiment described herein is method of increasing efficiency of gene editing at TTTN PAM sites in a cell with a CRISPR ribonucleoprotein complex is provided. The method includes the step of contacting a cell with the CRISPR ribonucleoprotein complex. The CRISPR ribonucleoprotein complex includes a guide RNA and a CRISPR-associated protein. The CRISPR-associated protein including a polypeptide encoding a variant of ERCAS12A. The variant of ERCAS12A is selected from the ERCAS12A variant is selected from a polypeptide having one or more amino acid substitutions presented in Table 2 relative to wild type ERCAS12A (SEQ ID NO: 2).

Another embodiment described herein is a kit comprising a guide RNA and a CRISPR-associated protein is provided. The CRISPR-associated protein includes a polypeptide encoding a variant of ERCAS12A.

Another embodiment described herein is a CRISPR-associated protein comprising a polypeptide encoding a variant of ERCAS12A, wherein comprises at least one variant amino acid selected from amino acid positions M1G, N2C, N31, T5D, T5F, T5V, N6P, N7A, N7D, N7E, N71, N7Q, N7Y, F8H, F8K, Q9D, N10A, N10E, F11E, F11Q, 112L, 112Q, 114E, 114H, 114L, 114N, 114V, S15K, S15M, S15T, S16H, S16M, S16N, Q181, Q18M, Q18N, Q18R, Q18T, Q18V, Q18W, A24E, 126P, P27C, P27F, P27G, P27H, P27K, P27L, P27M, P27R, P27Y, T28G, T30D, T30F, T30H, T30Q, T30V, T30W, T31A, T31C, T31D, T31G, T31H, T31L, T31M, T31N, T31P, T31S, Q32A, Q32C, Q32F, Q32G, Q321, Q32K, Q32L, Q32M, Q32N, Q32P, Q32S, Q32T, Q32V, Q32W, Q32Y, Q33F, Q331, Q33P, Q33V, Q33W, Q33Y, F34A, F34L, F34P, F34R, F34W, 135A, 135C, 135E, 135F, 135G, 135K, 135L, 135M, 135N, 135P, 135Q, 135S, 135T, 135V, 135W, 135Y, V36K, V36M, V36P, K37P, N38A, N38D, N38E, N38G, N38H, N381, N38L, N38M, N38P, N38Q, N38R, N38S, N38T, N38V, G39A, G39C, G39D, G39E, G39F, G39H, G391, G39K, G39L, G39M, G39N, G39P, G39Q, G39R, G39S, G39T, G39V, G39W, G39Y, 140C, 140D, 140E, 140K, 140L, 140N, 140P, 140Q, 140R, 140T, 140V, 140W, 140Y, E43C, E431, E43L, E43Q, E43S, E43T, E43V, E45F, E45R, L46C, L46D, L46E, L46G, L46H, L46K, L46N, L46S, L46T, E49C, E49F, E49H, E491, E49K, E49L, E49M, E49N, E49P, E49R, E49S, E49T, E49Y, N50Q, Q52C, Q52V, L54G, D56A, D56K, D56P, D56R, 157A, 157C, 157E, 157G, 157L, 157P, 157Q, 157T, 157V, D59A, D59G, D591, D59L, D59M, D59Q, D59R, D59S, D59T, D60A, D60F, D60G, D60K, D60L, D60M, D60N, D60P, D60R, D60T, D60V, D60Y, Y62A, Y62F, Y62N, Y62Q, Y62V, R63M, R63V, F65E, F65K, F65R, S67A, S67F, S67Q, S67V, S67W, S67Y, E68A, E68R, E68V, T69D, T69F, T69H, T69L, T69M, T69W, S71Q, 173E, D75R, 176F, 176G, 176Q, D77P, T79E, T79R, S80N, S80Q, S80V, L81E, L81H, L81W, L81Y, F82Q, K84F, M85A, M85E, M85F, M85G, M85N, M85Q, M85W, M85Y, E86N, 187A, 187G, 187H, 187W, Q88E, Q88N, Q88S, L89C, L89D, L89G, L89Q, L89S, K90P, N91V, G92D, G92F, D93L, D93M, D93N, D93P, D93R, D93T, N94P, K95E, K95F, K951, K95L, K95P, K95S, K95T, K95W, D96C, T97W, L98A, L98C, L98D, L98G, L98H, L98N, L98P, L98Q, L98S, L98T, 199W, 199Y, E101P, Q1021, Q102V, T103Y, E104P, Y105A, Y105G, Y105P, Y105R, Y105T, R1061, R106T, R106V, A108R, K111A, K111C, K111F, K111G, K111S, K111T, K111V, K111W, K112C, K112D, K112E, K112H, K112L, K112P, K112R, K112T, F113A, A114E, A114S, N115M, N115W, R118A, R118D, R118N, R118V, R118Y, F119D, F119H, K120A, K120C, K120P, K120T, K120V, N121F, M122W, F123E, S124N, L127Q, 1128Q, S129D, S129F, S129G, S1291, D1301, L132A, L132G, L132T, F135A, F135D, F135H, F135T, V136Y, 1137Q, H138A, H138M, H138S, H138V, N139A, N139C, N139F, N139G, N1391, N139L, N139P, N139Q, N139R, N140C, N140G, N140P, N140S, N140T, N141F, N141P, Y142E, Y142G, Y142K, Y142P, Y142R, Y142V, S143E, S143G, S1431, S143R, S143V, A144C, A144D, A1441, A144K, A144N, A144P, A144R, A144T, E1461, E146M, E146R, E146S, E146T, K147A, K147F, K147G, K147L, K147Q, E148C, E148P, E149L, E149R, K150A, K150C, K150G, K1501, K150L, K150V, K150W, K150Y, T151C, T151E, T151Q, T151V, V153K, 1154A, 1154E, 1154H, 1154P, 1154Q, L156C, S158D, S158G, S158K, S158N, F160H, S1631, F164Q, K165Y, Y1671, Y167L, Y167M, K169C, K169F, A172Y, C1741, C174T, C174V, A177M, A177W, D178R, D179P, D179W, S181H, S181N, S181V, S184C, S1841, S184L, S184M, S184V, 1188A, 1188C, 1188W, N190Q, N190R, N192C, N192G, A193F, A193H, A193L, A193N, A193Q, 1195W, F196T, F197E, F197H, F197K, F197Q, F197R, S198D, S198E, S198K, S198N, A200H, A2001, V202C, V202H, R205C, R205M, R205T, 1206D, 1206W, 1206Y, V207W, S209K, L210N, D214T, D214W, 1215A, 1215W, N216F, K217A, K217D, K217V, S219D, S219F, S219P, S219W, G220Y, D221C, D2211, D221L, D221N, D221P, D221R, D221S, M222C, M222P, K223C, K223F, K223G, K223P, D224H, S225A, S225W, S225Y, L226D, L226Y, E228F, E228P, E228T, E228W, M229S, S230E, S230G, L231D, L231K, L231R, E233S, 1234G, 1234W, Y235A, Y235L, Y235N, Y235Q, Y235T, Y235W, S236G, S236K, S236N, S236V, S236W, E238F, E238S, K239F, K239G, K239T, K239V, K239W, G241F, G241H, G241M, G241P, G241Q, G241W, F243H, F243R, 1244F, 1244G, 1244Q, 1244W, 1244Y, T245E, T245K, T245L, T245Q, T245V, T245Y, E247C, E2471, E247M, E247T, G248K, G248M, S250N, N253A, N253G, N253Q, N253W, D2541, D254M, D254Q, D254R, D254S, D254W, C256E, C256H, C256M, C256N, C256Q, K258L, V259N, V259Q, V259Y, S261F, S261H, F262G, F262K, F262P, F262R, M263C, M263E, M263S, L265K, L265W, Y266E, Y266P, Y266R, C267E, C267K, C267M, C267P, C267T, Q268C, Q268G, Q268V, K269Y, N270E, N270G, K271C, K271H, K2711, K271W, E272G, E272K, E272P, E272S, E272Y, N2731, N273R, N273S, N275G, L276K, L276N, Y277L, Y277M, Y277P, K278A, K278F, K278L, K278P, K278S, K278W, Q280D, Q280M, K281D, K281F, K281G, L282K, L282S, H283M, Q285M, Q285T, 1286C, C288G, C288K, 1289W, A2901, A290K, A290N, A290R, D291F, D291L, D291M, D291N, D291P, D291T, D291W, T292H, S293H, S293K, S293W, Y294L, Y294M, Y294V, E295C, E295M, E295R, E295T, V296A, V296D, V296G, V296H, V296K, V296M, V296N, V296P, V296Q, V296R, V296T, V296W, V296Y, P297C, P297F, P297H, P2971, P297K, P297M, P297N, P297R, P297V, P297W, P297Y, K299A, K299E, K299G, K299P, K299V, K299W, F300D, F300E, F300K, F300M, F300Q, E301A, E301C, E301F, E301G, E301K, E301M, E301P, E301Q, E301R, E301T, E301W, S302A, S302C, S302D, S302E, S302G, S302H, S3021, S302M, S302N, S302V, S302W, D303C, D303F, D303G, D303H, D3031, D303K, D303M, D303N, D303P, D303Q, D303R, D303S, D303V, D303W, D303Y, E304T, E305A, E305C, E305F, E305G, E305H, E3051, E305K, E305L, E305M, E305N, E305P, E305R, E305S, E305V, E305W, E305Y, V306C, V306F, V306W, V306Y, Y307A, Y307D, Y307E, Y307G, Y3071, Y307K, Y307L, Y307N, Y307P, Y307Q, Y307R, Y307S, Y307T, Y307V, Y307W, Q3081, Q308W, S309K, S309L, S309M, S309V, V310Y, N311F, N311L, G312A, G312E, G3121, G312K, G312Q, G312S, G312T, F313H, F313M, D3151, D315M, 1317W, S318D, S318K, S3191, S319K, S319N, K320F, H321F, H321V, 1322A, E324C, E324M, E324T, E324W, E324Y, R325K, R327K, R327M, R327V, D331C, D331M, Y333M, N334W, G335Q, Y336H, Y336M, Y336V, Y336W, N337G, D339M, D339Q, D339S, K340A, K340E, K340H, Y342M, Y342R, 1343V, V344E, K346C, F347G, F347R, Y348M, Y348V, E349K, E349N, E349Q, E349R, E349T, S350H, S350R, V351D, V351F, V351K, V351Y, S352M, Q353K, Q353M, Q353N, Q353R, T355M, Y356A, Y356C, Y356E, Y356G, Y3561, Y356M, Y356V, R357K, R357M, D358R, D358W, T361K, T364W, A365N, A365Y, E367M, E367P, 1368P, 1368Y, H369A, H369E, H369G, H3691, H369V, Y370A, Y370G, Y370N, Y370P, Y370R, N371C, N371G, N372A, N372F, N372W, 1373D, L374E, L374H, L374N, L374Y, P3751, P375Y, G376Q, N377L, N377P, N377V, G378N, G378W, K379A, K379F, K379G, K379R, K381L, K381P, K381W, D383M, D383P, K384D, K384G, K384P, V385C, V385Y, K386F, A388H, A3881, A388N, A388Y, V389E, K390C, N391W, N391Y, L393D, L393Y, Q394A, Q394C, Q394F, Q394G, Q3941, Q394M, Q394S, Q394T, Q394V, Q394W, K395F, K3951, K395V, 1397C, 1397Q, T398D, T398F, E399M, 1400E, E402H, S405K, S405M, S405R, N406A, N406V, Y4071, Y407P, Y407R, K408C, K408D, K4081, C410F, S411N, S411V, D413R, D413T, D413W, N414A, N414D, N414H, N414L, N414S, 1415C, 1415E, K416P, K416V, K416W, A417C, A417M, E418W, T419G, T419H, T419Y, Y420K, Y420Q, Y420R, Y420V, 1421 F, H422A, H422C, H422F, H422M, E423A, E423H, E423R, E423S, E423T, E423Y, S425E, S425K, H426G, H4261, 1427H, L428T, L428Y, N429A, N429P, F431A, F431R, E432C, E432L, E432R, E432T, Q434F, Q4341, Q434L, Q434P, Q434S, Q434T, Q434W, E435A, E435L, E435Y, L436A, L436D, L436E, L436G, L436K, L436P, L436R, L436S, L436T, L436W, K437C, K437H, K437P, K437W, Y438A, Y438E, Y438G, Y4381, Y438K, Y438L, Y438P, Y438R, Y438W, N439A, N439G, N439H, N439P, N439S, P440A, P440D, P440E, P440G, P440K, P440L, P440M, P440N, P440Q, P440R, P440S, P440T, E441F, E441G, E441H, E441K, E441N, E441R, 1442A, 1442G, 1442P, 1442Y, H443A, H443E, H443F, H443W, L444A, L444C, L444D, L444E, L444G, L444H, L4441, L444K, L444N, L444T, L444W, L444Y, V445C, V445E, V445K, V445P, V445Q, V445R, V445W, E446P, E446T, E446W, S447D, S447F, S4471, S447L, S447P, S447Q, S447W, S447Y, E448A, E448H, E4481, E448W, E448Y, L449D, K450C, K450G, K450S, K450V, A451C, A4511, A451M, S452G, S452P, S452V, E453F, E453W, L454R, L454S, L454Y, N456M, N456P, N456Q, N456R, V457E, V457T, V457Y, V460E, V460H, V4601, V460K, V460N, V460P, 1461C, 1461Y, N463A, F465A, F465E, F465G, F465K, F465N, F465Q, F465R, F465S, F465T, F465V, H466C, H466K, S469K, S469L, S469M, S469P, S469R, S469V, V470K, V470N, V470Y, M472G, T473F, T473G, T473R, E474P, L476G, L476H, L476K, L476N, L476R, L476T, V477D, V477E, V477M, V477Q, V477T, V477W, K479A, K479D, K479H, D480P, D480Q, N481A, N481D, N482A, N482D, N482H, N482Q, N482S, F483A, F483W, Y484G, Y484R, Y484V, A485H, A485K, A485L, A485Q, A485R, A485S, E486A, E486F, E486G, E486L, E486R, E486S, E486Y, E488Q, E488T, E489C, E489H, E489M, E489S, E489T, E489W, 1490A, 1490E, Y491E, Y491Q, D4921, D492K, D492L, E493C, E493R, Y495V, P496C, P496E, P4961, P496K, P496L, P496S, P496V, P496W, V497W, 1498G, 1498K, 1498P, 1498R, S499C, S499G, S499M, L500S, L503D, L503E, L503G, L503H, L5031, L503K, L503N, L503P, L503R, L503W, V504A, V504C, V504G, V504T, Y507A, Y507C, Y507E, Y507G, Y5071, Y507K, Y507L, Y507Q, Y507R, Y507S, Y507T, Y507V, Y507W, T509L, T509M, T509V, Q51OA, Q510C, Q510D, Q51OE, Q51OF, Q510G, Q510H, Q5101, Q510K, Q510M, Q510N, Q510P, Q510R, Q510S, Q510T, Q510V, Q51OW, Q510Y, K511F, K511G, K511S, K511V, K511Y, P512C, P512D, P512E, P512F, P512G, P512H, P5121, P512N, P512R, P512S, P512T, P512V, P512W, P512Y, Y513A, Y513C, Y513D, Y513E, Y513G, Y513H, Y5131, Y513K, Y513L, Y513M, Y513N, Y513P, Y513Q, Y513R, Y513S, Y513T, Y513V, S514A, S514D, S514F, S514G, S514H, S514K, S514L, S514N, S514P, S514R, S514W, T515A, T515C, T515G, T515L, T515M, T515P, T515S, T515Y, K516A, K516P, K516R, K516S, K516T, K517A, K517D, K517G, K517L, K517P, 1518E, 1518F, 1518L, 1518P, 1518Q, 1518S, 1518T, L520A, L520G, L520H, L520M, L520N, L520Q, L520S, N521E, N521F, N521L, N521P, N521R, F522A, F522C, F522G, F522M, G523A, G523Q, A5281, Y537F, S538C, S538N, A5411, A541L, A541M, A541N, A541Q, A541T, A541V, 1543A, 1543C, 1543F, 1543G, 1543H, 1543Q, 1543S, 1543T, 1543W, L544W, L544Y, M545E, M545Q, R546F, L549D, L552E, L552K, L552Q, G553A, G5531, G553L, G553M, G553S, G553T, G553V, 1554C, F555M, N556A, N556C, N556E, N556F, N556K, N556M, N556Q, N556R, N556V, N556W, A557K, A557S, K558E, K558F, K558H, K558M, K558S, K558T, N559A, N559E, N559G, D562C, D562F, D5621, D562K, D562L, D562M, D562Q, D562T, D562V, D562W, K564A, K564D, K564G, 1565F, 1565R, 1566A, 1566W, E567L, E567Q, E567W, N569P, N569W, T570D, T570F, S571M, S571Q, S571W, S571Y, E572M, K574F, K5741, K574V, G575D, G575W, Y577M, Y577P, K578A, K578C, K578F, M580Q, 1581H, N583A, N583G, N5831, N583S, N583T, N583V, L585K, P586F, G587C, G587H, G602A, E6041, E604T, T605C, T605G, K607C, S609C, S609E, S609K, A610H, A6101, A610N, A610Y, L613G, E6141, K617C, K617Q, K617V, Q618D, N619C, N619R, K620F, S624W, S625N, K6261, K626V, K626Y, H634K, L636H, D638C, D6381, D638S, Y639G, Y639H, Y639M, Y639N, Y639T, N642A, N642M, C643F, C643T, 1644A, 1644Y, A645M, A645N, A645Q, P648C, P6481, P648M, F653R, G654P, F655H, D656G, D656K, S658D, D659K, S661G, S661M, S661Q, E664F, E664R, E664S, S667R, G668M, G668Q, F669P, F669R, L675C, Q676S, Q676T, Q676V, Q676W, Y678F, Y678H, 1680A, 1680F, 1680H, 1680N, 1680Q, 1680S, 1680V, D6811, D681K, D681M, D681Q, D681R, D681T, D681W, W682A, W682D, W682E, W682G, W6821, W682K, W682M, W682N, W682Q, W682T, W682V, T683F, T683Y, 1685Y, S686R, S686T, E687N, K688D, K688V, D691R, L692M, L693A, L693D, L693E, L693G, L693M, L693Q, L693T, L693W, Q694C, Q694G, Q6941, Q694K, Q694R, Q694S, Q694W, Q694Y, E695S, G6971, G697N, G697P, Q698A, Q698F, L699C, L699F, L699Y, Y700A, Y700G, Y7001, Y700T, Y700V, L701P, L701W, F702A, F702C, F702G, F702H, F702L, F702M, F702N, F702Q, F702S, F702T, F702V, Q703C, K711L, K711P, K712A, K712C, K712F, K712G, K712H, K712Q, K712R, K712S, K712W, K712Y, S713R, T714N, T714W, T714Y, N716A, N716C, N7161, N716K, N716M, N716Q, N716R, N716T, N716V, D717A, D717P, L719K, K725Y, L727H, L7271, L727T, F728T, S729H, S729K, E730G, E7301, E730L, E730M, E730V, E731A, E731L, E731P, N732D, N732T, L7331, L733K, D735K, D735R, V737Q, L738E, N741C, N741G, E743N, A744C, A744N, A744P, A744S, A744T, E745P, F748A, F748C, F748D, F748E, F748G, F748H, F7481, F748M, F748N, F748Q, F748R, F748S, F748T, F748V, F748W, K750A, K750E, K750F, K750G, K750H, K750L, K750N, K750P, K750S, K750T, K750W, K750Y, S751A, S751C, S751D, S751E, S751F, S751G, S751H, S751L, S751M, S751N, S751P, S751Q, S751R, S751T, S751V, S752C, S752F, S752G, S752H, S7521, S752L, S752M, S752Q, S752V, S752W, S752Y, 1753E, K754A, K754C, K754D, K754E, K754F, K754G, K754H, K7541, K754L, K754N, K754P, K754Q, K754S, K754T, K754V, K754W, K754Y, P756H, P7561, P756K, P756L, P756M, P756N, P756S, P756T, 1757A, 1757C, 1757F, 1757G, 1757K, 1757L, 1757M, 1757P, 1757Q, 1757R, 1757S, 1757W, 1757Y, 1758A, 1758C, 1758E, 1758G, 1758K, 1758Q, 1758S, 1758T, 1758V, G762K, 1764Q, V766Y, N767R, R768K, T769L, E771A, E771C, E771F, E771H, E7711, E771K, E771M, E771Q, E771R, E771S, E771T, E771V, E771W, E771Y, A772W, E7731, E773P, E774H, E7741, E774K, E774L, E774M, E774Q, E774R, E774S, E774T, E774V, E774W, K775C, Q777R, Q777V, F778G, F778K, F778Q, F778R, 1783Q, V784F, V784H, V7841, V784L, V784R, V784W, V784Y, N787A, N787C, N787F, N787G, N787H, N787M, N787P, N787Q, N787R, N787S, N787W, 1788F, 1788Y, P789C, E790A, E790C, E790F, E790G, E790H, E7901, E790K, E790M, E790N, E790P, E790Q, E790R, E790S, E790T, E790V, E790W, E790Y, N791E, N791F, N791G, Y793Q, Y793R, Y793V, Y793W, Q794L, Q794P, Y797W, K798L, F800W, N801A, N801S, K803A, S804F, S8041, K806C, S809G, A813H, E823A, E823P, E823S, A825H, A825K, A825R, T826C, T826F, T826H, T8261, T826K, T826L, T826M, T826N, T826Q, T826R, T826W, T826Y, N827D, N827E, V829T, D831L, D831M, D831N, D831Q, D831R, Y832A, Y832E, Y832G, Y832K, Y832M, Y832N, Y832Q, Y832R, Y836A, Y836C, Y836D, Y836E, Y836G, Y836H, Y8361, Y836K, Y836L, Y836M, Y836N, Y836Q, Y836R, Y836S, Y836T, Y836V, D837P, Y839A, Y839C, Y839H, Y8391, Y839K, Y839L, Y839M, Y839N, Y839Q, Y839R, Y839S, Y839T, Y839V, L841C, L841G, L8411, L841V, L841Y, M843F, M843Q, P844C, 1845Q, T846G, T846L, T846Q, T846W, T846Y, N848D, F849R, A851N, A851T, 1857A, 1857N, D859P, R860E, R860K, 1861A, L862D, L862H, L862Y, A866D, A866Y, K869C, H872M, H872S, 1876L, V886A, V886G, V886S, T8911, C892D, C892H, C8921, C892T, G893K, N894V, E897Y, Q898G, F901G, F901L, F901M, F901Q, 1903V, Y9071, Y907M, Y907T, Y907V, Y909F, Y909W, Q91OA, Q91ON, Q910S, Q910Y, 1911A, 1911D, 1911E, 1911G, 1911H, 1911Q, 1911W, 1911Y, K912C, K912H, K912Y, K914A, K914C, K914F, K914M, K914Q, K914S, K914T, Q916A, Q916D, Q916L, Q916M, Q916N, Q916V, Q916W, Q916Y, G918D, G918E, G918F, G918H, G9181, G918K, G918L, G918M, G918Q, G918T, G918Y, A919E, A919F, A9191, A919L, A919M, A919Y, Q921A, Q921T, 1922A, 1922C, 1922H, 1922R, 1922W, 1922Y, K925C, E926N, E926S, E926T, K928W, E929C, E929G, E929N, E929P, E929S, G931H, G931Q, K934S, E935A, E935C, E935F, E935G, E935H, E935K, E935L, E935M, E935N, E935Q, E935R, E935S, E935T, E935W, E935Y, 1936A, 1936C, 1936F, 1936G, 1936K, 1936L, 1936Q, 1936R, 1936S, 1936T, 1936Y, L941C, L941N, S942A, L9431, V944C, 1945M, H946W, E947F, E947M, E947P, 1948L, S9491, S949K, S949M, S949Q, S949T, S949V, M951G, V952E, V952W, 1953A, 1953P, K954A, K954P, Y955C, Y955W, N956C, N956F, N956W, N956Y, A957F, A957P, A957T, 1958V, 1959F, D963C, F968W, K970C, K974A, K974C, K974E, K974G, K974M, K974P, K974Q, K974S, K974T, K974W, K974Y, V9751, R977H, K982S, K982Y, F983A, M986G, 1988E, 1988K, L991C, L991W, N992A, N992L, L994A, V995P, F996T, F996V, D998T, D998W, 1999H, 1999P, 11001W, T1002C, T1002L, T1002S, T1002V, E1003M, E1003S, N1004A, N1004G, N1004P, N1004Q, N1004R, L1007A, L1007C, L1007V, L1008F, Y10111, Y1011L, Y1011V, T1014C, Y1015A, Y1015E, Y1015G, Y1015M, Y1015Q, Y1015T, 11016A, 11016E, 11016N, 11016P, P1017D, P1017E, P1017G, P1017N, P1017Q, P1017W, K1019A, K1019C, K1019G, K1019S, K1019V, K1021A, K1021F, K1021V, K1021W, V1023C, V1023N, G1024Y, Q1026M, Q1026N, C1027N, C1029N, Y1032K, Y1032Q, Y1032R, Y1032W, A1036F, A1036H, K1040A, K10401, K1040T, 11050D, 11050H, 11050W, K1052C, K1052G, K10521, K1052W, F1053Y, K1054F, K10541, K1054L, K1054M, K1054V, K1054W, D1055C, D1055F, D10551, D1055L, D1055M, D1055P, D1055Q, D1055T, D1055V, L1056C, L1056E, L10561, T1057Y, V1058H, V1058W, A1060N, A1060Q, K1061C, K1061D, K1061E, K1061F, K10611, K1061L, K1061Q, K1061R, K1061S, K1061T, K1061W, R1062A, R1062D, R1062H, R1062M, R1062V, R1062Y, E1063F, E1063G, E1063W, 11065W, F1068N, D1069M, S1070E, R1072D, R1072E, D1074R, D1074S, S1075V, E1076T, K1077R, C1081A, C10811, T1083E, T1083K, T1083M, T1083Y, N1087A, N1087W, N1088A, N1088P, 11090G, T1091C, T1091W, Q1092A, Q1092C, Q1092F, Q10921, Q1092Y, N1093C, T10941, T1094P, T1094Y, V1095A, V1095D, V1095R, M1096C, S1097D, K1098C, K1098V, K1098W, S1099P, S1099Q, S1100D, S1100G, S1100H, S1100M, S1100Q, Y11061, K1111R, K1111V, R1113E, R11131, R1113K, R1113N, R1113T, R1118F, R1118M, R1118Q, F1119A, F1119N, F1119W, S1120D, D1124T, T1125D, T1125E, T1125H, T1125M, T1125W, 11126A, 11126C, 11126E, T1129H, T11291, T1129V, D1131L, D1131S, E1133T, K11341, T1135Q, T1135R, T1135V, E1137L, E1137N, E1137Q, M1138D, M1138G, T1139Q, T1139V, T1139Y, 11141C, 11141P, N1142A, W1143F, D1145F, G1146K, R1150N, Q1151C, D1155M, 11158A, V1159D, F1163C, F1163N, E1164P, 11165A, L1179D, L1179E, L1179N, R1182K, R1182N, R1182Q, R1182Y, D1183Y, Y1184W, R1186M, L1187M, L1192K, L1192Y, E1194C, E1194F, E1194S, N1195S, N1195Y, N1196A, 11197G, 11197W, A1204C, A1204D, A1204F, A1204M, A1204N, A1204Q, D1206C, D1206F, D12061, A1207Q, L1208A, L1208Q, D1211C, D1211S, D1211T, A1212C, A1212M, A1212S, A1212T, A1212V, N1215G, E1227T, 11228H, 11228Y, K1229T, Q1230C, Q1230G, Q1230T, E1233W, E1233Y, N1234A, N1234E, N1234F, N12341, W1235E, W1235Y, K1236F, K1236P, E1237A, E1237F, E1237G, E12371, G1239H, G1239K, G1239L, G1239Q, G1239T, K1240Y, F1241P, S1242E, S1242Q, D12441, K1245G, K1245V, S1249E, S1249Q, S1249W, N1250A, N1250G, N1250L, N1250P, D1252L, F1254T, D1255M, F1256R, F1256W, K1260F, K1260G, K1260H, R1261M, R1261N, R1261V, R1261W, R1261Y, Y1262M, L1263A, L1263E, L1263N, L1263Q, or combinations thereof, provided that the variant ERCAS12A provides an improvement in CRISPR/ERCAS12A associated nuclease activity.

In one aspect, the ERCAS12A variant comprises at least one variant amino acid selected from amino acid positions Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, 135L/S469K, 135L/T826F, 140Q/S469K, 140Q/T826F, S469K/T826F, S469K/K516P, Y513G/F840L, Y513G/Y832R, 135L/S469K/T826F, 140Q/S469K/T826F, S469K/K516P/T826F, S469K/T826F/F840L, D303R/S469K/T826F/F840L, F465R/S469K/T826F/F840L, D303R/F465R/S469K/T826F/F840L, D303R/S469K/K516P/T826F/F840L, or combinations thereof.

Another embodiment described herein is a CRISPR ribonucleoprotein complex is provided. The CRISPR ribonucleoprotein complex includes a guide RNA and a CRISPR-associated protein. The CRISPR-associated protein includes a polypeptide encoding a variant of ERCAS12A comprising at least one variant amino acid selected from amino acid positions Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, 135L/S469K, 135L/T826F, 140Q/S469K, 140Q/T826F, S469K/T826F, S469K/K516P, Y513G/F840L, Y513G/Y832R, 135L/S469K/T826F, 140Q/S469K/T826F, S469K/K516P/T826F, S469K/T826F/F840L, D303R/S469K/T826F/F840L, F465R/S469K/T826F/F840L, D303R/F465R/S469K/T826F/F840L, D303R/S469K/K516P/T826F/F840L, or combinations thereof, provided that the variant ERCAS12A provides an improvement in CRISPR/ERCAS12A-associated nuclease activity.

Another embodiment described herein is a method of increasing efficiency of gene editing in a cell with a CRISPR ribonucleoprotein complex is provided. The method includes a step of contacting a cell with the CRISPR ribonucleoprotein complex that includes a guide RNA and a CRISPR-associated protein. The CRISPR-associated protein includes a polypeptide encoding a variant of ERCAS12A comprising at least one variant amino acid selected from amino acid positions Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, 135L/S469K, 135L/T826F, 140Q/S469K, 140Q/T826F, S469K/T826F, S469K/K516P, Y513G/F840L, Y513G/Y832R, 135L/S469K/T826F, 140Q/S469K/T826F, S469K/K516P/T826F, S469K/T826F/F840L, D303R/S469K/T826F/F840L, F465R/S469K/T826F/F840L, D303R/F465R/S469K/T826F/F840L, D303R/S469K/K516P/T826F/F840L, or combinations thereof, provided that the variant ERCAS12A provides an improvement in CRISPR/ERCAS12A-associated nuclease activity.

Another embodiment described herein is a kit including a guide RNA and a CRISPR-associated protein comprising a polypeptide encoding a variant of ERCAS12A is provided. The variant of ERCAS12A comprising at least one variant amino acid selected from amino acid positions Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, 135L/S469K, 135L/T826F, 140Q/S469K, 140Q/T826F, S469K/T826F, S469K/K516P, Y513G/F840L, Y513G/Y832R, 135L/S469K/T826F, 140Q/S469K/T826F, S469K/K516P/T826F, S469K/T826F/F840L, D303R/S469K/T826F/F840L, F465R/S469K/T826F/F840L, D303R/F465R/S469K/T826F/F840L, D303R/S469K/K516P/T826F/F840L, or combinations thereof, provided that the variant ERCAS12A provides an improvement in CRISPR/ERCAS12A-associated nuclease activity.

Another embodiment described herein is a nucleic acid encoding a CRISPR-associated protein comprising a polypeptide encoding a variant of ERCAS12A is provided. The variant of ERCAS12A comprises at least one variant amino acid selected from amino acid positions Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, 135L/S469K, 135L/T826F, 140Q/S469K, 140Q/T826F, S469K/T826F, S469K/K516P, Y513G/F840L, Y513G/Y832R, 135L/S469K/T826F, 140Q/S469K/T826F, S469K/K516P/T826F, S469K/T826F/F840L, D303R/S469K/T826F/F840L, F465R/S469K/T826F/F840L, D303R/F465R/S469K/T826F/F840L, D303R/S469K/K516P/T826F/F840L, or combinations thereof, provided that the variant ERCAS12A provides an improvement in CRISPR/ERCAS12A-associated nuclease activity.

Another embodiment described herein is a polynucleotide sequence encoding a ERCAS12A variant. The polynucleotide sequence comprises a nucleotide sequence having at least 90-99% identity of any one of the odd numbered sequences of SEQ ID NO: 3-209 (e.g., SEQ ID NO: 3, 5, 7 . . . 207, or 209). In one aspect, polynucleotide sequence comprises any one of the odd numbered sequences of SEQ ID NO: 3-209 (e.g., SEQ ID NO: 3, 5, 7 . . . 207, or 209).

Another embodiment described herein is CAS endonuclease system comprising an expression cassette encoding a polynucleotide sequence encoding a ERCAS12A polypeptide is provided. The polynucleotide sequence comprises a nucleotide sequence having at least 90-99% identity of any one of the odd numbered sequences of SEQ ID NO: 3-209 (e.g., 3, 5, 7 . . . 207, or 209). In one aspect, polynucleotide sequence comprises any one of the odd numbered sequences of SEQ ID NO: 3-209 (e.g., 3, 5, 7 . . . 207, or 209).

Another embodiment described herein is a CAS endonuclease system comprising an amino acid sequence encoding a ERCAS12A polypeptide. The amino acid sequence includes a polypeptide comprising a ERCAS12A variant selected from a polypeptide having one or more amino acid substitutions presented in Table 1 relative to wild type ERCAS12A (SEQ ID NO: 2), wherein the ERCAS12A variant displays improved genome editing relative to wild type ERCAS12A (SEQ ID NO: 2).

Another embodiment described herein is a method of performing genome editing in eukaryotic cells. The method includes the step of introducing an CAS endonuclease system into the eukaryotic cell, said CAS endonuclease system comprising an expression cassette encoding a polynucleotide sequence encoding a ERCAS12A polypeptide, comprising a ERCAS12A variant selected from a polypeptide having one or more amino acid substitutions presented in Table 1 relative to wild type ERCAS12A (SEQ ID NO: 2), wherein the ERCAS12A variant displays improved genome editing relative to wild type ERCAS12A (SEQ ID NO: 2).

Another embodiment described herein is a method of performing genome editing in a eukaryotic cell. The method includes the step of introducing an CAS endonuclease system into the eukaryotic cell, said CAS endonuclease system comprising an amino acid sequence encoding a ERCAS12A polypeptide comprising a ERCAS12A variant selected from a polypeptide having one or more amino acid substitutions presented in Table 1 relative to wild type ERCAS12A (SEQ ID NO: 2), wherein the ERCAS12A variant displays improved genome editing relative to wild type ERCAS12A (SEQ ID NO: 2) Another embodiment described herein is an CRISPR-associated protein comprising a fusion polypeptide. The fusion polypeptide includes an ERCAS12A open reading frame, a nuclear localization signal, optionally an amino acid linker and optionally an affinity tag.

Another embodiment is a polypeptide encoded by a nucleotide sequence described herein. In one aspect, the polypeptide has at least 85% to 99% identity to the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210). In another aspect, the polypeptide is selected from the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210).

Another embodiment described herein is a process for manufacturing one or more of the nucleotide sequence described herein or a polypeptide encoded by the nucleotide sequence described herein, the process comprising: transforming or transfecting a cell with a nucleic acid comprising a nucleotide sequence described herein; growing the cells; optionally isolating additional quantities of a nucleotide sequence described herein; inducing expression of a polypeptide encoded by a nucleotide sequence of described herein; isolating the polypeptide encoded by a nucleotide described herein.

Another embodiment described herein is a means for manufacturing one or more of the nucleotide sequences described herein or a polypeptide encoded by a nucleotide sequence described herein, the process comprising: transforming or transfecting a cell with a nucleic acid comprising a nucleotide sequence described herein; growing the cells; optionally isolating additional quantities of a nucleotide sequence described herein; inducing expression of a polypeptide encoded by a nucleotide sequence of described herein; isolating the polypeptide encoded by a nucleotide described herein.

Another embodiment described herein is a nucleotide sequence or a polypeptide encoded by the nucleotide sequence produced by the methods or the means described herein.

Another embodiment described herein is the use of an effective amount of a polypeptide encoded by one or more of the odd numbered sequences of SEQ ID NO: 3-209 (e.g., 3, 5, 7 . . . 207, or 209).

Another embodiment described herein is a research tool comprising a polypeptide encoded by a nucleotide sequence described herein.

Another embodiment described herein is a reagent comprising a polypeptide encoded by a nucleotide sequence described herein.

The polynucleotides described herein include variants that have substitutions, deletions, and/or additions that can involve one or more nucleotides. The variants can be altered in coding regions, non-coding regions, or both. Alterations in the coding regions can produce conservative or non-conservative amino acid substitutions, deletions, or additions. Especially preferred among these are silent substitutions, additions, and deletions, which do not alter the properties and activities of the binding.

Further embodiments described herein include nucleic acid molecules comprising polynucleotides having nucleotide sequences at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, and more preferably at least about 90-99% or 100% identical to (a) nucleotide sequences, or degenerate, homologous, or codon-optimized variants thereof, encoding polypeptides having the amino acid sequences in the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210); (b) nucleotide sequences, or degenerate, homologous, or codon-optimized variants thereof, encoding polypeptides having the amino acid sequences in the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210); and (c) nucleotide sequences capable of hybridizing to the complement of any of the nucleotide sequences in (a) or (b) above and capable of expressing functional polypeptides of amino acid sequences the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210).

By a polynucleotide having a nucleotide sequence at least, for example, 90-99% “identical” to a reference nucleotide sequence encoding the odd numbered sequences of SEQ ID NO: 3-209 (e.g., 3, 5, 7 . . . 207, or 209) is intended that the nucleotide sequence of the polynucleotide be identical to the reference sequence except that the polynucleotide sequence can include up to about 10 to 1 point mutations, additions, or deletions per each 100 nucleotides of the reference nucleotide sequence encoding the odd numbered sequences of SEQ ID NO: 3-209 (e.g., 3, 5, 7 . . . 207, or 209).

In other words, to obtain a polynucleotide having a nucleotide sequence about at least 90-99% identical to a reference nucleotide sequence, up to 10% of the nucleotides in the reference sequence can be deleted, added, or substituted, with another nucleotide, or a number of nucleotides up to 10% of the total nucleotides in the reference sequence can be inserted into the reference sequence. These mutations of the reference sequence can occur at the 5′- or 3′-terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence. The same is applicable to polypeptide sequences at least about 90-99% identical to a reference polypeptide sequence.

As noted above, two or more polynucleotide sequences can be compared by determining their percent identity. Two or more amino acid sequences likewise can be compared by determining their percent identity. The percent identity of two sequences, whether nucleic acid or peptide sequences, is generally described as the number of exact matches between two aligned sequences divided by the length of the shorter sequence and multiplied by 100. An approximate alignment for nucleic acid sequences is provided by the local homology algorithm of Smith and Waterman, Advances in Applied Mathematics 2: 4 82-489 (1981). This algorithm can be extended to use with peptide sequences using the scoring matrix developed by Dayhoff, Atlas of Protein Sequences and Structure, M. O. Dayhoff ed., 5 suppl. 3: 353-358, National Biomedical Research Foundation, Washington, D.C., USA, and normalized by Gribskov, Nucl. Acids Res. 14(6): 6745-6763 (1986).

For example, due to the degeneracy of the genetic code, one having ordinary skill in the art will recognize that a large number of the nucleic acid molecules having a sequence at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acid sequence shown in the odd numbered sequences of SEQ ID NO: 3-209 (e.g., 3, 5, 7 . . . 207, or 209), or degenerate, homologous, or codon-optimized variants thereof, will encode the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210).

The polynucleotides described herein include those encoding mutations, variations, substitutions, additions, deletions, and particular examples of the polypeptides described herein. For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie, J. U. et al., “Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions,” Science 247: 1306-1310 (1990), wherein the authors indicate that proteins are surprisingly tolerant of amino acid substitutions.

Thus, fragments, derivatives, or analogs of the polypeptides of the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210) can be (i) ones in which one or more of the amino acid residues (e.g., 1, 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 residues, or even more) are substituted with a conserved or non-conserved amino acid residue (preferably a conserved amino acid residue). Such substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) ones in which one or more of the amino acid residues includes a substituent group (e.g., 1, 2, 3, 4, 5, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 residues or even more), or (iii) ones in which the mature polypeptide is fused with another polypeptide or compound, such as a compound to increase the half-life of the polypeptide (for example, polyethylene glycol), or (iv) ones in which the additional amino acids are fused to the mature polypeptide, such as an IgG Fc fusion region peptide or leader or secretory sequence or a sequence which is employed for purification of the mature polypeptide or a proprotein sequence. Such fragments, derivatives, and analogs are deemed to be within the scope of those skilled in the art from the teachings herein.

In addition, fragments, derivatives, or analogs of the polypeptides of the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210) can be substituted with one or more conserved or non-conserved amino acid residue (preferably a conserved amino acid residue). In some cases, these polypeptides, fragments, derivatives, or analogs thereof will have a polypeptide sequence at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the polypeptide sequence shown in the even numbered sequences of SEQ ID NO: 4-210 (e.g., SEQ ID NO: 4, 6, 8 . . . 206, or 210) and will comprise functional or non-functional proteins or enzymes. Similarly, additions or deletions to the polypeptides can be made either at the N- or C-termini or within non-conserved regions of the polypeptide (which are assumed to be non-critical because they have not been photogenically conserved).

As described herein, in many cases the amino acid substitutions, mutations, additions, or deletions are preferably of a minor nature, such as conservative amino acid substitutions that do not significantly affect the folding or activity of the protein or additions or deletions to the N- or C-termini. Of course, the number of amino acid substitutions, additions, or deletions a skilled artisan would make depends on many factors, including those described herein. Generally, the number of substitutions, additions, or deletions for any given polypeptide will not be more than about 100, 90, 80, 70, 60, 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 5, 6, 4, 3, 2, or 1.

Another embodiment described herein is a polynucleotide vector comprising one or more nucleotide sequences described herein.

Another embodiment described herein is a cell comprising one or more nucleotide sequences described herein or a polynucleotide vector described herein.

It will be apparent to one of ordinary skill in the relevant art that suitable modifications and adaptations to the compositions, formulations, methods, processes, apparata, assemblies, and applications described herein can be made without departing from the scope of any embodiments or aspects thereof. The compositions, apparata, assemblies, and methods provided are exemplary and are not intended to limit the scope of any of the disclosed embodiments. All the various embodiments, aspects, and options disclosed herein can be combined in any variations or iterations. The scope of the compositions, formulations, methods, apparata, assemblies, and processes described herein include all actual or potential combinations of embodiments, aspects, options, examples, and preferences described herein. The compositions, formulations, apparata, assemblies, or methods described herein may omit any component or step, substitute any component or step disclosed herein, or include any component or step disclosed elsewhere herein. The ratios of the mass of any component of any of the compositions or formulations disclosed herein to the mass of any other component in the formulation or to the total mass of the other components in the formulation are hereby disclosed as if they were expressly disclosed. Should the meaning of any terms in any of the patents or publications incorporated by reference conflict with the meaning of the terms used in this disclosure, the meanings of the terms or phrases in this disclosure are controlling. All patents and publications cited herein are incorporated by reference herein for the specific teachings thereof.

Various embodiments and aspects of the inventions described herein are summarized by the following clauses:

- Clause 1. An isolated polypeptide comprising an Eubacterium rectale Cas12a (ERCAS12A) mutant having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant polypeptide provides an improvement in nuclease activity as compared to the wild-type ERCAS12A polypeptide.
- Clause 2. The isolated polypeptide of clause 1, wherein the ERCAS12A mutant has 98% to 99% identity to a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 3. The isolated polypeptide of clause 1 or 2, wherein the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 4. The isolated polypeptide of any one of clauses 1-3, wherein the ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 5. The isolated polypeptide of any one of clauses 1-4, wherein the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 6. The isolated polypeptide of any one of clauses 1-5, wherein the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291 N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 7. The isolated polypeptide of any one of clauses 1-6, wherein the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 8. An isolated polynucleotide sequence encoding an Eubacterium rectale Cas12a (ERCAS12A) mutant having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide.
- Clause 9. The isolated polynucleotide sequence of clause 8, wherein the encoded ERCAS12A mutant has 98% to 99% identity to a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 10. The isolated polynucleotide sequence of clause 8 or 9, wherein the encoded ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 11. The isolated polynucleotide sequence of any one of clauses 8-10, wherein the ERCAS12A mutant polynucleotide sequence has 95% to 99% identity to a nucleotide sequence selected from the odd numbered sequences of SEQ ID NO: 3-209.
- Clause 12. The isolated polynucleotide sequence of any one of clauses 8-11, wherein the ERCAS12A mutant polynucleotide has a nucleotide sequence selected from the odd numbered sequences of SEQ ID NO: 3-209.
- Clause 13. The isolated polynucleotide sequence of any one of clauses 8-12, wherein the encoded ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 14. The isolated polynucleotide sequence of any one of clauses 8-13, wherein the encoded ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 15. The isolated polynucleotide sequence of any one of clauses 8-14, wherein encoded ERCAS12A mutant polynucleotide is selected from is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 16. The isolated polynucleotide sequence of any one of clauses 8-15, wherein the ERCAS12A mutant poly nucleotide is selected from Q18V (SEQ ID NO: 3); 135L (SEQ ID NO: 5); G39Q (SEQ ID NO: 7); G39K (SEQ ID NO: 9); 140Q (SEQ ID NO: 11); D56R (SEQ ID NO: 13); D56P (SEQ ID NO: 15); D59R (SEQ ID NO: 17); D59A (SEQ ID NO: 19); S158G (SEQ ID NO: 21); K165Y (SEQ ID NO: 23); S181V (SEQ ID NO: 25); A290K (SEQ ID NO: 27); D291N (SEQ ID NO: 29); V296R (SEQ ID NO: 31); P297V (SEQ ID NO: 33); P297R (SEQ ID NO: 35); P297F (SEQ ID NO: 37); D303R (SEQ ID NO: 39); D303C (SEQ ID NO: 41); E305K (SEQ ID NO: 43); Y307K (SEQ ID NO: 45); Q353R (SEQ ID NO: 47); L444W (SEQ ID NO: 49); F465R (SEQ ID NO: 51); S469K (SEQ ID NO: 53); V470K (SEQ ID NO: 55); 1498K (SEQ ID NO: 57); Y507W (SEQ ID NO: 59); Y507S (SEQ ID NO: 61); Y507R (SEQ ID NO: 63); Q510Y (SEQ ID NO: 65); Q510V (SEQ ID NO: 67); Q51OK (SEQ ID NO: 69); P512V (SEQ ID NO: 71); P512G (SEQ ID NO: 73); Y513S (SEQ ID NO: 75); Y513R (SEQ ID NO: 77); Y513L (SEQ ID NO: 79); Y513G (SEQ ID NO: 81); S514R (SEQ ID NO: 83); S514P (SEQ ID NO: 85); T515P (SEQ ID NO: 87); K516R (SEQ ID NO: 89); K516P (SEQ ID NO: 91); K517P (SEQ ID NO: 93); 1518P (SEQ ID NO: 95); L520M (SEQ ID NO: 97); F522A (SEQ ID NO: 99); A541V (SEQ ID NO: 101); G553M (SEQ ID NO: 103); N556R (SEQ ID NO: 105); N556E (SEQ ID NO: 107); D562V (SEQ ID NO: 109); D562K (SEQ ID NO: 111); D5621 (SEQ ID NO: 113); N583V (SEQ ID NO: 115); L585K (SEQ ID NO: 117); F653R (SEQ ID NO: 119); K712R (SEQ ID NO: 121); N716R (SEQ ID NO: 123); E731P (SEQ ID NO: 125); N732T (SEQ ID NO: 127); A744T (SEQ ID NO: 129); A744P (SEQ ID NO: 131); A744N (SEQ ID NO: 132); F748A (SEQ ID NO: 135); S751A (SEQ ID NO: 137); K754G (SEQ ID NO: 139); 1757W (SEQ ID NO: 141); 1757K (SEQ ID NO: 143); E771R (SEQ ID NO: 145); N787A (SEQ ID NO: 147); A825R (SEQ ID NO: 149); T826K (SEQ ID NO: 151); T826F (SEQ ID NO: 153); Y832R (SEQ ID NO: 155); Y836Q (SEQ ID NO: 157); Y836K (SEQ ID NO: 159); Y839L (SEQ ID NO: 161); F840L (SEQ ID NO: 163); Y907V (SEQ ID NO: 165); Y9071 (SEQ ID NO: 167); Q916Y (SEQ ID NO: 169); E935R (SEQ ID NO: 171); 1936R (SEQ ID NO: 173); K974G (SEQ ID NO: 175); K1019V (SEQ ID NO: 177); 135L/S469K (SEQ ID NO: 179); 135L/T826F (SEQ ID NO: 181); 140Q/S469K (SEQ ID NO: 183); 140Q/T826F (SEQ ID NO: 185); S469K/K516P (SEQ ID NO: 187); S469K/T826F (SEQ ID NO: 189); Y513G/Y832R (SEQ ID NO: 191); Y513G/F840L (SEQ ID NO: 193); 135L/S469K/T826F (SEQ ID NO: 195); 140Q/S469K/T826F (SEQ ID NO: 197); S469K/K516P/T826F (SEQ ID NO: 199); S469K/T826F/F840L (SEQ ID NO: 201); D303R/S469K/T826F/F840L (SEQ ID NO: 203); F465R/S469K/T826F/F840L (SEQ ID NO: 205); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 207); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 209).
- Clause 17. The isolated polynucleotide sequence of any one of clauses 8-16, wherein the ERCAS12A mutant polynucleotide is selected from is selected from: D303R/S469K/T826F/F840L (SEQ ID NO: 203); F465R/S469K/T826F/F840L (SEQ ID NO: 205); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 207); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 209).
- Clause 18. A vector or plasmid comprising the polynucleotide sequence of any one of clauses 8-17.
- Clause 19. A cell comprising a nucleic acid comprising one or more of the polynucleotide sequences of any one of clauses 8-18.
- Clause 20. An isolated CRISPR ribonucleoprotein complex comprising a guide RNA and a ERCAS12A mutant polypeptide having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide.
- Clause 21. The isolated CRISPR ribonucleoprotein complex of clause 20, wherein the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 22. The isolated CRISPR ribonucleoprotein complex of clause 20 or 21, wherein the ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 23. The isolated CRISPR ribonucleoprotein complex of any one of clauses 20-22, wherein the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 24. The isolated CRISPR ribonucleoprotein complex of any one of clauses 20-23, wherein the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 25. The isolated CRISPR ribonucleoprotein complex of any one of clauses 20-24, wherein the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 26. An isolated CRISPR/Cpf1 endonuclease system comprising a crRNA and a ERCAS12A mutant polypeptide having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant polypeptide provides improved genome editing as compared to the wild-type ERCAS12A polypeptide.
- Clause 27. The isolated CRISPR/Cpf1 endonuclease system of clause 26, wherein the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 28. The isolated CRISPR/Cpf1 endonuclease system of clause 26 or 27, wherein the ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 29. The isolated CRISPR/Cpf1 endonuclease system of any one of clauses 26-28, wherein the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 30. The isolated CRISPR/Cpf1 endonuclease system of any one of clauses 26-29, wherein the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 31. The isolated CRISPR/Cpf1 endonuclease system of any one of clauses 26-30, wherein the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 32. A method for performing gene editing comprising:
  - contacting a candidate editing target site locus with a CRISPR ribonucleoprotein complex comprising a crRNA and an ERCAS12A mutant polypeptide comprising having one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide.
- Clause 33. The method of clause 32, wherein the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 34. The method of clause 32 or 33, wherein the ERCAS12A mutant comprises ERCAS12A mutant comprises one or more amino acid substitution at positions 18, 35, 39, 39, 40, 56, 56, 59, 59, 158, 165, 181, 290, 291, 296, 297, 297, 297, 303, 303, 305, 307, 353, 444, 465, 469, 470, 498, 507, 507, 507, 510, 510, 510, 512, 512, 513, 513, 513, 513, 514, 514, 515, 516, 516, 517, 518, 520, 522, 541, 553, 556, 556, 562, 562, 562, 583, 585, 653, 712, 716, 731, 732, 744, 744, 744, 748, 751, 754, 757, 757, 771, 787, 825, 826, 826, 832, 836, 836, 839, 840, 907, 907, 916, 935, 936, 974, or 1019 of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 35. The method of any one of clauses 32-34, wherein the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 36. The method of any one of clauses 32-35, wherein the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 37. The method of any one of clauses 32-36, wherein the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 38. A kit comprising a CRISPR ribonucleoprotein complex comprising a guide RNA and an ERCAS12A mutant polypeptide comprising one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide.
- Clause 39. The kit of clause 38, wherein the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 40. The kit of clause 38 or 39, wherein the ERCAS12A mutant comprises wherein the ERCAS12A mutant comprises one or more amino acid substitutions selected from Q18V, 135L, G39K, G39Q, 140Q, D56P, D56R, D59A, D59R, S158G, K165Y, S181V, A290K, D291N, V296R, P297F, P297R, P297V, D303C, D303R, E305K, Y307K, Q353R, L444W, F465R, S469K, V470K, 1498K, Y507R, Y507S, Y507W, Q510K, Q510V, Q510Y, P512G, P512V, Y513G, Y513L, Y513R, Y513S, S514P, S514R, T515P, K516P, K516R, K517P, 1518P, L520M, F522A, A541V, G553M, N556E, N556R, D5621, D562K, D562V, N583V, L585K, F653R, K712R, N716R, E731P, N732T, A744N, A744P, A744T, F748A, S751A, K754G, 1757K, 1757W, E771R, N787A, A825R, T826F, T826K, Y832R, Y836K, Y836Q, Y839L, F840L, Y9071, Y907V, Q916Y, E935R, 1936R, K974G, K1019V, or combinations thereof of the wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2.
- Clause 41. The kit of any one of clauses 38-40, wherein the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 42. The kit of any one of clauses 38-41, wherein the ERCAS12A mutant is selected from D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).
- Clause 43. Use of a ERCAS12A mutant polypeptide for improving gene editing, wherein the ERCAS12A mutant one or more amino acid substitutions introduced into a wild-type ERCAS12A polypeptide sequence of SEQ ID NO: 2, where the ERCAS12A mutant provides improved gene editing as compared to the wild-type ERCAS12A polypeptide.
- Clause 44. The use of clause 43, wherein the ERCAS12A mutant has a polypeptide sequence selected from the even numbered sequences of SEQ ID NO: 4-210.
- Clause 45. The use of clause 43 or 44, wherein the ERCAS12A mutant is selected from: Q18V (SEQ ID NO: 4); 135L (SEQ ID NO: 6); G39Q (SEQ ID NO: 8); G39K (SEQ ID NO: 10); 140Q (SEQ ID NO: 12); D56R (SEQ ID NO: 14); D56P (SEQ ID NO: 16); D59R (SEQ ID NO: 18); D59A (SEQ ID NO: 20); S158G (SEQ ID NO: 22); K165Y (SEQ ID NO: 24); S181V (SEQ ID NO: 26); A290K (SEQ ID NO: 28); D291 N (SEQ ID NO: 30); V296R (SEQ ID NO: 32); P297V (SEQ ID NO: 34); P297R (SEQ ID NO: 36); P297F (SEQ ID NO: 38); D303R (SEQ ID NO: 40); D303C (SEQ ID NO: 42); E305K (SEQ ID NO: 44); Y307K (SEQ ID NO: 46); Q353R (SEQ ID NO: 48); L444W (SEQ ID NO: 50); F465R (SEQ ID NO: 52); S469K (SEQ ID NO: 54); V470K (SEQ ID NO: 56); 1498K (SEQ ID NO: 58); Y507W (SEQ ID NO: 60); Y507S (SEQ ID NO: 62); Y507R (SEQ ID NO: 64); Q510Y (SEQ ID NO: 66); Q510V (SEQ ID NO: 68); Q510K (SEQ ID NO: 70); P512V (SEQ ID NO: 72); P512G (SEQ ID NO: 74); Y513S (SEQ ID NO: 76); Y513R (SEQ ID NO: 78); Y513L (SEQ ID NO: 80); Y513G (SEQ ID NO: 82); S514R (SEQ ID NO: 84); S514P (SEQ ID NO: 86); T515P (SEQ ID NO: 88); K516R (SEQ ID NO: 90); K516P (SEQ ID NO: 92); K517P (SEQ ID NO: 94); 1518P (SEQ ID NO: 96); L520M (SEQ ID NO: 98); F522A (SEQ ID NO: 100); A541V (SEQ ID NO: 102); G553M (SEQ ID NO: 104); N556R (SEQ ID NO: 106); N556E (SEQ ID NO: 108); D562V (SEQ ID NO: 110); D562K (SEQ ID NO: 112); D5621 (SEQ ID NO: 114); N583V (SEQ ID NO: 116); L585K (SEQ ID NO: 118); F653R (SEQ ID NO: 120); K712R (SEQ ID NO: 122); N716R (SEQ ID NO: 124); E731P (SEQ ID NO: 126); N732T (SEQ ID NO: 128); A744T (SEQ ID NO: 130); A744P (SEQ ID NO: 132); A744N (SEQ ID NO: 134); F748A (SEQ ID NO: 136); S751A (SEQ ID NO: 138); K754G (SEQ ID NO: 140); 1757W (SEQ ID NO: 142); 1757K (SEQ ID NO: 144); E771R (SEQ ID NO: 146); N787A (SEQ ID NO: 148); A825R (SEQ ID NO: 150); T826K (SEQ ID NO: 152); T826F (SEQ ID NO: 154); Y832R (SEQ ID NO: 156); Y836Q (SEQ ID NO: 158); Y836K (SEQ ID NO: 160); Y839L (SEQ ID NO: 162); F840L (SEQ ID NO: 164); Y907V (SEQ ID NO: 166); Y9071 (SEQ ID NO: 168); Q916Y (SEQ ID NO: 170); E935R (SEQ ID NO: 172); 1936R (SEQ ID NO: 174); K974G (SEQ ID NO: 176); K1019V (SEQ ID NO: 178); 135L/S469K (SEQ ID NO: 180); 135L/T826F (SEQ ID NO: 182); 140Q/S469K (SEQ ID NO: 184); 140Q/T826F (SEQ ID NO: 186); S469K/K516P (SEQ ID NO: 188); S469K/T826F (SEQ ID NO: 190); Y513G/Y832R (SEQ ID NO: 192); Y513G/F840L (SEQ ID NO: 194); 135L/S469K/T826F (SEQ ID NO: 196); 140Q/S469K/T826F (SEQ ID NO: 198); S469K/K516P/T826F (SEQ ID NO: 200); S469K/T826F/F840L (SEQ ID NO: 202); D303R/S469K/T826F/F840L (SEQ ID NO: 204); F465R/S469K/T826F/F840L (SEQ ID NO: 206); D303R/F465R/S469K/T826F/F840L (SEQ ID NO: 208); or D303R/S469K/K516P/T826F/F840L (SEQ ID NO: 210).

EXAMPLES
Example 1

High-Throughput Measurement of DNA Cleavage Activity of ERCAS12A Variants at TTTT PAM Sites in E. coli.

The following Example demonstrates the ability of the invention to increase the efficiency of gene editing as compared to ERCAS12A (variant of E. rectalis Cas12a). The wild type nucleotide sequence of ERCAS12A (SEQ ID NO: 1) and amino acid sequence (SEQ ID NO: 2) are shown in Table 1.

TABLE 1

ERCAS12A Polynucleotide and Polypeptide

Sequences

Wild Type ERCAS12A Nucleotide Sequence

(SEQ ID NO: 1)

ATGAATAACGGCACCAACAACTTCCAGAACTTTATTGGTATTAGC

AGCCTGCAGAAAACCCTGCGTAATGCACTGATTCCGACCGAAACC

ACACAGCAGTTTATTGTGAAAAACGGCATCATCAAAGAAGATGAA

CTTCGCGGAGAAAATCGCCAGATTCTGAAAGATATCATGGATGAT

TATTATCGCGGTTTTATCAGCGAAACCCTGAGCAGCATTGATGAT

ATTGATTGGACCAGCCTGTTCGAGAAAATGGAAATCCAGCTGAAA

AATGGCGATAACAAAGACACCCTGATTAAAGAACAGGCCGAAAAA

CGTAAAGCCATCTACAAAAAATTCGCCGATGATGACCGCTTTAAA

AACATGTTCAGCGCAAAACTGATTAGCGATATTCTGCCGGAATTT

GTGATCCACAATAATAACTATAGCGCCAGCGAGAAAGAAGAGAAA

ACCCAGGTTATTAAACTGTTTAGCCGTTTTGCGACCAGCTTCAAA

GATTACTTCAAAAATCGTGCCAACTGCTTCAGCGCAGATGATATT

AGCAGTAGCAGCTGTCATCGTATTGTGAATGATAACGCCGAAATC

TTTTTTAGCAATGCCCTGGTTTATCGTCGCATCGTTAAAAATCTG

AGCAACGACGATATCAACAAAATCAGCGGTGATATGAAAGACAGC

CTGAAAGAAATGAGCCTGGAAAAAATCTACAGCTACGAAAAATAT

GGCGAGTTCATTACCCAAGAAGGCATCAGCTTTTATAACGATATT

TGCGGCAAAGTGAACAGCTTTATGAATCTGTATTGCCAGAAAAAC

AAAGAGAACAAAAACCTGTACAAACTGCGCAAACTGCATAAGCAG

ATTCTGTGTATTGCAGATACCAGCTATGAAGTGCCGTATAAATTC

GAAAGTGATGAAGAAGTGTACCAGAGCGTTAATGGTTTTCTGGAT

AACATCAGCAGCAAACACATTGTTGAACGTCTGCGTAAAATTGGC

GACAACTATAATGGCTATAACCTGGACAAAATTTACATCGTGAGC

AAGTTTTATGAACTGGTGAGCCAGAAAACATATCGTGATTGGGAA

ACAATTAATACCGCCTTGGAGATCCACTACAATAACATTCTGCCT

GGTAATGGTAAAAGCAAAGCCGACAAAGTTAAAAAGGCCGTGAAA

AATGATCTGCAGAAATCCATTACCGAAATCAACGAACTGGTCAGC

AACTATAAACTGTGTCCGGATGATAATATCAAGGCCGAAACCTAT

ATTCACGAAATCTCCCATATCCTGAACAACTTTGAAGCACAAGAG

CTGAAATATAACCCGGAAATTCATCTGGTTGAAAGCGAACTGAAA

GCCAGTGAACTGAAAAACGTTCTGGATGTGATTATGAACGCCTTT

CATTGGTGTAGCGTTTTTATGACCGAAGAACTGGTGGATAAGGAT

AACAACTTTTATGCCGAACTGGAAGAGATCTATGATGAAATTTAT

CCGGTGATCAGCCTGTATAATCTGGTGCGTAATTATGTGACCCAG

AAACCGTATAGCACCAAAAAAATCAAACTGAACTTTGGCATTCCG

ACACTGGCAGATGGTTGGAGCAAAAGTAAAGAATATAGCAACAAC

GCCATTATCCTGATGCGCGATAATCTGTACTATCTGGGTATCTTT

AACGCCAAAAACAAACCGGACAAAAAGATCATTGAAGGCAATACC

AGCGAAAACAAAGGCGACTATAAAAAGATGATCTACAATCTGCTG

CCTGGTCCGAATAAAATGATTCCGAAAGTTTTCCTGAGCAGTAAA

ACCGGTGTGGAAACATATAAACCGAGCGCATATATTCTGGAAGGC

TACAAACAGAACAAGCATCTGAAAAGCAGCAAAGACTTTGATATC

ACCTTTTGCCGTGATCTGATCGACTATTTCAAAAACTGCATTGCC

ATTCATCCGGAATGGAAAAACTTTGGCTTTGATTTTAGCGATACC

AGCACCTATGAAGATATCAGCGGTTTTTATCGTGAAGTTGAACTG

CAGGGCTATAAAATCGATTGGACCTATATTAGCGAGAAGGATATT

GATCTGCTGCAAGAAAAAGGTCAGCTGTACCTGTTTCAGATCTAC

AACAAAGACTTCAGCAAAAAAAGCACCGGCAATGATAATCTGCAT

ACCATGTATCTGAAGAACCTGTTTAGCGAAGAGAACCTGAAAGAC

ATTGTGCTGAAACTGAATGGTGAAGCCGAAATTTTCTTTCGTAAG

AGCAGCATCAAAAACCCGATCATCCATAAAAAAGGTAGCATTCTG

GTGAATCGTACGTATGAAGCAGAAGAAAAAGACCAGTTTGGCAAC

ATTCAGATTGTGCGTAAAACAATCCCGGAAAACATCTATCAAGAG

CTGTACAAATACTTTAACGATAAAAGCGATAAAGAACTGTCCGAT

GAAGCAGCCAAACTGAAGAATGTTGTTGGTCATCATGAAGCCGCA

ACCAACATTGTTAAAGATTATCGTTACACCTATGATAAATACTTT

CTGCACATGCCGATCACCATCAACTTTAAAGCAAATAAGACGAGC

TTCATCAACGATCGCATTCTGCAGTATATCGCCAAAGAAAAGGAT

CTGCATGTGATTGGTATTGATCGTGGTGAACGCAATCTGATTTAT

GTGAGCGTTATTGATACCTGCGGCAATATTGTGGAACAGAAAAGC

TTTAATATCGTGAACGGCTATGACTACCAGATCAAGCTGAAACAG

CAAGAAGGTGCACGTCAGATTGCACGTAAAGAATGGAAAGAAATC

GGCAAGATCAAAGAAATCAAAGAGGGTTATCTGAGCCTGGTGATT

CATGAAATTAGCAAAATGGTGATCAAATATAACGCCATCATTGCC

ATGGAAGATCTGAGTTATGGTTTCAAAAAAGGTCGCTTTAAGGTG

GAACGTCAGGTGTATCAGAAATTTGAAACGATGCTGATCAACAAA

CTGAATTATCTGGTGTTCAAGGACATCAGCATTACGGAAAATGGT

GGTCTGCTGAAAGGTTATCAGCTGACGTATATTCCGGAAAAGCTT

AAAAATGTGGGTCATCAGTGTGGCTGCATCTTTTATGTTCCGGCA

GCCTATACCAGCAAAATTGATCCGACCACCGGCTTTGTTAACATC

TTCAAATTCAAAGATCTGACCGTGGATGCCAAACGCGAATTTATC

AAAAAGTTTGATAGCATCCGCTACGACAGCGAGAAGAACTTATTT

TGTTTCACCTTCGACTACAACAACTTTATCACCCAGAATACGGTT

ATGAGCAAAAGCAGCTGGTCAGTTTATACCTATGGTGTGCGTATT

AAACGTCGCTTTGTGAATGGTCGCTTTTCAAATGAAAGCGACACC

ATCGATATCACCAAAGACATGGAAAAAACCCTGGAAATGACCGAT

ATTAACTGGCGTGATGGTCATGATCTGCGTCAGGATATTATCGAT

TATGAAATCGTGCAGCACATCTTTGAAATTTTTCGTCTGACCGTT

CAGATGCGTAATAGCCTGAGCGAGCTGGAAGATCGCGATTATGAT

CGTCTGATTAGTCCGGTTCTGAACGAAAACAACATCTTCTATGAT

AGCGCAAAAGCGGGTGATGCACTGCCGAAAGATGCCGATGCAAAT

GGTGCATATTGTATTGCACTGAAAGGCCTGTATGAAATCAAGCAG

ATTACCGAGAATTGGAAAGAGGATGGTAAATTCAGCCGTGATAAA

CTGAAAATCTCCAACAAGGATTGGTTCGATTTCATCCAGAATAAA

CGCTACCTG

Wild Type ERCAS12A Polypeptide Sequence

(SEQ ID NO: 2)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDE

LRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEKMEIQLK

NGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEF

VIHNNNYSASEKEEKTQVIKLESRFATSFKDYFKNRANCESADDI

SSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDS

LKEMSLEEIYSYEKYGEFITQEGISFYNDICGKVNSFMNLYCQKN

KENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELD

NISSKHIVERLRKIGDNYNGYNLDKIYIVSKFYESVSQKTYRDWE

TINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVS

NYKLCSDDNIKAETYIHEISHILNNFEAQELKYNPEIHLVESELK

ASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIY

PVISLYNLVRNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNN

AIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLL

PGPNKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDI

TFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYREVEL

QGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLH

TMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNPIIHKKGSIL

VNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSD

EAAKLKNVVGHHEAATNIVKDYRYTYDKYFLHMPITINFKANKTG

FINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKS

FNIVNGYDYQIKLKQQEGARQIARKEWKEIGKIKEIKEGYLSLVI

HEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINK

LNYLVFKDISITENGGLLKGYQLTYIPDKLKNVGHQCGCIFYVPA

AYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLF

CFTFDYNNFITQNTVMSKSSWSVYTYGVRIKRRFVNGRESNESDT

IDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTV

QMRNSLSELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADAN

GAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

The phenotype scores of 22,409 ERCAS12A variants with single point mutation are listed in Table 2. Scores for synonymous changes (n=849, 69% of all positions) tightly clustered around 0, and thus serving as the baseline activity of ERCAS12A (SEQ ID NO: 2) (FIG. 1A). The enrichment measure for each ERCAS12A point mutation was highly consistent across biological replicates (FIG. 11B). Overall, 2,564 of 22,409 (˜11.4%) point mutations exhibited some benefits with score greater than the maximum score (0.41) of all synonymous changes (Table 2).

Specific mutated residues that conferred some benefit are shown in the sequence of the wild type ERCAS12A Polypeptide Sequence (SEQ ID NO: 2). Mutated residues are bolded; non-bold residues were not mutated. There were 2564 individual mutations at 763 specific residues of the 1263 amino acid residues (˜60%).

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDE

LRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEKMEIQLK

NGDNKDTLIKEQAEKRKAIYKKFADDDRFKNMESAKLISDILPEF

VIHNNNYSASEKEEKTQVIKLFSRFATSFKDYFKNRANCFSADDI

SSSSCHRIVNDNAEIFFSNALVYRRIVKNLSNDDINKISGDMKDS

LKEMSLEKIYSYEKYGEFITQEGISFYNDICGKVNSFMNLYCQKN

KENKNLYKLRKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELD

NISSKHIVERLRKIGDNYNGYNLDKIYIVSKFYELVSQKTYRDWE

TINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVS

NYKLCPDDNIKAETYIHEISHILNNFEAQELKYNPEIHLVESELK

ASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIY

PVISLYNLVRNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNN

AIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLL

PGPNKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHLKSSKDFDI

TFCRDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYREVEL

QGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLH

TMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNPIIHKKGSIL

VNRTYEAEEKDQFGNIQIVRKTIPENIYQELYKYENDKSDKELSD

EAAKLKNVVGHHEAATNIVKDYRYTYDKYFLHMPITINFKANKTS

FINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKS

ENIVNGYDYQIKLKQQEGARQIARKEWKEIGKIKEIKEGYLSLVI

HEISKMVIKYNAIIAMEDLSYGEKKGRFKVERQVYQKFETMLINK

LNYLVFKDISITENGGLLKGYQLTYIPEKLKNVGHQCGCIFYVPA

AYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLF

CFTFDYNNFITQNTVMSKSSWSVYTYGVRIKRRFVNGRESNESDT

IDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTV

QMRNSLSELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADAN

GAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

Table 2 shows ERCAS12A mutations with improved DNA cleavage activity in E. coli. The enrichment scores over 3 biological replicates were measured by deep sequencing. Mutants with average enrichment score greater than the maximal score of all synonymous variants (0.41) were considered as positive hits.

TABLE 2

ERCAS12A Mutations with Improved DNA

Cleavage Activity in E. coli

Enrich.

WT
Enrich.
Enrich.
Enrich.
Enrich.
(Std

No.
Mutant
Posit.
(REP1)
(REP 2)
(REP3)
(Avg)
Dev)

1
M1G
1
0.23
1.37
0.7
0.77
0.47

2
N2C
2
0.23
0.28
2.21
0.9
0.92

3
N3I
3
0.63
0.68
0.01
0.44
0.31

4
T5D
5
2.31
0.98
0.7
1.33
0.7

5
T5V
5
0.74
0.8
0.7
0.75
0.04

6
T5F
5
0.52
0.69
0.42
0.54
0.11

7
N6P
6
0.7
0.62
0.8
0.71
0.07

8
N7A
7
1.53
1.26
0.99
1.26
0.22

9
N7Q
7
1.01
1.14
0.83
1
0.13

10
N7E
7
0.64
0.4
1.07
0.7
0.28

11
N7I
7
0.87
0.73
0.45
0.68
0.17

12
N7D
7
0.64
0.69
0.7
0.68
0.03

13
N7Y
7
0.49
0.55
0.65
0.56
0.07

14
F8K
8
1.34
2.08
1.96
1.79
0.33

15
F8H
8
0.64
0.51
0.24
0.47
0.17

16
Q9D
9
0.24
1.39
0.71
0.78
0.47

17
N10A
10
1.09
0.98
0.7
0.92
0.16

18
NI0E
10
0.24
1.67
0.01
0.64
0.74

19
F11Q
11
0.24
0.98
0.86
0.69
0.33

20
F11E
11
0.93
0.29
0.41
0.54
0.28

21
I12Q
12
0.64
1.13
0.52
0.76
0.27

22
I12L
12
0.53
0.28
0.45
0.42
0.1

23
I14H
14
1.06
0.78
1.08
0.97
0.14

24
I14E
14
1.2
0.9
0.68
0.93
0.21

25
I14N
14
0.93
0.86
0.87
0.89
0.03

26
I14L
14
0.74
0.46
0.47
0.56
0.13

27
I14V
14
0.39
0.48
0.79
0.55
0.17

28
S15M
15
1.6
1.36
1.89
1.62
0.22

29
S15K
15
1.75
1.11
0.49
1.12
0.51

30
S15T
15
0.72
0.67
0.93
0.77
0.11

31
S16H
16
1.82
1.77
1.08
1.56
0.34

32
S16M
16
1.02
1.14
1.01
1.06
0.06

33
S16N
16
0.72
1.11
−0.42
0.47
0.65

34
Q18I
18
1.83
1.96
2.11
1.97
0.12

35
Q18V
18
1.05
1.62
1.83
1.5
0.33

36
Q18M
18
0.82
0.84
1.04
0.9
0.1

37
Q18T
18
1.22
0.73
0.38
0.77
0.34

38
Q18N
18
0.41
0.55
0.69
0.55
0.12

39
Q18W
18
1.05
0.95
−0.43
0.52
0.68

40
Q18R
18
0.42
0.55
0.3
0.42
0.11

41
A24E
24
0.64
0.62
0.35
0.54
0.13

42
I26P
26
1.53
1.14
1.26
1.31
0.16

43
P27F
27
2.41
2.51
2.51
2.48
0.05

44
P27M
27
1.93
1.89
1.94
1.92
0.02

45
P27Y
27
1.79
1.29
1.7
1.6
0.22

46
P27G
27
1.41
1.38
1.64
1.48
0.12

47
P27H
27
1.47
1.17
1.32
1.32
0.12

48
P27L
27
0.96
0.94
0.99
0.97
0.02

49
P27C
27
0.66
0.62
0.5
0.59
0.07

50
P27K
27
0.87
0.29
0.37
0.51
0.26

51
P27R
27
0.49
0.42
0.36
0.42
0.05

52
T28G
28
0.57
0.61
0.43
0.53
0.07

53
T30F
30
2.47
2.16
2.14
2.26
0.15

54
T30W
30
1.88
1.66
1.5
1.68
0.16

55
T30H
30
1.91
1.89
0.79
1.53
0.52

56
T30D
30
1.42
0.62
0.86
0.97
0.34

57
T30Q
30
0.62
0.35
0.7
0.56
0.15

58
T30V
30
0.29
0.53
0.42
0.42
0.1

59
T31G
31
2.57
2.65
2.71
2.65
0.06

60
T31C
31
2.38
2.73
2.62
2.58
0.15

61
T31N
31
2.13
2.06
2.3
2.16
0.1

62
T31A
31
1.9
2.01
1.92
1.94
0.05

63
T31P
31
1.04
1.03
1.02
1.03
0.01

64
T31M
31
0.5
0.63
0.71
0.62
0.09

65
T31L
31
0.35
0.5
0.66
0.51
0.13

66
T31H
31
0.63
0.58
0.31
0.5
0.14

67
T31S
31
0.44
0.58
0.41
0.48
0.07

68
T31D
31
0.53
0.15
0.71
0.46
0.23

69
Q32W
32
2.47
2.62
2.72
2.61
0.1

70
Q32M
32
1.89
2.07
2
1.99
0.07

71
Q32V
32
1.78
1.92
1.87
1.86
0.06

72
Q32F
32
1.78
1.81
1.92
1.84
0.06

73
Q32I
32
1.7
1.87
1.77
1.78
0.07

74
Q32C
32
1.65
1.68
1.83
1.72
0.08

75
Q32G
32
1.52
1.61
1.76
1.63
0.1

76
Q32T
32
1.38
1.31
1.43
1.38
0.05

77
Q32A
32
1.12
1.4
1.34
1.29
0.12

78
Q32L
32
1.21
1.32
1.29
1.27
0.05

79
Q32S
32
1.22
1.27
1.28
1.25
0.03

80
Q32Y
32
1.05
0.78
1.05
0.96
0.13

81
Q32N
32
0.91
0.72
0.79
0.81
0.08

82
Q32K
32
0.67
0.77
0.71
0.71
0.04

83
Q32P
32
0.51
0.52
0.52
0.52
0.01

84
Q33P
33
1.24
1.44
1.41
1.37
0.09

85
Q33F
33
1.12
1.13
1.34
1.2
0.1

86
Q33W
33
0.94
1.5
0.57
1.01
0.38

87
Q33V
33
0.86
0.89
1.04
0.93
0.08

88
Q33Y
33
0.42
0.89
0.94
0.75
0.23

89
Q33I
33
0.62
0.47
0.96
0.68
0.2

90
F34P
34
2.4
2.47
2.57
2.48
0.07

91
F34W
34
0.84
0.82
0.87
0.84
0.02

92
F34L
34
0.54
0.73
0.7
0.65
0.08

93
F34R
34
0.57
0.69
0.38
0.55
0.13

94
F34A
34
0.66
0.39
0.58
0.54
0.11

95
I35L
35
3.18
3.38
3.39
3.32
0.1

96
I35C
35
3.05
3.16
3.31
3.18
0.11

97
I35Q
35
2.63
2.63
2.9
2.72
0.13

98
I35A
35
2.48
2.56
2.69
2.58
0.09

99
I35S
35
2.48
2.49
2.54
2.5
0.03

100
I35E
35
2.56
2.37
2.51
2.48
0.08

101
I35M
35
1.91
2.14
2.1
2.05
0.1

102
I35N
35
1.81
1.79
1.84
1.81
0.02

103
I35F
35
1.68
1.54
1.84
1.69
0.12

104
I35T
35
1.36
1.4
1.46
1.41
0.04

105
I35Y
35
1.26
1.38
1.52
1.38
0.11

106
I35W
35
1.06
1.27
1
1.11
0.12

107
I35K
35
1.13
0.97
1.22
1.11
0.1

108
I35V
35
0.59
0.7
0.9
0.73
0.13

109
I35P
35
0.38
0.51
0.66
0.52
0.11

110
I35G
35
0.55
0.16
0.72
0.48
0.23

111
V36P
36
1.7
1.88
1.96
1.85
0.11

112
V36K
36
1.65
1.58
1.84
1.69
0.11

113
V36M
36
0.47
0.67
0.59
0.58
0.08

114
K37P
37
1.86
2.01
1.99
1.95
0.06

115
N38L
38
1.96
2.04
2
2
0.03

116
N38R
38
1.58
1.8
1.75
1.71
0.1

117
N38G
38
1.52
1.73
1.83
1.69
0.13

118
N38D
38
1.5
1.64
1.93
1.69
0.18

119
N38P
38
1.37
1.54
1.65
1.52
0.11

120
N38A
38
1.27
1.39
1.38
1.35
0.06

121
N38T
38
1.19
1.18
1.03
1.13
0.07

122
N38H
38
0.84
1.06
0.95
0.95
0.09

123
N38M
38
0.9
0.8
0.82
0.84
0.04

124
N38S
38
0.77
0.92
0.76
0.81
0.08

125
N38Q
38
0.72
0.8
0.42
0.65
0.16

126
N38I
38
0.51
0.64
0.61
0.59
0.05

127
N38E
38
0.62
0.36
0.48
0.49
0.1

128
N38V
38
0.47
0.45
0.52
0.48
0.03

129
G39Q
39
3.42
3.64
3.58
3.55
0.09

130
G39K
39
3.24
3.42
3.48
3.38
0.1

131
G39M
39
2.99
3.15
3.13
3.09
0.07

132
G39A
39
2.97
3.12
3.15
3.08
0.08

133
G39V
39
2.9
3.13
3.18
3.07
0.12

134
G39H
39
2.87
2.99
2.88
2.91
0.05

135
G39E
39
2.75
2.95
2.96
2.89
0.1

136
G39F
39
2.56
2.93
2.87
2.79
0.16

137
G39N
39
2.65
2.81
2.85
2.77
0.09

138
G39I
39
2.66
2.79
2.82
2.76
0.07

139
G39L
39
2.59
2.76
2.81
2.72
0.09

140
G39T
39
2.41
2.53
2.53
2.49
0.06

141
G39W
39
2.2
2.41
2.31
2.31
0.09

142
G39C
39
2.12
2.12
2.14
2.13
0.01

143
G39S
39
2.05
2.06
2.19
2.1
0.06

144
G39Y
39
1.83
1.98
1.85
1.89
0.06

145
G39P
39
0.49
1.05
1.13
0.89
0.28

146
G39D
39
0.52
0.74
0.92
0.73
0.17

147
G39R
39
0.61
0.75
0.73
0.7
0.06

148
I40Q
40
2.91
3
3.08
3
0.07

149
I40N
40
2.6
2.81
2.77
2.73
0.09

150
I40Y
40
2.49
2.62
2.78
2.63
0.12

151
I40R
40
2.55
2.62
2.71
2.62
0.06

152
I40E
40
2.38
2.55
2.66
2.53
0.12

153
I40C
40
2.38
2.51
2.48
2.46
0.06

154
I40V
40
2.29
2.52
2.5
2.44
0.11

155
I40T
40
2.39
2.45
2.44
2.42
0.03

156
I40K
40
1.59
1.76
1.75
1.7
0.08

157
I40D
40
1.42
1.61
1.6
1.55
0.09

158
I40W
40
1.38
1.43
1.63
1.48
0.11

159
I40L
40
1.13
1.25
1.24
1.2
0.05

160
I40P
40
0.62
0.83
0.81
0.75
0.1

161
E43T
43
1.14
1.32
1.16
1.21
0.08

162
E43Q
43
0.92
0.98
1.19
1.03
0.11

163
E43S
43
0.73
0.98
0.81
0.84
0.11

164
E43L
43
0.85
0.71
0.71
0.75
0.07

165
E43I
43
0.77
0.76
0.69
0.74
0.04

166
E43V
43
0.65
0.68
0.62
0.65
0.02

167
E43C
43
0.38
0.73
0.65
0.59
0.15

168
E45R
45
0.96
1.13
0.66
0.92
0.19

169
E45F
45
0.52
0.57
0.74
0.61
0.09

170
L46N
46
1.25
1.4
1.6
1.42
0.15

171
L46S
46
1.29
1.55
1.41
1.41
0.11

172
L46D
46
1.12
1.35
1.31
1.26
0.1

173
L46K
46
0.93
0.87
1.23
1.01
0.16

174
L46G
46
0.79
0.76
1.05
0.87
0.13

175
L46C
46
0.79
0.85
0.64
0.76
0.09

176
L46H
46
0.75
0.48
0.91
0.71
0.17

177
L46T
46
0.6
0.7
0.79
0.7
0.08

178
L46E
46
0.57
0.56
0.55
0.56
0.01

179
E49Y
49
1.83
1.88
1.77
1.82
0.04

180
E49N
49
1.47
1.52
1.86
1.61
0.18

181
E49P
49
1.57
1.64
1.33
1.51
0.14

182
E49F
49
1.21
1.18
1.31
1.23
0.05

183
E49R
49
1.17
1.26
1.26
1.23
0.04

184
E49M
49
1.29
1.06
0.82
1.06
0.19

185
E49T
49
0.83
1.34
0.86
1.01
0.23

186
E49H
49
1.01
1.16
0.63
0.93
0.22

187
E49L
49
0.86
0.82
0.85
0.84
0.02

188
E49C
49
0.46
0.91
0.83
0.73
0.2

189
E49I
49
0.92
0.44
0.69
0.69
0.2

190
E49S
49
0.63
0.53
0.65
0.6
0.05

191
E49K
49
0.43
0.4
0.42
0.42
0.01

192
N50Q
50
0.51
0.11
0.91
0.51
0.33

193
Q52V
52
0.61
0.65
0.69
0.65
0.03

194
Q52C
52
0.62
0.58
0.42
0.54
0.09

195
L54G
54
0.59
0.55
0.51
0.55
0.03

196
D56P
56
1.15
1.41
1.52
1.36
0.15

197
D56R
56
0.98
0.95
0.95
0.96
0.01

198
D56K
56
0.61
1.03
0.58
0.74
0.21

199
D56A
56
0.57
0.29
0.68
0.51
0.16

200
I57E
57
1.3
1.28
1.3
1.3
0.01

201
I57C
57
1.48
1.27
0.89
1.21
0.25

202
I57P
57
0.95
0.93
0.84
0.91
0.05

203
I57G
57
0.74
0.65
0.98
0.79
0.14

204
I57A
57
0.91
0.59
0.86
0.79
0.14

205
I57T
57
0.47
0.66
0.55
0.56
0.08

206
I57Q
57
0.94
0.36
0.37
0.56
0.27

207
I57V
57
0.53
0.58
0.38
0.5
0.09

208
I57L
57
0.32
0.56
0.37
0.42
0.1

209
D59A
59
0.94
1.09
1.11
1.05
0.08

210
D59I
59
0.95
1.07
0.65
0.89
0.17

211
D59S
59
0.76
0.92
0.93
0.87
0.08

212
D59T
59
0.78
0.98
0.66
0.81
0.13

213
D59R
59
0.93
0.65
0.62
0.73
0.14

214
D59G
59
0.86
0.54
0.62
0.67
0.14

215
D59M
59
0.49
0.7
0.4
0.53
0.13

216
D59L
59
0.42
0.52
0.56
0.5
0.06

217
D59Q
59
0.67
0.29
0.38
0.45
0.16

218
D60R
60
0.92
1.26
1.38
1.18
0.19

219
D60N
60
1.23
0.68
1.22
1.04
0.26

220
D60M
60
0.76
0.82
0.77
0.78
0.02

221
D60T
60
0.48
0.89
0.83
0.73
0.18

222
D60G
60
0.6
0.73
0.85
0.73
0.1

223
D60V
60
0.92
0.74
0.39
0.69
0.22

224
D60K
60
0.9
0.4
0.61
0.64
0.2

225
D60Y
60
0.34
0.72
0.73
0.6
0.18

226
D60P
60
0.65
0.76
0.36
0.59
0.17

227
D60F
60
0.87
−0.02
0.75
0.53
0.4

228
D60L
60
0.48
0.75
0.25
0.49
0.2

229
D60A
60
0.51
0.32
0.62
0.49
0.12

230
Y62Q
62
1.41
1.96
1.06
1.48
0.37

231
Y62A
62
1.37
0.78
0.85
1
0.26

232
Y62F
62
1.2
0.67
0.98
0.95
0.22

233
Y62V
62
0.63
1.04
0.79
0.82
0.17

234
Y62N
62
0.29
0.79
0.28
0.45
0.24

235
R63M
63
0.49
0.69
0.41
0.53
0.12

236
R63V
63
0.4
0.56
0.34
0.43
0.09

237
F65E
65
1.27
1.08
0.8
1.05
0.19

238
F65K
65
0.79
0.4
0.75
0.65
0.17

239
F65R
65
0.45
0.47
0.44
0.46
0.01

240
S67V
67
1.04
0.68
0.92
0.88
0.15

241
S67Y
67
0.77
0.77
0.68
0.74
0.04

242
S67F
67
0.76
0.84
0.56
0.72
0.12

243
S67Q
67
0.59
0.45
0.57
0.53
0.06

244
S67A
67
0.55
0.4
0.51
0.49
0.06

245
S67W
67
0.49
0.48
0.31
0.43
0.08

246
E68V
68
0.99
1.13
1.51
1.21
0.22

247
E68A
68
1.18
0.41
0.38
0.66
0.37

248
E68R
68
0.75
0.64
0.36
0.58
0.16

249
T69F
69
1.81
2.06
1.93
1.93
0.1

250
T69M
69
0.49
1.24
0.68
0.8
0.32

251
T69D
69
1.3
0.26
0.68
0.75
0.43

252
T69H
69
0.2
0.96
0.68
0.61
0.31

253
T69W
69
0.2
0.77
0.5
0.49
0.23

254
T69L
69
0.44
0.39
0.5
0.44
0.04

255
S71Q
71
0.61
1.28
0.39
0.76
0.38

256
I73E
73
0.38
0.5
0.56
0.48
0.07

257
D75R
75
0.81
0.27
0.84
0.64
0.26

258
I76Q
76
0.61
0.83
0.86
0.77
0.11

259
I76G
76
0.79
0.74
0.59
0.71
0.09

260
I76F
76
1.57
−0.12
0.5
0.65
0.7

261
D77P
77
0.75
0.64
0.66
0.68
0.05

262
T79E
79
0.52
0.39
0.56
0.49
0.07

263
T79R
79
0.45
0.53
0.39
0.46
0.05

264
S80N
80
0.62
0.5
0.31
0.48
0.13

265
S80Q
80
0.44
0.62
0.32
0.46
0.12

266
S80V
80
0.36
0.42
0.48
0.42
0.05

267
L81Y
81
0.71
0.73
0.6
0.68
0.05

268
L81H
81
0.64
0.68
0.51
0.61
0.07

269
L81W
81
0.39
0.59
0.73
0.57
0.14

270
L81E
81
0.94
0.28
0.39
0.54
0.29

271
F82Q
82
0.38
0.42
0.66
0.49
0.12

272
K84F
84
0.4
0.66
0.49
0.52
0.11

273
M85G
85
1.06
1.18
1.18
1.14
0.06

274
M85Q
85
0.79
0.8
1.34
0.98
0.26

275
M85Y
85
1.11
0.78
0.94
0.95
0.14

276
M85A
85
0.85
0.79
1.09
0.91
0.13

277
M85N
85
0.67
0.98
0.82
0.82
0.13

278
M85W
85
0.56
0.44
1.11
0.7
0.29

279
M85E
85
0.7
0.51
0.64
0.62
0.08

280
M85F
85
0.42
0.58
0.42
0.47
0.08

281
E86N
86
0.44
1.02
0.66
0.71
0.24

282
I87G
87
0.7
0.79
0.46
0.65
0.14

283
I87A
87
0.68
0.55
0.29
0.51
0.16

284
I87H
87
0.25
0.65
0.62
0.5
0.18

285
I87W
87
0.38
0.46
0.43
0.42
0.03

286
Q88N
88
1.07
0.64
0.84
0.85
0.18

287
Q88E
88
0.28
1.1
0.97
0.78
0.36

288
Q88S
88
0.41
0.47
0.5
0.46
0.04

289
L89Q
89
0.97
0.96
1.21
1.05
0.12

290
L89S
89
0.47
0.54
0.6
0.54
0.05

291
L89C
89
0.6
0.61
0.37
0.52
0.11

292
L89D
89
0.63
0.32
0.47
0.48
0.13

293
L89G
89
0.21
0.8
0.33
0.45
0.25

294
K90P
90
1.13
1.38
1.11
1.21
0.12

295
N91V
91
0.84
0.37
0.47
0.56
0.2

296
G92D
92
0.21
1.04
0.37
0.54
0.36

297
G92F
92
−0.07
0
1.51
0.48
0.73

298
D93L
93
0.83
0.76
1.16
0.92
0.17

299
D93P
93
0.68
0.87
0.7
0.75
0.09

300
D93T
93
0.46
0.45
0.96
0.62
0.24

301
D93N
93
−0.04
0.65
0.81
0.47
0.37

302
D93R
93
0.36
0.36
0.66
0.46
0.14

303
D93M
93
0.68
0.66
0.01
0.45
0.31

304
N94P
94
1.28
0.92
0.59
0.93
0.28

305
K95T
95
0.8
1
1.03
0.94
0.1

306
K95F
95
0.5
0.8
0.95
0.75
0.19

307
K95L
95
0.9
0.62
0.48
0.67
0.17

308
K95P
95
0.82
0.69
0.44
0.65
0.16

309
K95E
95
0.97
0.23
0.64
0.61
0.3

310
K95W
95
0.43
0.46
0.61
0.5
0.08

311
K95I
95
0.64
0.41
0.4
0.48
0.11

312
K95S
95
0.4
0.55
0.37
0.44
0.08

313
D96C
96
0.91
0.51
0.57
0.66
0.18

314
T97W
97
0.95
0.29
0.42
0.55
0.28

315
L98Q
98
1.8
1.34
1.71
1.62
0.2

316
L98A
98
1.07
1.3
0.84
1.07
0.19

317
L98H
98
0.92
0.92
1.12
0.99
0.09

318
L98D
98
1.27
0.83
0.85
0.98
0.2

319
L98N
98
0.92
0.99
0.74
0.88
0.1

320
L98G
98
0.76
0.81
0.82
0.8
0.03

321
L98S
98
0.82
0.84
0.62
0.76
0.1

322
L98T
98
0.88
0.77
0.41
0.69
0.2

323
L98C
98
0.84
0.3
0.76
0.63
0.24

324
L98P
98
0.53
0.41
0.36
0.43
0.07

325
I99W
99
0.96
1.23
0.8
1
0.18

326
I99Y
99
0.83
0.79
0.69
0.77
0.06

327
E101P
101
0.39
0.6
0.6
0.53
0.1

328
Q102I
102
0.92
0.3
2.73
1.31
1.03

329
Q102V
102
0.63
0.9
1
0.84
0.16

330
T103Y
103
0.63
0.7
0.42
0.58
0.12

331
E104P
104
0.6
0.82
0.86
0.76
0.11

332
Y105R
105
1.49
1.87
1.86
1.74
0.18

333
Y105P
105
1.04
0.52
1.89
1.15
0.57

334
Y105A
105
0.63
0.29
1.52
0.82
0.52

335
Y105T
105
1
0.11
0.87
0.66
0.39

336
Y105G
105
0.22
0.52
0.94
0.56
0.29

337
R106T
106
0.79
0.3
0.71
0.6
0.21

338
R106V
106
0.29
0.65
0.52
0.49
0.15

339
R106I
106
0.23
0.7
0.38
0.44
0.2

340
A108R
108
0.73
0.43
0.41
0.52
0.15

341
K111T
111
0.75
0.63
0.78
0.72
0.07

342
K111W
111
0.62
0.71
0.59
0.64
0.05

343
K111G
111
0.6
0.61
0.69
0.63
0.04

344
K111A
111
0.56
0.69
0.56
0.6
0.06

345
K111F
111
0.44
0.59
0.7
0.57
0.11

346
K111V
111
0.69
0.51
0.43
0.54
0.11

347
K111S
111
0.48
0.62
0.52
0.54
0.06

348
K111C
111
0.32
0.39
0.56
0.42
0.1

349
K112D
112
1.73
0.3
2.17
1.4
0.8

350
K112H
112
0.92
1.76
0.72
1.14
0.45

351
K112E
112
0.99
1.12
1.03
1.05
0.06

352
K112L
112
0.79
1.4
0.43
0.87
0.4

353
K112T
112
0.52
0.81
0.87
0.73
0.16

354
K112C
112
0.63
0.3
0.72
0.55
0.18

355
K112R
112
0.55
0.13
0.96
0.55
0.34

356
K112P
112
0.41
0.63
0.5
0.51
0.09

357
F113A
113
0.23
0.42
0.72
0.45
0.2

358
A114S
114
1.21
1.28
0.72
1.07
0.25

359
A114E
114
1.07
−0.11
0.87
0.61
0.52

360
N115W
115
0.7
0.99
0.02
0.57
0.4

361
N115M
115
0.42
0.25
0.78
0.48
0.22

362
R118D
118
2.53
2.24
1.81
2.19
0.29

363
R118A
118
0.92
0.99
0.93
0.95
0.03

364
R118Y
118
1.11
−0.04
0.47
0.51
0.47

365
R118N
118
0.92
1.21
−0.68
0.48
0.83

366
R118V
118
0.85
−0.17
0.65
0.44
0.44

367
F119H
119
1.48
1.21
1.4
1.36
0.11

368
F119D
119
0.45
0.29
0.71
0.48
0.17

369
K120A
120
0.95
0.88
0.57
0.8
0.17

370
K120C
120
0.36
0.29
0.67
0.44
0.16

371
K120V
120
0.19
0.45
0.66
0.43
0.19

372
K120T
120
0.34
0.32
0.63
0.43
0.14

373
K120P
120
0.28
0.55
0.44
0.42
0.11

374
N121F
121
0.48
0.32
0.53
0.45
0.09

375
M122W
122
0.51
0.47
0.45
0.48
0.02

376
F123E
123
0.44
0.43
0.48
0.45
0.02

377
S124N
124
1.32
0.38
0.11
0.6
0.52

378
L127Q
127
0.4
0.46
1.34
0.73
0.43

379
I128Q
128
2.16
1.89
1.39
1.81
0.32

380
S129G
129
0.91
1.38
1.1
1.13
0.19

381
S129F
129
0.21
1.38
1.1
0.9
0.5

382
S129D
129
−0.48
0.97
0.92
0.47
0.67

383
S129I
129
0.91
0.41
0.01
0.44
0.37

384
D130I
130
0.47
0.67
0.22
0.45
0.18

385
L132T
132
0.91
1.2
1.11
1.07
0.12

386
L132G
132
0.73
0.95
0.75
0.81
0.1

387
L132A
132
0.64
0.8
0.76
0.73
0.07

388
F135D
135
1.04
0.82
0.63
0.83
0.17

389
F135A
135
0.6
0.67
0.53
0.6
0.06

390
F135H
135
0.55
0.46
0.54
0.52
0.04

391
F135T
135
0.35
0.6
0.33
0.43
0.12

392
V136Y
136
0.59
0.65
0.32
0.52
0.15

393
I137Q
137
0.49
0.73
0.49
0.57
0.11

394
H138M
138
0.9
1.58
0.52
1
0.44

395
H138A
138
0.21
1.07
0.88
0.72
0.37

396
H138S
138
0.21
0.81
0.91
0.64
0.31

397
H138V
138
0.97
0.53
0.14
0.55
0.34

398
N139Q
139
1.72
2.15
2.61
2.16
0.36

399
N139C
139
1.42
1.13
1.39
1.31
0.13

400
N139A
139
0.72
0.68
1.43
0.95
0.34

401
N139R
139
0.93
1.05
0.36
0.78
0.3

402
N139I
139
0.79
0.28
0.9
0.65
0.27

403
N139P
139
0.21
−0.13
1.68
0.59
0.78

404
N139L
139
0.29
0.53
0.52
0.45
0.11

405
N139F
139
0.72
0.28
0.29
0.43
0.21

406
N139G
139
0.76
0.18
0.31
0.42
0.25

407
N140C
140
1.92
1.89
2.02
1.94
0.06

408
N140G
140
0.71
0.9
0.66
0.76
0.1

409
N140P
140
0.81
0.83
0.6
0.75
0.1

410
N140T
140
0.68
0.84
0.51
0.68
0.14

411
N140S
140
0.58
0.54
0.62
0.58
0.03

412
N141P
141
0.51
0.06
1.04
0.54
0.4

413
N141F
141
0.1
0.53
0.75
0.46
0.27

414
Y142G
142
0.77
0.69
0.85
0.77
0.06

415
Y142K
142
0.77
0.89
0.52
0.73
0.15

416
Y142E
142
0.27
0.44
1.23
0.65
0.42

417
Y142R
142
0.74
0.86
0.33
0.64
0.22

418
Y142P
142
0.82
0.28
0.66
0.59
0.22

419
Y142V
142
0.54
0.61
0.19
0.45
0.19

420
S143R
143
0.36
0.47
1.68
0.84
0.6

421
S143E
143
0.7
0.69
0.42
0.6
0.13

422
S143G
143
0.91
0.38
0.35
0.55
0.26

423
S143V
143
0.77
0.65
0.17
0.53
0.26

424
S143I
143
0.41
0.83
0.12
0.45
0.29

425
A144R
144
0.83
1.42
2.34
1.53
0.62

426
A144I
144
1.31
0.29
1.96
1.19
0.69

427
A144K
144
1.21
0.54
1.06
0.94
0.29

428
A144N
144
0.21
0.8
0.86
0.62
0.29

429
A144P
144
0.37
0.29
0.93
0.53
0.28

430
A144C
144
0.42
0.58
0.59
0.53
0.08

431
A144D
144
−0.7
0.98
1.23
0.5
0.86

432
A144T
144
0.83
0.21
0.2
0.42
0.29

433
E146R
146
1.36
0.54
3.04
1.65
1.04

434
E146T
146
1.75
1.38
1.11
1.42
0.26

435
E146M
146
0.77
1.1
0.42
0.76
0.28

436
E146I
146
0.62
0.51
1.11
0.75
0.26

437
E146S
146
0.55
0.47
0.67
0.56
0.08

438
K147F
147
0.68
0.82
1.11
0.87
0.18

439
K147A
147
0.4
1.24
0.48
0.71
0.38

440
K147G
147
0.68
0.62
0.8
0.7
0.07

441
K147Q
147
0.09
0.11
1.86
0.69
0.83

442
K147L
147
0.74
0.74
0.52
0.66
0.1

443
E148C
148
0.43
0.45
1.54
0.81
0.52

444
E148P
148
0.82
0.1
0.42
0.45
0.29

445
E149L
149
0.99
0.98
0.62
0.86
0.17

446
E149R
149
0.59
0.93
0.18
0.57
0.31

447
K150G
150
1.87
1.2
1.67
1.58
0.28

448
K150C
150
1.31
1.38
1.11
1.27
0.12

449
K150A
150
1.39
0.84
0.82
1.02
0.26

450
K150I
150
−0.48
1.49
1.36
0.79
0.9

451
K150W
150
1.06
0.03
0.81
0.64
0.44

452
K150V
150
0.22
1.18
0.32
0.57
0.43

453
K150L
150
0.5
0.69
0.45
0.55
0.1

454
K150Y
150
0.29
−0.08
1.34
0.52
0.6

455
T151E
151
0.67
0.75
0.75
0.72
0.03

456
T151C
151
0.5
0.9
0.36
0.59
0.23

457
T151V
151
0.55
0.67
0.45
0.56
0.09

458
T151Q
151
0.47
0.42
0.63
0.51
0.09

459
V153K
153
0.49
0.85
0.84
0.73
0.17

460
I154P
154
1.04
1.04
0.96
1.01
0.04

461
I154E
154
1.07
0.5
0.74
0.77
0.23

462
I154A
154
0.72
0.41
0.55
0.56
0.13

463
I154Q
154
0.75
0.33
0.27
0.45
0.22

464
I154H
154
0.33
0.4
0.55
0.43
0.09

465
L156C
156
0.59
0.25
0.43
0.43
0.14

466
S158N
158
1.14
0.84
0.92
0.97
0.13

467
S158G
158
0.75
0.58
0.83
0.72
0.1

468
S158K
158
0.46
0.55
0.49
0.5
0.04

469
S158D
158
0.37
0.48
0.55
0.47
0.08

470
F160H
160
0.91
0.29
0.71
0.64
0.26

471
S163I
163
0.71
0.83
0.72
0.75
0.06

472
F164Q
164
0.55
1.08
0.36
0.67
0.31

473
K165Y
165
0.32
0.72
0.5
0.52
0.16

474
Y167M
167
0.66
0.86
0.64
0.72
0.1

475
Y167I
167
0.62
0.51
0.91
0.68
0.17

476
Y167L
167
0.46
0.47
0.54
0.49
0.03

477
K169C
169
0.81
0.78
0.87
0.82
0.03

478
K169F
169
0.7
0.64
0.65
0.66
0.03

479
A172Y
172
0.22
1
1.42
0.88
0.5

480
C174V
174
0.47
0.7
0.8
0.66
0.14

481
C174T
174
0.78
0.44
0.67
0.63
0.14

482
C174I
174
0.37
0.46
0.51
0.45
0.06

483
A177W
177
1
0.31
0.63
0.65
0.28

484
A177M
177
0.42
0.42
0.44
0.43
0.01

485
D178R
178
0.73
0.57
0.59
0.63
0.07

486
D179W
179
0.71
0.78
0.81
0.76
0.04

487
D179P
179
0.34
0.23
0.7
0.42
0.2

488
S181N
181
0.38
0.39
0.99
0.58
0.29

489
S181H
181
0.84
0.76
−0.04
0.52
0.4

490
S181V
181
0.42
0.38
0.54
0.45
0.07

491
S184I
184
1.57
1.58
1.65
1.6
0.03

492
S184M
184
1.34
1
1.36
1.23
0.16

493
S184V
184
0.59
0.64
0.75
0.66
0.07

494
S184L
184
0.34
0.52
0.5
0.45
0.08

495
S184C
184
0.57
0.52
0.2
0.43
0.16

496
I188W
188
1.14
0.72
0.72
0.86
0.2

497
I188A
188
0.86
0.59
0.88
0.78
0.13

498
I188C
188
0.6
0.52
0.59
0.57
0.04

499
N190Q
190
1.14
0.31
0.91
0.79
0.35

500
N190R
190
0.36
1.51
0.25
0.71
0.57

501
N192G
192
0.69
0.71
0.79
0.73
0.05

502
N192C
192
0.84
0.16
0.35
0.45
0.29

503
A193H
193
0.8
0.74
0.92
0.82
0.07

504
A193F
193
0.73
0.64
0.58
0.65
0.06

505
A193L
193
0.56
0.6
0.7
0.62
0.06

506
A193N
193
0.36
0.66
0.62
0.55
0.13

507
A193Q
193
0.42
0.65
0.55
0.54
0.09

508
I195W
195
0.64
0.57
0.46
0.56
0.07

509
F196T
196
0.57
0.27
0.52
0.45
0.13

510
F197E
197
0.63
0.73
0.66
0.67
0.05

511
F197R
197
0.61
0.58
0.5
0.56
0.05

512
F197K
197
0.58
0.48
0.62
0.56
0.06

513
F197H
197
0.38
0.59
0.42
0.46
0.09

514
F197Q
197
0.45
0.24
0.6
0.43
0.15

515
S198D
198
0.78
0.59
0.64
0.67
0.08

516
S198E
198
0.28
0.68
0.56
0.51
0.17

517
S198K
198
0.14
0.72
0.66
0.51
0.26

518
S198N
198
−0.25
0.49
1.27
0.5
0.62

519
A200H
200
1.32
1.42
1.44
1.39
0.05

520
A200I
200
0.22
1.11
0.52
0.62
0.37

521
V202H
202
1.83
2.1
0.73
1.55
0.59

522
V202C
202
0.44
0.63
0.67
0.58
0.1

523
R205M
205
0.72
1.61
1.65
1.33
0.43

524
R205T
205
0.37
0.6
0.44
0.47
0.1

525
R205C
205
0.69
0.26
0.32
0.42
0.19

526
I206Y
206
0.72
0.75
1.08
0.85
0.16

527
I206W
206
1.1
0.1
0.35
0.52
0.43

528
I206D
206
0.99
0.13
0.44
0.52
0.36

529
V207W
207
0.42
0.4
0.54
0.46
0.06

530
S209K
209
0.57
0.51
0.63
0.57
0.05

531
L210N
210
0.57
0.23
0.57
0.46
0.16

532
D214T
214
0.36
0.62
0.76
0.58
0.17

533
D214W
214
0.49
0.5
0.56
0.51
0.03

534
I215W
215
1.13
0.78
0.37
0.76
0.31

535
I215A
215
0.26
0.31
0.73
0.43
0.21

536
N216F
216
0.62
0.25
0.67
0.51
0.19

537
K217A
217
0.4
0.49
1.2
0.69
0.36

538
K217D
217
0.4
0.64
0.73
0.59
0.14

539
K217V
217
0.52
0.55
0.58
0.55
0.02

540
S219P
219
0.21
1.16
0.88
0.75
0.4

541
S219F
219
−0.07
0.71
0.85
0.5
0.41

542
S219W
219
1.24
−0.2
0.37
0.47
0.59

543
S219D
219
0.12
0.62
0.63
0.46
0.24

544
G220Y
220
0.21
1
0.73
0.65
0.33

545
D221N
221
0.91
1
1.42
1.11
0.22

546
D221I
221
0.21
1
1.64
0.95
0.58

547
D221C
221
1.31
1.26
0.03
0.87
0.59

548
D221R
221
0.57
0.31
0.73
0.53
0.17

549
D221P
221
1
0.08
0.5
0.53
0.38

550
D221L
221
0.5
0.51
0.4
0.47
0.05

551
D221S
221
−0.14
0.78
0.62
0.42
0.4

552
M222P
222
0.56
0.54
0.49
0.53
0.03

553
M222C
222
1.02
0.71
−0.44
0.43
0.63

554
K223F
223
1.92
1.81
1.13
1.62
0.35

555
K223P
223
1.42
1
1.5
1.3
0.22

556
K223C
223
0.91
1.91
0.73
1.18
0.52

557
K223G
223
0.97
0.15
0.65
0.59
0.34

558
D224H
224
−0.48
0.71
1.13
0.45
0.68

559
S225A
225
0.91
0.31
0.44
0.55
0.26

560
S225Y
225
0.91
1.41
−0.66
0.55
0.88

561
S225W
225
0.8
0.31
0.5
0.54
0.2

562
L226D
226
1.06
1.51
0.73
1.1
0.32

563
L226Y
226
0.83
0.84
1.22
0.97
0.18

564
E228P
228
0.92
0.37
0.71
0.67
0.23

565
E228F
228
0.45
0.47
0.61
0.51
0.07

566
E228W
228
0.25
0.44
0.71
0.47
0.19

567
E228T
228
0.4
0.49
0.43
0.44
0.04

568
M229S
229
0.46
0.64
0.34
0.48
0.12

569
S230G
230
0.87
0.38
0.6
0.61
0.2

570
S230E
230
0.84
0.3
0.66
0.6
0.22

571
L231K
231
0.85
0.89
0.62
0.79
0.12

572
L231D
231
0.55
0.64
1.13
0.77
0.26

573
L231R
231
1.04
0.4
0.86
0.77
0.27

574
E233S
233
0.76
−0.21
0.98
0.51
0.52

575
I234W
234
0.9
0.7
1.28
0.96
0.24

576
I234G
234
1.04
0.65
−0.13
0.52
0.48

577
Y235N
235
0.21
1.22
1.21
0.88
0.47

578
Y235L
235
1.49
0.7
0.29
0.83
0.5

579
Y235W
235
1.17
0.3
0.62
0.69
0.36

580
Y235A
235
0.9
−0.11
1.13
0.64
0.54

581
Y235T
235
0.72
−0.11
0.88
0.5
0.43

582
Y235Q
235
0.21
1.22
0.03
0.48
0.52

583
S236K
236
1.09
0.48
1.19
0.92
0.31

584
S236V
236
0.9
0.99
0.72
0.87
0.11

585
S236W
236
0.56
0.86
0.59
0.67
0.14

586
S236N
236
1.24
0.99
−0.48
0.58
0.76

587
S236G
236
0.55
0.52
0.29
0.45
0.12

588
E238F
238
0.37
0.51
1.15
0.68
0.34

589
E238S
238
0.46
0.75
0.57
0.59
0.12

590
K239T
239
1.27
0.7
0.47
0.81
0.33

591
K239V
239
0.62
0.39
0.61
0.54
0.1

592
K239W
239
0.57
0.55
0.48
0.53
0.04

593
K239F
239
0.33
0.16
0.89
0.46
0.31

594
K239G
239
0.5
0.12
0.72
0.44
0.25

595
G241P
241
2.15
0.66
0.38
1.06
0.77

596
G241M
241
1.07
1.2
0.76
1.01
0.18

597
G241Q
241
0.84
1.17
0.78
0.93
0.17

598
G241W
241
0.18
0.55
1.05
0.6
0.36

599
G241H
241
0.39
0.4
0.77
0.52
0.18

600
G241F
241
0.6
0.72
0.15
0.49
0.25

601
F243R
243
1.02
0.99
1.04
1.02
0.02

602
F243H
243
0.21
1.28
0.18
0.55
0.51

603
I244Y
244
1.22
1.31
0.84
1.12
0.2

604
I244Q
244
1.39
0.01
1.2
0.87
0.61

605
I244G
244
0.8
0.83
0.61
0.75
0.09

606
I244F
244
0.69
0.53
0.45
0.55
0.1

607
I244W
244
0.72
0.4
0.31
0.48
0.18

608
T245Q
245
1.41
1.14
1.41
1.32
0.13

609
T245E
245
0.9
0.99
1.12
1
0.09

610
T245L
245
1.15
0.61
1.2
0.99
0.27

611
T245Y
245
1.17
0.16
0.83
0.72
0.42

612
T245V
245
0.31
0.74
0.46
0.5
0.17

613
T245K
245
0.03
0.58
0.8
0.47
0.32

614
E247T
247
1.12
0.37
1.2
0.9
0.37

615
E247M
247
0.88
0.57
0.51
0.65
0.16

616
E247C
247
0.9
0.85
−0.11
0.54
0.47

617
E247I
247
0.7
0.67
−0.02
0.45
0.33

618
G248M
248
0.86
0.48
0.71
0.68
0.16

619
G248K
248
0.35
0.51
0.51
0.46
0.07

620
S250N
250
0.61
0.96
0.94
0.84
0.16

621
N253Q
253
1.71
1.21
1.41
1.44
0.21

622
N253G
253
1.12
1.54
0.72
1.13
0.34

623
N253W
253
1.12
0.7
0.58
0.8
0.23

624
N253A
253
1.06
0.29
1
0.78
0.35

625
D254Q
254
1.31
0.98
1.41
1.23
0.18

626
D254S
254
1.01
0.94
0.96
0.97
0.03

627
D254W
254
1.31
0.29
1.04
0.88
0.43

628
D254I
254
1.46
0.29
0.03
0.59
0.62

629
D254R
254
0.03
0.7
0.87
0.53
0.36

630
D254M
254
1.53
0.7
−0.67
0.52
0.91

631
C256Q
256
0.43
0.74
0.62
0.6
0.13

632
C256H
256
0.56
0.3
0.77
0.54
0.19

633
C256E
256
0.4
0.61
0.46
0.49
0.09

634
C256N
256
0.6
0.3
0.51
0.47
0.13

635
C256M
256
0.49
0.54
0.38
0.47
0.07

636
K258L
258
0.69
0.52
0.3
0.51
0.16

637
V259Y
259
1.81
1.39
1.41
1.54
0.2

638
V259Q
259
0.21
0.29
1.63
0.71
0.65

639
V259N
259
0.9
0.29
0.72
0.63
0.26

640
S261H
261
0.9
0.29
0.87
0.68
0.28

641
S261F
261
0.36
1.06
0.18
0.53
0.38

642
F262R
262
1.67
1.83
1.22
1.57
0.26

643
F262P
262
1.59
0.28
1.12
1
0.54

644
F262K
262
1.45
0.51
0.58
0.85
0.43

645
F262G
262
0.42
0.69
1.03
0.71
0.25

646
M263E
263
0.87
0.41
0.64
0.64
0.19

647
M263C
263
0.64
0.63
0.53
0.6
0.05

648
M263S
263
0.53
0.45
0.27
0.42
0.11

649
L265W
265
1.59
−0.3
0.89
0.73
0.78

650
L265K
265
0.61
0.59
0.43
0.54
0.08

651
Y266R
266
0.2
3.72
0.71
1.54
1.55

652
Y266E
266
0.81
0.79
0.31
0.64
0.23

653
Y266P
266
0.9
−0.12
0.53
0.43
0.42

654
C267K
267
1.12
1.2
1.63
1.31
0.22

655
C267M
267
0.61
1.38
0.65
0.88
0.36

656
C267E
267
0.89
1.38
0.31
0.86
0.44

657
C267T
267
0.85
0.2
0.62
0.56
0.27

658
C267P
267
0.81
0.28
0.53
0.54
0.21

659
Q268C
268
1.08
1.45
1.05
1.19
0.18

660
Q268G
268
0.71
1.13
0.53
0.79
0.25

661
Q268V
268
1.01
1.2
0.02
0.74
0.52

662
K269Y
269
0.2
0.42
1.02
0.55
0.35

663
N270G
270
0.65
0.82
1.63
1.03
0.43

664
N270E
270
0.74
0.9
0.15
0.6
0.32

665
K271W
271
0.63
0.8
0.78
0.73
0.07

666
K271H
271
0.61
0.98
0.42
0.67
0.23

667
K271C
271
−0.2
0.98
1
0.59
0.56

668
K271I
271
1.05
0.1
0.42
0.52
0.39

669
E272S
272
0.9
1.38
1.4
1.23
0.24

670
E272P
272
0.2
0.98
0.93
0.71
0.36

671
E272K
272
0.98
0.69
0.31
0.66
0.27

672
E272G
272
0.49
−0.12
1.12
0.5
0.51

673
E272Y
272
0.71
0.57
0.02
0.43
0.3

674
N273R
273
0.65
0.56
0.47
0.56
0.08

675
N273I
273
0.49
1.13
0.02
0.55
0.46

676
N273S
273
0.46
0.25
0.91
0.54
0.28

677
N275G
275
0.36
0.61
0.75
0.57
0.16

678
L276K
276
1.12
1.2
1.97
1.43
0.38

679
L276N
276
1.45
0.98
0.42
0.95
0.42

680
Y277P
277
1.3
1.2
1.48
1.33
0.12

681
Y277L
277
0.83
0.59
0.2
0.54
0.26

682
Y277M
277
0.8
0.08
0.39
0.42
0.29

683
K278W
278
1.74
0.98
0.42
1.05
0.54

684
K278A
278
0.67
1.07
0.71
0.82
0.18

685
K278P
278
0.45
0.98
0.71
0.71
0.21

686
K278S
278
0.85
0.69
0.22
0.59
0.26

687
K278F
278
−0.09
0.84
0.83
0.53
0.43

688
K278L
278
0.49
0.35
0.51
0.45
0.07

689
Q280M
280
0.89
0.5
0.58
0.66
0.17

690
Q280D
280
0.32
0.97
0.34
0.54
0.31

691
K281G
281
1.05
0.87
0.77
0.9
0.11

692
K281F
281
0.76
0.28
1.27
0.77
0.4

693
K281D
281
−0.61
0.48
2.2
0.69
1.16

694
L282S
282
−0.06
0.08
1.27
0.43
0.6

695
L282K
282
0.71
0.16
0.38
0.42
0.22

696
H283M
283
1.08
0.91
0.45
0.81
0.27

697
Q285M
285
1.44
1.52
2.1
1.68
0.29

698
Q285T
285
−0.76
0.71
1.95
0.64
1.11

699
I286C
286
1.45
0.28
0.71
0.81
0.48

700
C288G
288
0.57
0.69
0.57
0.61
0.06

701
C288K
288
0.56
0.42
0.57
0.52
0.07

702
I289W
289
0.87
0.66
0.33
0.62
0.22

703
A290K
290
0.59
0.54
0.37
0.5
0.09

704
A290I
290
0.28
0.69
0.35
0.44
0.18

705
A290N
290
0.68
0.59
0.01
0.43
0.29

706
A290R
290
0.15
0.38
0.72
0.42
0.23

707
D291T
291
1.21
0.46
1.87
1.18
0.57

708
D291M
291
0.85
0.63
0.76
0.75
0.09

709
D291N
291
0.63
0.68
0.77
0.7
0.06

710
D291F
291
0.49
0.75
0.8
0.68
0.14

711
D291P
291
0.48
0.36
1.08
0.64
0.32

712
D291L
291
0.63
0.62
0.29
0.51
0.16

713
D291W
291
0.52
0.69
0.3
0.5
0.16

714
T292H
292
0.42
0.46
0.54
0.47
0.05

715
S293K
293
0.89
0.64
0.61
0.71
0.12

716
S293W
293
0.19
0.46
1.02
0.56
0.35

717
S293H
293
0.38
0.66
0.52
0.52
0.12

718
Y294L
294
0.25
0.34
0.88
0.49
0.28

719
Y294M
294
0.74
0.37
0.31
0.47
0.19

720
Y294V
294
0.55
0.37
0.46
0.46
0.07

721
E295C
295
0.19
0.24
2.85
1.09
1.24

722
E295M
295
1.33
0.37
−0.01
0.56
0.56

723
E295T
295
0.64
0.58
0.36
0.52
0.12

724
E295R
295
0.49
0.55
0.46
0.5
0.04

725
V296Q
296
1.25
1.08
1.03
1.12
0.09

726
V296R
296
0.95
1.13
0.97
1.02
0.08

727
V296N
296
1.05
0.71
1.05
0.93
0.16

728
V296P
296
0.63
1.13
0.9
0.89
0.21

729
V296H
296
0.81
0.92
0.81
0.85
0.05

730
V296W
296
0.8
1.01
0.58
0.8
0.17

731
V296K
296
0.45
0.72
0.73
0.63
0.13

732
V296D
296
0.65
0.74
0.39
0.59
0.15

733
V296T
296
0.65
0.49
0.53
0.56
0.07

734
V296Y
296
0.7
0.54
0.35
0.53
0.14

735
V296A
296
0.47
0.44
0.64
0.52
0.09

736
V296M
296
0.76
0.45
0.31
0.5
0.19

737
V296G
296
0.38
0.43
0.48
0.43
0.04

738
P297F
297
1.34
1.36
1.49
1.4
0.07

739
P297I
297
1.46
0.88
0.93
1.09
0.26

740
P297C
297
1.26
1
0.7
0.99
0.23

741
P297V
297
0.85
0.76
1.06
0.89
0.12

742
P297W
297
0.83
0.91
0.91
0.88
0.04

743
P297H
297
0.76
0.73
0.52
0.67
0.11

744
P297N
297
0.77
0.64
0.53
0.64
0.1

745
P297M
297
0.69
0.5
0.6
0.59
0.08

746
P297K
297
0.54
0.61
0.56
0.57
0.03

747
P297Y
297
0.52
0.5
0.66
0.56
0.07

748
P297R
297
0.49
0.47
0.37
0.45
0.05

749
K299P
299
1.54
1.62
1.65
1.6
0.05

750
K299E
299
0.84
0.97
1.09
0.97
0.1

751
K299G
299
1.12
0.71
0.95
0.93
0.17

752
K299A
299
0.87
0.7
0.76
0.78
0.07

753
K299W
299
0.78
0.52
0.56
0.62
0.12

754
K299V
299
0.54
0.66
0.65
0.62
0.06

755
F300E
300
0.98
0.95
1.12
1.02
0.07

756
F300K
300
1.05
1.04
0.66
0.92
0.18

757
F300M
300
0.53
0.74
0.59
0.62
0.09

758
F300Q
300
0.8
0.71
0.22
0.58
0.26

759
F300D
300
0.71
0.34
0.37
0.47
0.17

760
E301G
301
1.94
1.88
2.02
1.95
0.06

761
E301W
301
1.42
1.71
1.45
1.53
0.13

762
E301P
301
1.58
1.48
0.99
1.35
0.26

763
E301Q
301
1.11
1.38
1.4
1.29
0.13

764
E301M
301
0.89
0.28
1.96
1.04
0.69

765
E301A
301
0.93
0.45
1.35
0.91
0.37

766
E301K
301
0.6
0.84
1.26
0.9
0.28

767
E301R
301
0.57
1.45
0.39
0.8
0.47

768
E301F
301
0.98
0.46
0.7
0.72
0.21

769
E301C
301
0.29
0.87
0.84
0.67
0.27

770
E301T
301
0.89
1.67
−0.68
0.62
0.98

771
S302I
302
2.18
2.07
2.31
2.19
0.1

772
S302D
302
1.49
2.23
1.86
1.86
0.3

773
S302V
302
1.67
1.84
1.66
1.72
0.08

774
S302G
302
1.44
1.14
1.6
1.39
0.19

775
S302E
302
1.58
0.84
1.57
1.33
0.35

776
S302M
302
1.58
1.67
0.48
1.24
0.54

777
S302A
302
0.85
1.18
0.89
0.97
0.15

778
S302W
302
1.08
0.77
0.97
0.94
0.13

779
S302N
302
0.64
0.82
1.01
0.82
0.15

780
S302C
302
0.48
0.97
0.52
0.66
0.22

781
S302H
302
0.89
0.28
0.7
0.62
0.25

782
D303C
303
3.03
3.65
3.23
3.3
0.26

783
D303M
303
3.14
3.09
2.97
3.07
0.07

784
D303F
303
2.59
2.84
2.71
2.71
0.1

785
D303Y
303
2.02
2.05
1.96
2.01
0.04

786
D303S
303
1.93
1.97
1.96
1.95
0.02

787
D303I
303
1.47
1.72
1.58
1.59
0.1

788
D303R
303
1.54
1.66
1.34
1.51
0.13

789
D303W
303
1.23
1.6
1.48
1.44
0.15

790
D303P
303
1.46
1.6
1.22
1.43
0.15

791
D303V
303
1.55
1.2
1.26
1.34
0.15

792
D303Q
303
1.14
1.32
1.35
1.27
0.09

793
D303G
303
1
1.27
1.18
1.15
0.11

794
D303H
303
1.09
1.13
1.18
1.13
0.04

795
D303K
303
0.76
1.09
1.12
0.99
0.16

796
D303N
303
0.63
0.72
0.76
0.7
0.05

797
E304T
304
0.69
0.61
0.76
0.69
0.06

798
E305W
305
1.98
1.85
2.18
2.01
0.14

799
E305C
305
1.39
1.75
1.9
1.68
0.22

800
E305F
305
1.61
1.56
1.64
1.6
0.03

801
E305S
305
1.51
1.46
1.69
1.55
0.1

802
E305Y
305
1.58
1.47
1.48
1.51
0.05

803
E305L
305
1.46
1.44
1.4
1.44
0.03

804
E305A
305
1.26
1.41
1.5
1.39
0.1

805
E305K
305
1.42
1.27
1.28
1.33
0.07

806
E3051
305
1.48
1.31
1.15
1.31
0.13

807
E305V
305
1.37
1.33
1.18
1.3
0.08

808
E305G
305
1.19
1.21
1.49
1.29
0.14

809
E305M
305
1.2
0.87
1.04
1.04
0.14

810
E305R
305
1.08
0.87
0.97
0.97
0.09

811
E305N
305
0.68
0.59
1.11
0.79
0.23

812
E305P
305
0.43
0.6
0.59
0.54
0.08

813
E305H
305
−0.08
1.04
0.43
0.46
0.46

814
V306W
306
2.13
1.38
1.39
1.63
0.35

815
V306Y
306
1.69
0.68
0.7
1.02
0.47

816
V306F
306
1.17
−0.13
0.38
0.47
0.53

817
V306C
306
0.23
0.72
0.32
0.43
0.21

818
Y307Q
307
1.88
2.18
2.02
2.03
0.12

819
Y307K
307
1.87
2.01
2.02
1.97
0.07

820
Y307E
307
1.83
2.04
1.94
1.94
0.09

821
Y307R
307
1.77
1.67
2.01
1.82
0.14

822
Y307G
307
1.81
1.72
1.86
1.8
0.06

823
Y307A
307
1.61
1.69
1.78
1.69
0.07

824
Y307T
307
1.62
1.72
1.68
1.68
0.04

825
Y307I
307
1.53
1.53
1.33
1.46
0.09

826
Y307L
307
1.26
1.1
1.25
1.2
0.08

827
Y307W
307
0.97
1.24
1.37
1.19
0.17

828
Y307V
307
1.08
1.17
1.15
1.13
0.04

829
Y307S
307
0.83
0.97
0.97
0.92
0.06

830
Y307P
307
0.92
0.52
0.71
0.72
0.16

831
Y307D
307
0.72
0.58
0.64
0.64
0.06

832
Y307N
307
0.4
0.54
0.48
0.47
0.06

833
Q308I
308
1.29
1.09
0.57
0.98
0.3

834
Q308W
308
0.53
0.54
0.51
0.53
0.01

835
S309M
309
0.57
0.95
0.81
0.78
0.15

836
S309L
309
0.44
0.68
0.49
0.54
0.1

837
S309K
309
0.52
0.36
0.61
0.5
0.1

838
S309V
309
0.5
0.22
0.59
0.43
0.16

839
V310Y
310
1.43
1.46
1.35
1.42
0.05

840
N311F
311
1.24
0.88
0.75
0.96
0.21

841
N311L
311
0.67
0.76
0.67
0.7
0.04

842
G312I
312
1.79
1.38
1.62
1.59
0.17

843
G312T
312
1.21
0.46
1.04
0.9
0.32

844
G312E
312
1.28
1.38
0.01
0.89
0.62

845
G312S
312
0.47
1.05
0.41
0.64
0.29

846
G312Q
312
0.87
0.97
0.01
0.62
0.43

847
G312A
312
0.6
0.53
0.52
0.55
0.03

848
G312K
312
−0.22
0.79
0.7
0.42
0.46

849
F313H
313
0.76
0.16
0.9
0.6
0.32

850
F313M
313
0.29
0.43
0.76
0.49
0.19

851
D315M
315
0.52
1.3
0.59
0.8
0.36

852
D315I
315
1.05
0.05
0.79
0.63
0.42

853
I317W
317
0.5
0.64
0.49
0.54
0.07

854
S318K
318
0.8
0.58
0.54
0.64
0.11

855
S318D
318
0.54
0.69
0.3
0.51
0.16

856
S319K
319
0.55
0.84
0.35
0.58
0.2

857
S319I
319
0.87
−0.13
0.79
0.51
0.46

858
S319N
319
−0.02
0.84
0.6
0.48
0.36

859
K320F
320
0.82
0.74
0.26
0.61
0.25

860
H321V
321
0.38
0.71
0.94
0.68
0.23

861
H321F
321
0.72
0.18
0.94
0.61
0.32

862
I322A
322
0.78
0.77
0.74
0.76
0.02

863
E324W
324
1.97
2.35
1.79
2.04
0.23

864
E324C
324
0.18
2.7
0.77
1.22
1.08

865
E324Y
324
0.78
0.43
0.77
0.66
0.17

866
E324M
324
0.87
0.49
0.22
0.53
0.26

867
E324T
324
0.18
0.35
0.77
0.43
0.25

868
R325K
325
0.17
0.78
0.69
0.55
0.27

869
R327V
327
0.76
0.4
0.55
0.57
0.15

870
R327M
327
0.58
0.5
0.56
0.55
0.03

871
R327K
327
0.18
0.85
0.29
0.44
0.3

872
D331M
331
0.84
0.78
0.58
0.73
0.11

873
D331C
331
0.92
0.68
0.34
0.65
0.24

874
Y333M
333
0.61
0.55
0.18
0.45
0.19

875
N334W
334
0.78
0.97
0.78
0.84
0.09

876
G335Q
335
0.62
0.49
0.28
0.46
0.14

877
Y336V
336
1.79
1.88
0.01
1.23
0.86

878
Y336H
336
1.43
0.68
0.92
1.01
0.31

879
Y336W
336
0.87
1.28
0.41
0.86
0.36

880
Y336M
336
0.47
0.97
0.29
0.58
0.29

881
N337G
337
0.89
0.51
0.33
0.58
0.23

882
D339Q
339
1.28
−0.42
0.7
0.52
0.7

883
D339M
339
0.87
0.68
0.01
0.52
0.37

884
D339S
339
−0.13
1.91
−0.4
0.46
1.03

885
K340E
340
0.69
1.12
1.47
1.09
0.32

886
K340H
340
1.28
0.97
0.01
0.75
0.54

887
K340A
340
0.87
0.27
1.1
0.75
0.35

888
Y342R
342
0.79
1.12
0.85
0.92
0.14

889
Y342M
342
0.99
−0.01
0.41
0.46
0.41

890
I343V
343
−0.51
0.97
0.85
0.44
0.67

891
V344E
344
1.28
0.97
1.1
1.12
0.13

892
K346C
346
0.18
1.88
0.7
0.92
0.71

893
F347R
347
0.87
1.66
1.61
1.38
0.36

894
F347G
347
0.18
0.27
0.92
0.46
0.33

895
Y348M
348
1.28
1.37
0
0.88
0.62

896
Y348V
348
0.18
0.27
1.39
0.61
0.55

897
E349T
349
2.26
0.96
1.1
1.44
0.58

898
E349Q
349
1.56
0.96
1.61
1.38
0.29

899
E349R
349
1.09
0.96
1.18
1.08
0.09

900
E349N
349
1.09
0.68
1.25
1.01
0.24

901
E349K
349
0.97
0.74
0.69
0.8
0.12

902
S350R
350
2.48
1.66
2.49
2.21
0.39

903
S350H
350
0.58
0.96
0
0.52
0.4

904
V351K
351
2.12
2.35
1.61
2.03
0.31

905
V351Y
351
1.28
1.19
1.71
1.39
0.23

906
V351F
351
1.43
0.27
1.1
0.93
0.49

907
V351D
351
1.79
0.56
0
0.78
0.75

908
S352M
352
0.18
1.19
1.39
0.92
0.53

909
Q353K
353
0.77
0.93
0.89
0.86
0.07

910
Q353M
353
0.58
0.43
1.38
0.8
0.42

911
Q353R
353
0.55
0.76
0.54
0.62
0.1

912
Q353N
353
0.17
0.54
0.57
0.43
0.18

913
T355M
355
0.83
0.47
0.4
0.57
0.19

914
Y356M
356
1.56
1.19
1.25
1.33
0.16

915
Y356V
356
0.96
0.86
0.79
0.87
0.07

916
Y356G
356
−0.11
1.37
0.69
0.65
0.61

917
Y356I
356
0.06
0.85
1.02
0.64
0.42

918
Y356C
356
0.51
0.27
0.95
0.58
0.28

919
Y356A
356
1.16
0.02
0.51
0.56
0.46

920
Y356E
356
−0.05
0.61
0.69
0.42
0.33

921
R357M
357
0.61
0.65
0.37
0.54
0.12

922
R357K
357
0.61
0.35
0.48
0.48
0.1

923
D358R
358
0.41
0.76
0.69
0.62
0.15

924
D358W
358
0.27
0.61
0.4
0.43
0.14

925
T361K
361
0.42
0.63
1.08
0.71
0.28

926
T364W
364
0.37
0.25
0.72
0.45
0.2

927
A365Y
365
1.04
0.8
0.14
0.66
0.38

928
A365N
365
0.62
0.4
0.64
0.56
0.11

929
E367P
367
0.76
0.68
0.52
0.65
0.1

930
E367M
367
0.58
0.41
0.29
0.43
0.12

931
I368P
368
1.4
0.72
0.91
1.01
0.29

932
I368Y
368
0.42
0.52
0.79
0.57
0.16

933
H369A
369
0.99
1.34
1.04
1.13
0.15

934
H369E
369
0.28
0.5
1.21
0.66
0.4

935
H369G
369
0.38
0.9
0.52
0.6
0.22

936
H369V
369
0.73
0.7
0.33
0.58
0.18

937
H369I
369
0.15
0.75
0.57
0.49
0.25

938
Y370A
370
0.71
0.99
0.53
0.74
0.19

939
Y370N
370
0.9
0.78
0.48
0.72
0.18

940
Y370G
370
0.78
0.39
0.81
0.66
0.19

941
Y370R
370
0.54
0.5
0.55
0.53
0.02

942
Y370P
370
0.47
0.49
0.58
0.51
0.05

943
N371C
371
1.1
1.2
0.76
1.02
0.19

944
N371G
371
0.61
0.77
0.07
0.49
0.3

945
N372F
372
1.16
0.83
0.85
0.94
0.15

946
N372A
372
0.52
0.72
0.47
0.57
0.11

947
N372W
372
0.52
0.62
0.19
0.44
0.18

948
I373D
373
0.6
0.8
0.39
0.59
0.17

949
L374H
374
0.89
0.68
1.1
0.89
0.17

950
L374Y
374
1.17
0.83
−0.03
0.66
0.51

951
L374E
374
0.61
0.65
0.37
0.55
0.12

952
L374N
374
0.55
0.24
0.53
0.44
0.14

953
P3751
375
0.45
0.58
0.45
0.49
0.06

954
P375Y
375
0.46
0.48
0.48
0.47
0.01

955
G376Q
376
0.63
1.05
1.12
0.93
0.22

956
N377V
377
0.94
0.89
0.45
0.76
0.22

957
N377L
377
0.55
0.42
0.46
0.48
0.05

958
N377P
377
0.81
0.28
0.15
0.42
0.28

959
G378N
378
0.41
0.9
0.49
0.6
0.21

960
G378W
378
0.4
0.45
0.59
0.48
0.08

961
K379F
379
0.65
0.75
0.59
0.66
0.07

962
K379A
379
0.52
0.53
0.48
0.51
0.02

963
K379R
379
0.17
0.65
0.66
0.49
0.23

964
K379G
379
0.58
0.53
0.21
0.44
0.16

965
K381L
381
0.56
0.63
0.51
0.57
0.05

966
K381W
381
0.57
0.82
0.23
0.54
0.24

967
K381P
381
0.45
0.24
0.56
0.42
0.13

968
D383M
383
0.61
0.56
0.5
0.56
0.05

969
D383P
383
0.5
0.51
0.33
0.45
0.08

970
K384D
384
0.15
0.86
0.78
0.59
0.32

971
K384P
384
0.58
0.41
0.56
0.52
0.08

972
K384G
384
0.62
0.61
0.25
0.49
0.17

973
V385C
385
0.41
0.61
0.5
0.51
0.08

974
V385Y
385
0.53
0.53
0.44
0.5
0.04

975
K386F
386
0.55
0.47
0.28
0.43
0.11

976
A388I
388
0.61
1.06
0.91
0.86
0.19

977
A388N
388
0.64
0.48
1.05
0.72
0.24

978
A388H
388
0.31
0.49
0.54
0.45
0.1

979
A388Y
388
0.57
0.65
0.08
0.43
0.25

980
V389E
389
0.82
0.22
0.41
0.48
0.25

981
K390C
390
0.19
0.6
0.54
0.44
0.18

982
N391W
391
0.7
0.71
0.93
0.78
0.11

983
N391Y
391
0.63
0.7
0.17
0.5
0.23

984
L393D
393
0.69
0.44
0.71
0.61
0.12

985
L393Y
393
0.41
0.54
0.5
0.48
0.05

986
Q394C
394
1.99
2.4
2.22
2.2
0.17

987
Q394M
394
0.79
0.78
0.8
0.79
0.01

988
Q394G
394
1.06
0.48
0.75
0.76
0.23

989
Q394W
394
0.8
0.29
0.96
0.68
0.29

990
Q394S
394
0.56
0.76
0.63
0.65
0.08

991
Q394A
394
0.59
0.68
0.61
0.63
0.04

992
Q394F
394
0.35
0.72
0.8
0.62
0.19

993
Q394V
394
0.58
0.52
0.7
0.6
0.07

994
Q394T
394
0.4
0.46
0.73
0.53
0.14

995
Q394I
394
0.52
0.32
0.41
0.42
0.08

996
K395V
395
0.41
0.57
0.6
0.53
0.08

997
K395I
395
0.3
0.52
0.62
0.48
0.13

998
K395F
395
0.5
0.44
0.33
0.42
0.07

999
I397Q
397
1.28
0.91
0.63
0.94
0.26

1000
I397C
397
0.6
0.58
0.55
0.58
0.02

1001
T398D
398
0.82
0.56
0.68
0.68
0.11

1002
T398F
398
0.35
0.68
0.67
0.57
0.15

1003
E399M
399
0.76
0.91
0.38
0.68
0.22

1004
I400E
400
1.1
0.21
−0.06
0.42
0.5

1005
E402H
402
0.86
0.12
0.49
0.49
0.3

1006
S405M
405
0.99
0.63
1.01
0.87
0.17

1007
S405K
405
0.46
0.64
0.48
0.53
0.08

1008
S405R
405
0.34
0.69
0.38
0.47
0.16

1009
N406A
406
0.53
0.61
0.63
0.59
0.04

1010
N406V
406
0.34
0.61
0.34
0.43
0.13

1011
Y407R
407
0.81
1.3
1.23
1.12
0.22

1012
Y407P
407
1.1
0.49
0.44
0.68
0.3

1013
Y407I
407
0.56
0.49
0.62
0.56
0.05

1014
K408I
408
0.91
0.72
0.92
0.85
0.09

1015
K408C
408
0.88
0.45
0.29
0.54
0.25

1016
K408D
408
0.24
0.65
0.49
0.46
0.17

1017
C410F
410
0.33
0.57
0.36
0.42
0.11

1018
S411N
411
1.31
1.39
1.03
1.24
0.15

1019
S411V
411
0.7
0.46
0.76
0.64
0.13

1020
D413W
413
0.78
0.32
0.68
0.59
0.2

1021
D413T
413
0.73
0.67
0.29
0.56
0.19

1022
D413R
413
0.68
0.29
0.52
0.5
0.16

1023
N414A
414
1.39
0.21
1.19
0.93
0.51

1024
N414D
414
0.59
0.83
0.63
0.68
0.11

1025
N414S
414
0.4
1.09
0.34
0.61
0.34

1026
N414L
414
0.23
1.03
0.24
0.5
0.38

1027
N414H
414
0.83
0.4
0.12
0.45
0.29

1028
I415E
415
0.43
0.9
−0.07
0.42
0.4

1029
I415C
415
−0.27
0.5
1.03
0.42
0.53

1030
K416P
416
0.71
0.86
0.5
0.69
0.15

1031
K416V
416
0.6
0.56
0.34
0.5
0.11

1032
K416W
416
0.74
0.5
0.22
0.49
0.21

1033
A417M
417
1.18
1.31
1.28
1.25
0.06

1034
A417C
417
0.89
0.43
1.52
0.95
0.44

1035
E418W
418
0.45
0.84
0.38
0.56
0.2

1036
T419G
419
0.83
0.21
1.72
0.92
0.62

1037
T419H
419
0.83
0.9
1.03
0.92
0.08

1038
T419Y
419
0.83
1.31
−0.76
0.46
0.88

1039
Y420V
420
0.83
1.12
1.32
1.09
0.2

1040
Y420R
420
0.39
0.82
1.16
0.79
0.32

1041
Y420K
420
0.62
0.48
0.99
0.7
0.22

1042
Y420Q
420
0.98
0.2
0.78
0.65
0.33

1043
I421F
421
1.23
2.15
1.72
1.7
0.38

1044
H422M
422
0.83
1.12
0.49
0.81
0.26

1045
H422C
422
0.13
1.59
0.33
0.68
0.64

1046
H422A
422
0.42
0.28
0.97
0.56
0.3

1047
H422F
422
0.64
0.49
0.44
0.52
0.09

1048
E423R
423
0.82
1.12
1.8
1.25
0.41

1049
E423T
423
0.54
1.3
−0.07
0.59
0.56

1050
E423S
423
0.82
0.2
0.62
0.55
0.26

1051
E423H
423
−0.09
0.79
0.88
0.53
0.44

1052
E423A
423
0.98
0.49
−0.07
0.47
0.43

1053
E423Y
423
0.13
1.01
0.15
0.43
0.41

1054
S425E
425
0.72
0.79
0.52
0.67
0.12

1055
S425K
425
0.25
1.17
0.41
0.61
0.4

1056
H426I
426
2
2.15
1.94
2.03
0.09

1057
H426G
426
1.31
0.89
0.74
0.98
0.24

1058
I427H
427
0.64
0.81
0.66
0.7
0.07

1059
L428T
428
0.13
0.89
0.62
0.55
0.32

1060
L428Y
428
0.38
0.45
0.51
0.45
0.05

1061
N429P
429
1.14
0.42
1.18
0.91
0.35

1062
N429A
429
0.35
1.45
0.33
0.71
0.52

1063
F431R
431
1.04
1.45
0.33
0.94
0.46

1064
F431A
431
0.41
0.71
0.91
0.68
0.2

1065
E432L
432
1.04
1.01
0.84
0.96
0.09

1066
E432R
432
0.82
0.42
0.74
0.66
0.17

1067
E432C
432
0.64
0.71
0.21
0.52
0.22

1068
E432T
432
1.11
0.2
0.21
0.51
0.42

1069
Q434F
434
1.22
1.81
1.87
1.63
0.29

1070
Q434W
434
1.73
0.89
2.01
1.54
0.47

1071
Q434S
434
1.04
0.89
0.84
0.93
0.08

1072
Q434I
434
0.82
0.2
1.72
0.91
0.62

1073
Q434L
434
0.89
0.74
0.55
0.72
0.14

1074
Q434P
434
−0.16
0.76
1.03
0.54
0.51

1075
Q434T
434
0.48
0.65
0.42
0.52
0.1

1076
E435A
435
0.97
0.89
0.62
0.83
0.15

1077
E435L
435
0.99
0.8
0.09
0.63
0.38

1078
E435Y
435
0.53
0.97
0.21
0.57
0.31

1079
L436G
436
2.18
2.4
2.21
2.26
0.1

1080
L436E
436
1.22
1.4
1.6
1.41
0.15

1081
L436T
436
0.82
0.99
0.8
0.87
0.08

1082
L436D
436
0.64
0.89
1.08
0.87
0.18

1083
L436A
436
1.35
0.2
1.03
0.86
0.48

1084
L436S
436
0.88
0.56
1.03
0.82
0.2

1085
L436W
436
0.82
0.47
0.96
0.75
0.21

1086
L436K
436
0.68
1.11
0.33
0.71
0.32

1087
L436P
436
1.29
0.38
0.4
0.69
0.42

1088
L436R
436
0.64
0.6
0.49
0.58
0.06

1089
K437P
437
0.64
1.3
1.03
0.99
0.27

1090
K437C
437
0.82
1.11
0.84
0.93
0.13

1091
K437H
437
1.3
0.2
0.74
0.75
0.45

1092
K437W
437
−0.28
1.04
0.77
0.51
0.57

1093
Y438R
438
1.85
2.08
2.07
2
0.11

1094
Y438A
438
0.89
0.98
1.13
1
0.1

1095
Y438W
438
1.01
1.06
0.92
1
0.06

1096
Y438P
438
0.95
0.55
0.63
0.71
0.17

1097
Y438K
438
0.56
0.77
0.72
0.68
0.09

1098
Y438I
438
0.76
0.71
0.51
0.66
0.11

1099
Y438G
438
0.55
0.25
0.58
0.46
0.15

1100
Y438L
438
0.4
0.52
0.43
0.45
0.05

1101
Y438E
438
0.35
0.51
0.46
0.44
0.07

1102
N439G
439
2.07
1.81
1.72
1.86
0.15

1103
N439P
439
1.3
1.45
1.31
1.35
0.07

1104
N439H
439
1.38
0.89
0.84
1.04
0.24

1105
N439A
439
0.91
−0.03
0.8
0.56
0.42

1106
N439S
439
0.65
0.2
0.39
0.42
0.19

1107
P440K
440
2.4
2.48
2.5
2.46
0.04

1108
P440R
440
1.82
1.89
1.94
1.89
0.05

1109
P440N
440
1.51
1.84
1.64
1.66
0.14

1110
P440D
440
1.42
2.2
1.22
1.61
0.42

1111
P440T
440
1.38
1.54
1.47
1.46
0.06

1112
P440Q
440
1.24
1.5
1.52
1.42
0.13

1113
P440E
440
1.26
1.42
1.46
1.38
0.08

1114
P440G
440
1.27
1.48
1.11
1.29
0.15

1115
P440L
440
0.9
1.28
1.21
1.13
0.16

1116
P440S
440
0.67
0.89
0.99
0.85
0.13

1117
P440A
440
0.77
0.52
0.68
0.66
0.1

1118
P440M
440
0.68
0.59
0.57
0.61
0.04

1119
E441H
441
1.42
1.81
1.39
1.54
0.19

1120
E441R
441
1.22
1.45
1.54
1.4
0.13

1121
E441K
441
1.12
1.03
1.31
1.15
0.12

1122
E441N
441
0.53
0.6
1.02
0.72
0.22

1123
E441F
441
0.63
0.71
0.62
0.65
0.04

1124
E441G
441
0.62
0.36
0.46
0.48
0.11

1125
I442P
442
1.33
1.93
1.47
1.57
0.26

1126
I442Y
442
0.81
0.84
1.23
0.96
0.19

1127
I442A
442
0.91
0.53
1.26
0.9
0.3

1128
I442G
442
1.14
0.08
0.03
0.42
0.51

1129
H443E
443
1.01
1.37
1.4
1.26
0.17

1130
H443F
443
1.51
0.6
1.32
1.14
0.39

1131
H443W
443
0.3
1.07
0.52
0.63
0.33

1132
H443A
443
0.41
1.18
0.22
0.6
0.42

1133
L444W
444
3.78
3.57
3.3
3.55
0.2

1134
L444A
444
2.07
2.42
2.4
2.29
0.16

1135
L444E
444
1.46
2.05
1.94
1.82
0.25

1136
L444Y
444
1.66
1.67
2.13
1.82
0.22

1137
L444K
444
1.73
1.81
1.63
1.73
0.07

1138
L444N
444
1.51
1.67
1.83
1.67
0.13

1139
L444T
444
1.45
1.46
1.57
1.49
0.05

1140
L444C
444
1.46
1.18
1.08
1.24
0.16

1141
L444D
444
0.53
1.5
1.6
1.21
0.49

1142
L444H
444
1.06
1.09
1.4
1.18
0.15

1143
L444G
444
1.04
0.71
1.27
1.01
0.23

1144
L444I
444
0.06
0.63
0.83
0.51
0.33

1145
V445K
445
2.65
2.76
2.89
2.77
0.1

1146
V445Q
445
2.02
1.05
1.82
1.63
0.42

1147
V445P
445
1.4
1.81
1.52
1.58
0.17

1148
V445E
445
1.4
1.33
1.35
1.36
0.03

1149
V445R
445
1.12
1.22
0.99
1.11
0.1

1150
V445C
445
0.57
0.89
0.29
0.58
0.25

1151
V445W
445
0.55
0.47
0.5
0.51
0.03

1152
E446W
446
0.82
1.3
1.88
1.33
0.43

1153
E446P
446
1.3
1.01
1.38
1.23
0.16

1154
E446T
446
0.53
0.61
0.34
0.49
0.11

1155
S447L
447
0.82
0.8
0.9
0.84
0.04

1156
S447P
447
0.8
0.9
0.75
0.82
0.06

1157
S447W
447
0.71
0.96
0.6
0.76
0.15

1158
S447F
447
0.4
0.66
0.63
0.56
0.11

1159
S447Q
447
0.67
0.54
0.4
0.54
0.11

1160
S447D
447
0.68
1.02
−0.2
0.5
0.51

1161
S447I
447
0.51
0.36
0.46
0.45
0.06

1162
S447Y
447
0.18
0.75
0.38
0.44
0.23

1163
E448W
448
1.22
1.38
1.25
1.29
0.07

1164
E448I
448
1.3
1.12
1.07
1.16
0.1

1165
E448Y
448
0.27
0.81
1.13
0.74
0.36

1166
E448H
448
0.78
0.73
0.53
0.68
0.11

1167
E448A
448
0.4
0.2
0.64
0.42
0.18

1168
L449D
449
0.51
0.52
0.22
0.42
0.14

1169
K450G
450
1.38
0.54
0.96
0.96
0.34

1170
K450S
450
0.43
0.64
0.39
0.49
0.11

1171
K450C
450
0.26
0.53
0.6
0.46
0.15

1172
K450V
450
0.71
0.39
0.2
0.43
0.21

1173
A451C
451
0.82
0.82
0.52
0.72
0.14

1174
A451M
451
0.68
0.28
0.82
0.59
0.23

1175
A451I
451
0.26
0.55
0.68
0.5
0.18

1176
S452P
452
1.25
1.42
1.44
1.37
0.09

1177
S452V
452
0.53
0.73
0.41
0.56
0.13

1178
S452G
452
0.45
0.75
0.3
0.5
0.18

1179
E453W
453
1.35
1.25
1.43
1.34
0.07

1180
E453F
453
0.4
0.47
0.43
0.43
0.03

1181
L454Y
454
0.65
0.59
0.54
0.6
0.05

1182
L454S
454
0.62
0.39
0.53
0.51
0.1

1183
L454R
454
0.61
0.57
0.26
0.48
0.15

1184
N456P
456
1.3
1.07
1.6
1.32
0.21

1185
N456M
456
1.1
1.51
1.09
1.24
0.19

1186
N456Q
456
0.12
0.5
1.04
0.55
0.38

1187
N456R
456
−0.28
0.56
1.12
0.46
0.58

1188
V457T
457
0.59
0.8
0.89
0.76
0.13

1189
V457Y
457
1.22
0.72
0.23
0.72
0.41

1190
V457E
457
0.82
0.5
−0.06
0.42
0.36

1191
V460N
460
1.14
0.61
1.27
1.01
0.28

1192
V460I
460
1.29
0.68
0.64
0.87
0.3

1193
V460K
460
0.82
0.97
0.64
0.81
0.14

1194
V460E
460
0.67
1.05
0.57
0.76
0.21

1195
V460P
460
0.63
0.9
0.45
0.66
0.18

1196
V460H
460
0.41
0.9
−0.06
0.42
0.39

1197
I461C
461
0.75
0.7
0.35
0.6
0.18

1198
I461Y
461
0.43
0.5
0.44
0.46
0.03

1199
N463A
463
0.4
0.34
0.59
0.44
0.11

1200
F465R
465
1.4
1.37
1.35
1.38
0.02

1201
F465Q
465
0.79
1.03
1.17
1
0.16

1202
F465K
465
0.83
1.05
1.06
0.98
0.1

1203
F465E
465
1.13
0.85
0.76
0.91
0.16

1204
F465T
465
0.89
0.73
0.81
0.81
0.07

1205
F465A
465
0.7
0.96
0.71
0.79
0.12

1206
F465V
465
0.73
0.94
0.51
0.73
0.18

1207
F465N
465
0.62
0.7
0.7
0.67
0.04

1208
F465G
465
0.55
0.44
0.62
0.54
0.07

1209
F465S
465
0.28
0.42
0.57
0.43
0.12

1210
H466K
466
0.65
0.87
0.58
0.7
0.13

1211
H466C
466
0.43
0.79
0.25
0.49
0.22

1212
S469K
469
0.98
1.19
1.14
1.1
0.09

1213
S469P
469
0.7
0.4
0.82
0.64
0.18

1214
S469L
469
0.6
0.57
0.66
0.61
0.03

1215
S469M
469
0.52
0.67
0.44
0.54
0.09

1216
S469V
469
0.44
0.45
0.6
0.5
0.07

1217
S469R
469
0.39
0.5
0.42
0.44
0.05

1218
V470K
470
1.04
1.06
1.11
1.07
0.03

1219
V470N
470
0.65
0.44
0.67
0.59
0.11

1220
V470Y
470
0.65
0.48
0.59
0.57
0.07

1221
M472G
472
0.35
0.88
0.57
0.6
0.22

1222
T473R
473
0.18
0.89
0.46
0.51
0.3

1223
T473G
473
−0.23
1.62
0.05
0.48
0.82

1224
T473F
473
0.41
0.21
0.65
0.42
0.18

1225
E474P
474
0.78
1.04
0.87
0.9
0.11

1226
L476R
476
1.05
1.29
1.13
1.16
0.1

1227
L476K
476
0.61
0.49
0.75
0.62
0.11

1228
L476H
476
0.46
0.74
0.49
0.56
0.13

1229
L476T
476
0.39
0.58
0.49
0.48
0.08

1230
L476G
476
0.43
0.56
0.42
0.47
0.06

1231
L476N
476
0.47
0.5
0.36
0.44
0.06

1232
V477Q
477
1.14
1.43
1.34
1.3
0.12

1233
V477T
477
1.12
1.04
1.54
1.23
0.22

1234
V477W
477
1.3
1.87
−0.73
0.81
1.12

1235
V477E
477
1.07
0.29
0.67
0.68
0.32

1236
V477D
477
0.62
0.75
0.15
0.51
0.26

1237
V477M
477
0.53
0.42
0.44
0.46
0.05

1238
K479A
479
0.13
1.2
1.16
0.83
0.5

1239
K479H
479
1.04
−0.48
1.06
0.54
0.72

1240
K479D
479
0.44
0.59
0.45
0.49
0.07

1241
D480Q
480
0.69
0.33
0.88
0.63
0.23

1242
D480P
480
0.34
0.29
0.65
0.43
0.16

1243
N481A
481
2.07
0.91
1.06
1.35
0.52

1244
N481D
481
0.46
0.91
0.3
0.56
0.26

1245
N482A
482
1.79
1.66
2.07
1.84
0.17

1246
N482H
482
1.04
0.22
1.47
0.91
0.52

1247
N482Q
482
0.69
0.44
0.97
0.7
0.22

1248
N482D
482
0.13
0.91
0.25
0.43
0.34

1249
N482S
482
0.38
0.47
0.41
0.42
0.04

1250
F483W
483
0.64
0.77
0.35
0.58
0.18

1251
F483A
483
0.39
0.4
0.8
0.53
0.19

1252
Y484G
484
1.23
0.22
1.35
0.93
0.51

1253
Y484V
484
0.69
0.44
0.53
0.55
0.1

1254
Y484R
484
0.82
0.34
0.45
0.54
0.21

1255
A485Q
485
0.62
0.45
1.15
0.74
0.3

1256
A485K
485
0.63
1.08
0.37
0.69
0.29

1257
A485R
485
0.32
0.76
0.99
0.69
0.28

1258
A485L
485
0.73
0.67
0.45
0.62
0.12

1259
A485H
485
0.38
0.84
0.42
0.55
0.21

1260
A485S
485
0.7
0.4
0.46
0.52
0.13

1261
E486L
486
1.89
1.95
1.19
1.68
0.34

1262
E486R
486
1.23
1.61
1.22
1.35
0.18

1263
E486S
486
0.82
1.76
1.44
1.34
0.39

1264
E486Y
486
1.52
1.32
1.15
1.33
0.15

1265
E486F
486
0.69
1.14
0.53
0.79
0.26

1266
E486A
486
1.36
0.41
0.56
0.77
0.42

1267
E486G
486
0.54
0.52
0.19
0.42
0.16

1268
E488Q
488
0.42
1.43
0.82
0.89
0.41

1269
E488T
488
0.54
0.92
0.78
0.75
0.16

1270
E489H
489
2.08
1.14
1.23
1.48
0.42

1271
E489C
489
0.72
1.18
1.2
1.03
0.22

1272
E489T
489
0.86
1.06
0.53
0.82
0.22

1273
E489M
489
0.36
0.63
0.86
0.61
0.2

1274
E489S
489
0.33
0.64
0.53
0.5
0.13

1275
E489W
489
0.38
0.23
0.7
0.43
0.2

1276
I490A
490
0.47
0.56
1.07
0.7
0.27

1277
I490E
490
0.37
0.65
0.54
0.52
0.12

1278
Y491E
491
1.23
0.92
1.23
1.13
0.15

1279
Y491Q
491
0.83
0.22
0.89
0.65
0.3

1280
D492I
492
1.67
1.69
1.52
1.63
0.08

1281
D492L
492
0.4
0.76
0.56
0.57
0.15

1282
D492K
492
0.29
0.04
1.36
0.56
0.57

1283
E493C
493
0.65
0.59
0.08
0.44
0.25

1284
E493R
493
0.43
0.35
0.51
0.43
0.07

1285
Y495V
495
0.49
0.78
0.77
0.68
0.14

1286
P496W
496
0.14
1.62
2.28
1.35
0.9

1287
P496E
496
1.23
0.92
1.23
1.13
0.15

1288
P496C
496
0.98
1.08
0.96
1.01
0.05

1289
P496V
496
0.98
0.74
1.28
1
0.22

1290
P496L
496
0.7
0.96
0.65
0.77
0.14

1291
P496K
496
0.95
0.45
0.9
0.77
0.22

1292
P496S
496
0.57
0.34
0.4
0.43
0.1

1293
P496I
496
0.83
−0.46
0.9
0.42
0.63

1294
V497W
497
0.17
0.88
1.05
0.7
0.38

1295
I498R
498
1.21
1.15
0.91
1.09
0.13

1296
I498K
498
0.86
0.75
0.64
0.75
0.09

1297
I498P
498
0.59
0.63
0.71
0.65
0.05

1298
I498G
498
0.28
0.66
0.63
0.52
0.17

1299
S499M
499
0.98
0.74
0.67
0.8
0.13

1300
S499C
499
1.23
1.04
−0.31
0.65
0.68

1301
S499G
499
0.83
0.23
0.67
0.58
0.26

1302
L500S
500
0.27
0.95
0.17
0.47
0.34

1303
L503N
503
0.82
2.3
2.54
1.89
0.76

1304
L503P
503
1.23
1.14
0.99
1.12
0.1

1305
L503H
503
0.64
1.2
0.59
0.81
0.28

1306
L503I
503
1.31
0.45
0.67
0.81
0.37

1307
L503G
503
0.69
0.78
0.67
0.72
0.05

1308
L503R
503
0.31
0.69
0.94
0.65
0.26

1309
L503D
503
0.42
1.2
0.27
0.63
0.41

1310
L503K
503
0.31
0.69
0.86
0.62
0.23

1311
L503E
503
0.54
0.22
1.08
0.61
0.35

1312
L503W
503
0.42
1.07
−0.02
0.49
0.45

1313
V504A
504
1.26
1.45
1.64
1.45
0.16

1314
V504T
504
0.96
1.09
1.21
1.08
0.1

1315
V504G
504
0.75
0.61
0.74
0.7
0.06

1316
V504C
504
0.21
0.67
0.55
0.48
0.19

1317
Y507R
507
2.44
2.56
2.77
2.59
0.13

1318
Y507K
507
2.45
2.47
2.36
2.43
0.05

1319
Y507W
507
1.96
2.04
2.26
2.09
0.13

1320
Y507S
507
1.92
1.99
2.05
1.99
0.05

1321
Y507A
507
1.79
1.86
1.78
1.81
0.04

1322
Y507T
507
1.5
1.73
1.79
1.67
0.13

1323
Y507Q
507
1.6
1.52
1.87
1.67
0.15

1324
Y507C
507
1.41
1.28
1.53
1.41
0.1

1325
Y507V
507
0.99
1.09
1.21
1.1
0.09

1326
Y507E
507
1.23
0.63
1.43
1.1
0.34

1327
Y507G
507
1.17
0.77
0.95
0.96
0.16

1328
Y507L
507
0.99
0.86
0.7
0.85
0.12

1329
Y507I
507
1
0.66
0.85
0.84
0.14

1330
T509L
509
0.62
1.01
1.1
0.91
0.21

1331
T509V
509
0.31
1.29
0.38
0.66
0.44

1332
T509M
509
1.01
0.81
0.07
0.63
0.4

1333
Q510F
510
2.46
2.65
2.77
2.63
0.13

1334
Q510G
510
2.19
2.39
2.52
2.37
0.14

1335
Q510Y
510
2.22
2.26
2.36
2.28
0.06

1336
Q510S
510
2.12
2.37
2.3
2.26
0.1

1337
Q510C
510
2.08
2.16
2.18
2.14
0.05

1338
Q510V
510
2.01
2.17
2.21
2.13
0.09

1339
Q510W
510
1.95
2.19
2.16
2.1
0.11

1340
Q510N
510
1.91
1.85
2.22
1.99
0.16

1341
Q510A
510
1.58
1.81
1.92
1.77
0.14

1342
Q510D
510
1.56
1.97
1.46
1.66
0.22

1343
Q510I
510
1.77
1.46
1.75
1.66
0.14

1344
Q510H
510
1.04
1.19
1.13
1.12
0.06

1345
Q510K
510
0.83
0.99
0.94
0.92
0.07

1346
Q510M
510
0.75
0.67
0.63
0.68
0.05

1347
Q510E
510
0.64
0.69
0.61
0.65
0.03

1348
Q510P
510
0.68
0.58
0.59
0.62
0.04

1349
Q510T
510
0.48
0.72
0.63
0.61
0.1

1350
Q510R
510
0.35
0.47
0.5
0.44
0.07

1351
K511Y
511
0.96
1.37
0.33
0.88
0.43

1352
K511G
511
1.45
0.85
0.29
0.87
0.47

1353
K511S
511
0.85
0.75
0.79
0.79
0.04

1354
K511F
511
0.99
0.78
0.38
0.72
0.25

1355
K511V
511
0.45
0.92
0.63
0.67
0.19

1356
P512N
512
2.51
2.77
2.8
2.69
0.13

1357
P512F
512
2.04
2.32
2.29
2.22
0.13

1358
P512Y
512
1.9
2.03
2.11
2.02
0.09

1359
P512C
512
1.96
1.98
2.03
1.99
0.03

1360
P512G
512
1.86
1.91
1.96
1.91
0.04

1361
P512D
512
1.74
1.88
1.85
1.82
0.06

1362
P512I
512
1.45
1.69
1.73
1.62
0.12

1363
P512H
512
1.53
1.61
1.58
1.57
0.03

1364
P512V
512
1.46
1.48
1.69
1.54
0.11

1365
P512W
512
1.08
1.29
1.4
1.26
0.13

1366
P512S
512
0.97
1.09
1.06
1.04
0.05

1367
P512E
512
0.88
0.74
0.81
0.81
0.06

1368
P512T
512
0.5
0.69
0.49
0.56
0.09

1369
P512R
512
0.47
0.48
0.43
0.46
0.02

1370
Y513G
513
3.41
3.77
3.79
3.66
0.18

1371
Y513Q
513
3.29
3.52
3.71
3.51
0.17

1372
Y513R
513
3.28
3.4
3.53
3.41
0.1

1373
Y513K
513
3.27
3.36
3.45
3.36
0.07

1374
Y513N
513
3.11
3.13
3.18
3.14
0.03

1375
Y513I
513
2.88
2.96
2.88
2.91
0.04

1376
Y513A
513
2.6
2.65
2.8
2.68
0.08

1377
Y513S
513
2.47
2.62
2.74
2.61
0.11

1378
Y513E
513
2.53
2.59
2.68
2.6
0.06

1379
Y513V
513
2.5
2.51
2.63
2.55
0.06

1380
Y513L
513
2.35
2.58
2.54
2.49
0.1

1381
Y513D
513
2.29
2.59
2.59
2.49
0.14

1382
Y513C
513
2.31
2.57
2.47
2.45
0.11

1383
Y513M
513
2.16
2.37
2.27
2.27
0.08

1384
Y513T
513
2.21
2.23
2.33
2.26
0.05

1385
Y513H
513
1.5
2.33
2.47
2.1
0.43

1386
Y513P
513
0.85
1.09
1.15
1.03
0.13

1387
S514P
514
3.68
3.78
3.95
3.8
0.11

1388
S514W
514
1.97
2.14
1.99
2.03
0.08

1389
S514G
514
0.96
1.29
1.4
1.22
0.19

1390
S514H
514
0.89
0.72
1
0.87
0.12

1391
S514F
514
1.16
0.79
0.66
0.87
0.21

1392
S514D
514
0.73
1.22
0.64
0.86
0.25

1393
S514R
514
0.97
0.56
0.89
0.81
0.18

1394
S514A
514
0.73
0.94
0.69
0.79
0.11

1395
S514K
514
0.69
0.76
0.61
0.68
0.06

1396
S514N
514
0.69
0.66
0.24
0.53
0.21

1397
S514L
514
0.6
0.47
0.46
0.51
0.06

1398
T515G
515
2.89
3.08
2.85
2.94
0.1

1399
T515C
515
1.25
1.53
1.85
1.54
0.24

1400
T515P
515
1.31
1.48
1.49
1.43
0.09

1401
T515A
515
0.83
0.95
1.01
0.93
0.08

1402
T515L
515
0.51
0.71
0.57
0.6
0.08

1403
T515M
515
0.66
0.38
0.66
0.57
0.13

1404
T515S
515
0.4
0.58
0.44
0.47
0.08

1405
T515Y
515
0.49
0.56
0.3
0.45
0.11

1406
K516P
516
2.25
2.32
2.58
2.38
0.14

1407
K516S
516
1.2
1.23
1.25
1.23
0.02

1408
K516R
516
0.94
0.89
0.93
0.92
0.02

1409
K516A
516
0.63
0.72
0.79
0.71
0.07

1410
K516T
516
0.43
0.54
0.55
0.51
0.05

1411
K517P
517
3.68
3.83
3.89
3.8
0.09

1412
K517D
517
1.33
1.61
1.08
1.34
0.22

1413
K517L
517
0.96
1.32
1.27
1.18
0.16

1414
K517G
517
1.12
0.61
1.13
0.95
0.24

1415
K517A
517
0.42
1.07
0.67
0.72
0.27

1416
I518P
518
3.56
3.55
3.85
3.65
0.14

1417
I518T
518
2.35
2.64
2.51
2.5
0.12

1418
I518S
518
1.38
1.39
1.52
1.43
0.07

1419
I518Q
518
1.23
1.48
1.43
1.38
0.11

1420
I518E
518
0.85
0.91
1.13
0.96
0.12

1421
I518L
518
0.54
0.5
0.82
0.62
0.14

1422
I518F
518
0.38
0.84
0.4
0.54
0.21

1423
L520M
520
3.57
3.87
4.02
3.82
0.19

1424
L520N
520
3.29
3.55
3.52
3.45
0.12

1425
L520Q
520
2.85
3.24
3.31
3.13
0.2

1426
L520H
520
2.54
2.25
2.58
2.46
0.14

1427
L520S
520
2.05
2.41
2.4
2.29
0.17

1428
L520G
520
1.86
1.7
1.72
1.76
0.07

1429
L520A
520
0.64
0.29
0.45
0.46
0.14

1430
N521F
521
1.74
1.07
0.82
1.21
0.39

1431
N521P
521
1.31
0.44
1.36
1.04
0.42

1432
N521R
521
0.82
0.56
0.93
0.77
0.16

1433
N521L
521
−0.16
1.23
0.79
0.62
0.58

1434
N521E
521
0.47
0.4
0.76
0.54
0.16

1435
F522A
522
2.92
3.19
3.15
3.09
0.12

1436
F522G
522
2.43
2.47
2.5
2.46
0.03

1437
F522M
522
1.06
1.02
0.63
0.91
0.2

1438
F522C
522
0.85
0.79
0.84
0.83
0.02

1439
G523Q
523
1.92
0.92
0.67
1.17
0.54

1440
G523A
523
1.52
0.92
1.07
1.17
0.25

1441
A528I
528
0.53
0.92
−0.03
0.47
0.39

1442
Y537F
537
0.17
0.79
0.67
0.54
0.27

1443
S538C
538
0.41
0.21
0.69
0.44
0.2

1444
S538N
538
0.46
0.41
0.4
0.42
0.03

1445
A541N
541
2.61
2.6
2.81
2.67
0.1

1446
A541I
541
2.27
2.51
2.54
2.44
0.12

1447
A541V
541
1.66
1.94
1.82
1.81
0.11

1448
A541M
541
1.68
0.62
1.46
1.25
0.46

1449
A541L
541
0.53
1.06
0.73
0.77
0.22

1450
A541T
541
0.65
0.78
0.86
0.77
0.09

1451
A541Q
541
0.41
0.91
0.87
0.73
0.23

1452
I543H
543
1.73
1.94
1.63
1.77
0.13

1453
I543W
543
2.2
1.12
1.2
1.51
0.49

1454
I543F
543
1.22
1.73
1.41
1.45
0.21

1455
I543G
543
1.6
0.9
0.54
1.01
0.44

1456
I543T
543
0.81
1.33
0.84
1
0.24

1457
I543A
543
0.71
0.39
0.83
0.64
0.18

1458
I543C
543
0.97
0.5
−0.05
0.47
0.42

1459
I543S
543
0.41
0.47
0.45
0.45
0.02

1460
I543Q
543
0.81
0.21
0.24
0.42
0.28

1461
L544Y
544
1.02
1.42
1.36
1.27
0.17

1462
L544W
544
1.04
0.43
0.64
0.7
0.25

1463
M545E
545
0.52
0.21
1.45
0.73
0.53

1464
M545Q
545
−0.22
1.21
0.3
0.43
0.59

1465
R546F
546
0.96
1.19
1.55
1.24
0.24

1466
L549D
549
1.03
0.61
−0.06
0.53
0.45

1467
L552E
552
2.72
2.69
2.68
2.69
0.02

1468
L552Q
552
1.26
1.59
2.1
1.65
0.35

1469
L552K
552
0.81
0.61
0.92
0.78
0.13

1470
G553M
553
2
2.06
2
2.02
0.03

1471
G553L
553
1.77
1.79
1.83
1.8
0.03

1472
G553T
553
1.69
1.73
1.82
1.75
0.06

1473
G553I
553
1.1
1.39
1.28
1.26
0.12

1474
G553V
553
0.81
0.96
0.92
0.9
0.06

1475
G553S
553
0.66
0.73
0.76
0.72
0.04

1476
G553A
553
0.56
0.72
0.6
0.63
0.07

1477
I554C
554
0.69
0.94
1.23
0.95
0.22

1478
F555M
555
0.84
0.74
0.43
0.67
0.18

1479
N556E
556
3.02
3.2
2.94
3.05
0.11

1480
N556Q
556
1.85
1.99
2.42
2.09
0.24

1481
N556C
556
1.65
1.81
2.27
1.91
0.26

1482
N556M
556
1.62
1.73
1.85
1.74
0.09

1483
N556R
556
1.66
1.62
1.69
1.66
0.03

1484
N556A
556
1.36
1.66
1.82
1.61
0.19

1485
N556V
556
1.43
1.38
1.57
1.46
0.08

1486
N556F
556
0.93
1.07
1.14
1.05
0.09

1487
N556W
556
0.54
0.66
0.62
0.61
0.05

1488
N556K
556
0.4
0.44
0.45
0.43
0.02

1489
A557S
557
1.62
1.74
1.6
1.66
0.06

1490
A557K
557
1.21
1.23
0.96
1.13
0.12

1491
K558H
558
1.98
2.28
2.18
2.15
0.12

1492
K558M
558
2.06
1.99
0.62
1.56
0.66

1493
K558S
558
1.37
1.7
1.54
1.54
0.14

1494
K558F
558
1.41
0.71
1.66
1.26
0.4

1495
K558E
558
0.81
1.3
0.62
0.91
0.29

1496
K558T
558
0.81
1.12
0.74
0.89
0.16

1497
N559G
559
1.09
1.54
1.72
1.45
0.26

1498
N559E
559
1.21
1.58
1.18
1.33
0.18

1499
N559A
559
0.81
1.14
1.16
1.04
0.16

1500
D562I
562
1.59
1.52
1.75
1.62
0.09

1501
D562L
562
1.51
1.54
1.6
1.55
0.04

1502
D562F
562
1.43
1.32
1.34
1.36
0.05

1503
D562T
562
1.02
1.43
1.42
1.29
0.19

1504
D562Q
562
1.21
1.19
1.21
1.2
0.01

1505
D562M
562
1.01
1.13
1.18
1.11
0.08

1506
D562W
562
0.68
1
0.9
0.86
0.14

1507
D562C
562
0.59
0.93
1.05
0.86
0.19

1508
D562V
562
0.68
0.83
0.86
0.79
0.08

1509
D562K
562
0.31
0.57
0.66
0.51
0.15

1510
K564A
564
1.82
1.29
1.31
1.47
0.24

1511
K564G
564
0.88
1.45
0.08
0.8
0.56

1512
K564D
564
0.62
0.71
1.03
0.78
0.17

1513
I565R
565
0.29
−0.32
1.93
0.63
0.95

1514
I565F
565
0.45
0.66
0.26
0.46
0.16

1515
I566A
566
0.8
1.15
0.71
0.89
0.19

1516
I566W
566
1.21
−0.21
0.44
0.48
0.58

1517
E567W
567
1.09
1.49
1.23
1.27
0.17

1518
E567Q
567
0.7
0.98
1.09
0.92
0.17

1519
E567L
567
0.38
0.3
0.59
0.42
0.12

1520
N569P
569
0.84
0.95
0.8
0.86
0.06

1521
N569W
569
0.56
0.65
0.62
0.61
0.03

1522
T570F
570
1.72
1.8
1.31
1.61
0.21

1523
T570D
570
0.8
0.6
0.33
0.58
0.19

1524
S571W
571
1.21
1.8
1.72
1.58
0.26

1525
S571Q
571
1.9
1.29
1.31
1.5
0.28

1526
S571Y
571
1.03
1.8
1.43
1.42
0.32

1527
S571M
571
−0.58
0.89
1.18
0.5
0.77

1528
E572M
572
1.5
0.2
0.62
0.77
0.54

1529
K574F
574
2.41
2.84
1.54
2.26
0.54

1530
K574V
574
1.41
0.71
−0.48
0.55
0.78

1531
K574I
574
0.8
0.6
−0.07
0.44
0.38

1532
G575D
575
0.8
1.3
−0.07
0.68
0.57

1533
G575W
575
0.4
0.71
0.21
0.44
0.21

1534
Y577P
577
1.03
0.6
1.31
0.98
0.29

1535
Y577M
577
−0.18
1.01
1.31
0.71
0.64

1536
K578F
578
1.29
0.42
1.02
0.91
0.36

1537
K578C
578
1.21
0.2
0.62
0.67
0.41

1538
K578A
578
0.68
0.4
0.82
0.63
0.18

1539
M580Q
580
0.44
1.15
0.88
0.82
0.29

1540
I581H
58
0.55
0.3
0.88
0.58
0.24

1541
N583V
583
0.67
1.13
1.07
0.96
0.2

1542
N583T
583
0.66
0.93
0.53
0.71
0.16

1543
N583G
583
0.42
0.89
0.55
0.62
0.2

1544
N583A
583
0.65
0.57
0.56
0.59
0.04

1545
N583I
583
0.69
0.62
0.39
0.57
0.13

1546
N583S
583
0.45
0.72
0.42
0.53
0.13

1547
L585K
585
0.51
0.5
0.87
0.63
0.17

1548
P586F
586
1.71
0.89
1.02
1.21
0.36

1549
G587C
587
1.2
0.89
1.31
1.13
0.18

1550
G587H
587
0.8
0.6
0.84
0.75
0.1

1551
G602A
602
0.8
1.48
1.21
1.16
0.28

1552
E604I
604
0.36
0.89
0.55
0.6
0.22

1553
E604T
604
0.51
0.82
0.41
0.58
0.17

1554
T605C
605
0.47
1.08
1.16
0.9
0.31

1555
T605G
605
0.8
0.89
0.74
0.81
0.06

1556
K607C
607
0.93
0.57
0.63
0.71
0.16

1557
S609E
609
0.45
1.03
0.73
0.74
0.24

1558
S609C
609
0.64
0.84
0.68
0.72
0.09

1559
S609K
609
0.67
0.35
0.85
0.63
0.21

1560
A610N
610
0.8
1.59
1.32
1.24
0.33

1561
A610I
610
0.9
0.67
1.04
0.87
0.15

1562
A610H
610
0.9
0.38
0.63
0.64
0.21

1563
A610Y
610
−0.99
1.3
1.14
0.49
1.04

1564
L613G
613
0.99
1.08
1.17
1.08
0.07

1565
E614I
614
0.52
0.21
1.65
0.79
0.62

1566
K617V
617
0.39
0.45
0.54
0.46
0.06

1567
K617C
617
0.4
0.42
0.46
0.43
0.02

1568
K617Q
617
0.5
0.51
0.26
0.43
0.12

1569
Q618D
618
0.8
0.59
0.48
0.62
0.13

1570
N619R
619
0.26
0.59
0.88
0.58
0.26

1571
N619C
619
0.44
0.53
0.58
0.52
0.06

1572
K620F
620
0.55
0.21
0.52
0.43
0.15

1573
S624W
624
0.52
1.13
0.11
0.59
0.42

1574
S625N
625
0.44
0.48
0.44
0.46
0.02

1575
K626I
626
1.04
0.44
0.97
0.82
0.27

1576
K626V
626
0.81
0.12
0.82
0.58
0.33

1577
K626Y
626
0.52
0.37
0.73
0.54
0.15

1578
H634K
634
0.08
1.08
0.55
0.57
0.41

1579
L636H
636
0.82
0.22
0.66
0.57
0.25

1580
D638I
638
1.51
0.22
−0.03
0.57
0.68

1581
D638C
638
0.85
0.24
0.38
0.49
0.26

1582
D638S
638
0.41
0.04
0.88
0.45
0.35

1583
Y639N
639
0.82
0.92
1.07
0.93
0.1

1584
Y639M
639
0.82
1.32
−0.03
0.7
0.56

1585
Y639G
639
0.82
0.78
0.19
0.6
0.29

1586
Y639T
639
0.64
0.04
0.89
0.52
0.36

1587
Y639H
639
0.13
1.32
−0.03
0.47
0.6

1588
N642A
642
0.28
0.83
1.12
0.74
0.35

1589
N642M
642
0.32
0.8
0.47
0.53
0.2

1590
C643T
643
0.13
1.24
0.66
0.68
0.45

1591
C643F
643
0.53
0.23
0.53
0.43
0.14

1592
I644Y
644
0.66
0.33
0.61
0.53
0.14

1593
I644A
644
0.44
0.34
0.72
0.5
0.16

1594
A645M
645
0.73
0.74
0.38
0.62
0.17

1595
A645N
645
0.47
0.52
0.58
0.52
0.04

1596
A645Q
645
0.57
0.45
0.42
0.48
0.07

1597
P648C
648
1.28
0.84
0.82
0.98
0.21

1598
P648M
648
0.13
1.02
0.76
0.64
0.38

1599
P648I
648
0.67
0.24
0.57
0.49
0.19

1600
F653R
653
0.95
0.56
1.02
0.85
0.2

1601
G654P
654
0.61
0.44
0.75
0.6
0.13

1602
F655H
655
0.45
0.72
0.99
0.72
0.22

1603
D656K
656
0.88
0.87
0.55
0.77
0.15

1604
D656G
656
0.5
0.78
0.73
0.67
0.12

1605
S658D
658
0.3
0.78
0.41
0.5
0.21

1606
D659K
659
0.45
0.35
0.61
0.47
0.1

1607
S661G
661
0.88
0.97
0.47
0.77
0.22

1608
S661Q
661
0.72
0.37
0.39
0.49
0.16

1609
S661M
661
0.06
0.93
0.44
0.48
0.36

1610
E664R
664
0.45
0.93
0.43
0.61
0.23

1611
E664F
664
0.61
0.71
0.46
0.59
0.1

1612
E664S
664
0.27
0.42
0.61
0.43
0.14

1613
S667R
667
1.52
0.53
0.97
1.01
0.41

1614
G668Q
668
2.7
0.56
−0.3
0.99
1.26

1615
G668M
668
0.25
0.36
0.68
0.43
0.18

1616
F669P
669
1.46
1.42
1.43
1.44
0.02

1617
F669R
669
1.62
−0.27
0.17
0.51
0.81

1618
L675C
675
1.34
1.23
1.78
1.45
0.24

1619
Q676V
676
1.02
1.2
1.06
1.1
0.08

1620
Q676T
676
0.88
1.2
0.66
0.91
0.22

1621
Q676S
676
0.6
1.13
0.89
0.87
0.22

1622
Q676W
676
0.14
0.5
0.93
0.52
0.32

1623
Y678F
678
1.28
1.81
1.71
1.6
0.23

1624
Y678H
678
0.67
1.05
1.32
1.01
0.27

1625
I680A
680
1.63
1.34
1.34
1.44
0.14

1626
I680V
680
1.2
1.4
1.26
1.29
0.08

1627
I680Q
680
1.29
1.1
1.36
1.25
0.11

1628
I680S
680
1.03
1.04
1.07
1.05
0.02

1629
I680N
680
1.06
1.07
0.79
0.97
0.13

1630
I680H
680
0.92
1.17
0.78
0.96
0.16

1631
I680F
680
0.52
0.58
0.38
0.49
0.09

1632
D681I
681
1.11
1.21
1.16
1.16
0.04

1633
D681W
681
1.11
0.9
1.01
1.01
0.08

1634
D681M
681
0.53
0.68
0.79
0.67
0.11

1635
D681Q
681
0.48
0.8
0.65
0.64
0.13

1636
D681T
681
0.59
0.62
0.7
0.64
0.05

1637
D681R
681
0.54
0.49
0.58
0.54
0.04

1638
D681K
681
0.45
0.42
0.47
0.45
0.02

1639
W682D
682
1.37
1.35
1.35
1.36
0.01

1640
W682K
682
0.98
1.08
1.1
1.05
0.05

1641
W682I
682
0.83
1.06
0.92
0.94
0.09

1642
W682N
682
0.91
0.82
1.02
0.92
0.08

1643
W682T
682
0.88
0.88
0.97
0.91
0.04

1644
W682Q
682
0.86
0.82
1
0.89
0.08

1645
W682E
682
0.87
0.85
0.83
0.85
0.02

1646
W682A
682
0.74
0.82
0.94
0.83
0.08

1647
W682V
682
0.7
0.86
0.77
0.78
0.06

1648
W682M
682
0.7
0.72
0.67
0.7
0.02

1649
W682G
682
0.58
0.58
0.55
0.57
0.01

1650
T683F
683
1.44
1.61
1.34
1.46
0.11

1651
T683Y
683
0.59
0.73
0.69
0.67
0.06

1652
I685Y
685
1.22
0.89
1.2
1.11
0.15

1653
S686R
686
0.57
0.99
−0.22
0.45
0.5

1654
S686T
686
0.56
0.46
0.24
0.42
0.14

1655
E687N
687
0.54
0.58
0.39
0.51
0.08

1656
K688D
688
2.15
2.25
0.24
1.55
0.92

1657
K688V
688
0.42
0.44
0.47
0.44
0.02

1658
D691R
691
0.36
1.29
0.23
0.62
0.47

1659
L692M
692
0.5
0.61
0.62
0.58
0.05

1660
L693Q
693
1.87
1.34
1.83
1.68
0.24

1661
L693W
693
1.48
1.32
1.95
1.58
0.27

1662
L693G
693
0.98
1.55
1.45
1.33
0.25

1663
L693E
693
0.46
1
0.39
0.61
0.27

1664
L693M
693
0.48
0.53
0.54
0.52
0.03

1665
L693A
693
0.94
0.4
0.04
0.46
0.37

1666
L693D
693
0.54
0.53
0.27
0.45
0.13

1667
L693T
693
0.34
0.32
0.68
0.45
0.16

1668
Q694G
694
0.83
1.35
1.37
1.18
0.25

1669
Q694W
694
1.52
0.83
1.15
1.17
0.28

1670
Q694C
694
1.15
0.65
1.08
0.96
0.22

1671
Q694R
694
0.91
1.08
0.39
0.79
0.29

1672
Q694S
694
0.14
2.18
−0.02
0.77
1

1673
Q694Y
694
0.29
0.79
0.64
0.57
0.21

1674
Q694K
694
0.46
0.47
0.68
0.53
0.1

1675
Q694I
694
0.29
0.65
0.39
0.44
0.15

1676
E695S
695
2.74
2.8
1.37
2.3
0.66

1677
G697I
697
1.32
1.7
0.9
1.3
0.33

1678
G697N
697
0.98
0.62
0.67
0.76
0.16

1679
G697P
697
0.46
0.47
0.61
0.51
0.07

1680
Q698F
698
0.14
1.26
1
0.8
0.48

1681
Q698A
698
0.14
0.81
0.39
0.45
0.28

1682
L699Y
699
0.43
1.29
0.68
0.8
0.36

1683
L699F
699
0.71
0.73
0.79
0.75
0.03

1684
L699C
699
0.36
0.25
1.31
0.64
0.47

1685
Y700V
700
1.28
1.46
1.08
1.27
0.15

1686
Y700A
700
0.94
1.21
1.2
1.12
0.12

1687
Y700G
700
0.7
1.26
1.12
1.03
0.24

1688
Y700I
700
0.95
0.65
0.21
0.6
0.3

1689
Y700T
700
0.54
0.25
0.68
0.49
0.18

1690
L701P
701
1.61
1.18
1.15
1.31
0.21

1691
L701W
701
0.94
0.87
1.27
1.02
0.17

1692
F702G
702
2.82
2.75
2.85
2.81
0.04

1693
F702N
702
2.71
2.69
2.89
2.77
0.09

1694
F702A
702
2.49
2.54
2.71
2.58
0.1

1695
F702Q
702
2.31
2.03
2.4
2.25
0.16

1696
F702H
702
1.96
2
1.85
1.94
0.06

1697
F702T
702
1.72
1.73
1.7
1.72
0.01

1698
F702M
702
1.52
1.73
1.7
1.65
0.09

1699
F702S
702
1.27
1.35
1.35
1.32
0.04

1700
F702V
702
0.95
0.94
1.03
0.97
0.04

1701
F702C
702
0.55
0.79
0.7
0.68
0.1

1702
F702L
702
0.46
0.52
0.51
0.5
0.03

1703
Q703C
703
1.24
1.63
2.06
1.64
0.34

1704
K711P
711
1.73
1.48
1.74
1.65
0.12

1705
K711L
711
0.31
0.46
0.54
0.44
0.1

1706
K712W
712
2.68
2.64
2.57
2.63
0.04

1707
K712F
712
2.35
2.71
2.74
2.6
0.18

1708
K712Y
712
2.42
2.5
2.39
2.44
0.04

1709
K712G
712
2.06
2.27
1.84
2.05
0.17

1710
K712H
712
1.88
2.19
2.08
2.05
0.13

1711
K712S
712
1.11
0.95
1.2
1.09
0.1

1712
K712R
712
0.85
1.37
0.88
1.03
0.24

1713
K712C
712
0.83
0.24
1.26
0.78
0.42

1714
K712A
712
0.86
0.24
0.98
0.69
0.32

1715
K712Q
712
0.13
0.93
0.58
0.55
0.33

1716
S713R
713
1.23
0.94
−0.02
0.72
0.54

1717
T714Y
714
1.74
1.55
1.77
1.69
0.1

1718
T714W
714
0.65
0.65
0.75
0.68
0.05

1719
T714N
714
0.72
0.89
0.38
0.66
0.21

1720
N716R
716
1.71
1.88
1.96
1.85
0.11

1721
N716M
716
1.32
1.49
1.47
1.43
0.08

1722
N716I
716
1.18
1.25
1.29
1.24
0.04

1723
N716V
716
1.22
1.16
1.22
1.2
0.03

1724
N716T
716
1.04
1.22
1.12
1.12
0.07

1725
N716C
716
0.69
1.03
0.49
0.73
0.23

1726
N716Q
716
0.55
0.81
0.7
0.69
0.11

1727
N716A
716
0.53
0.49
0.45
0.49
0.04

1728
N716K
716
0.46
0.53
0.46
0.48
0.03

1729
D717P
717
2.05
2.25
2.06
2.12
0.09

1730
D717A
717
0.68
0.61
0.65
0.65
0.03

1731
L719K
719
0.42
0.58
0.63
0.54
0.09

1732
K725Y
725
0.81
0.68
0.46
0.65
0.14

1733
L727H
727
0.75
0.66
0.43
0.61
0.13

1734
L727T
727
0.14
0.64
0.89
0.56
0.31

1735
L727I
727
0.36
−0.05
0.99
0.43
0.43

1736
F728T
728
1.39
0.93
−0.02
0.76
0.59

1737
S729H
729
0.83
0.46
0.38
0.56
0.2

1738
S729K
729
1.06
0.23
−0.02
0.42
0.46

1739
E730L
730
0.92
3.32
2.58
2.28
1

1740
E730M
730
1.75
1.33
−0.02
1.02
0.76

1741
E730I
730
0.83
1.33
0.67
0.95
0.28

1742
E730G
730
1.24
0.64
0.54
0.8
0.31

1743
E730V
730
0.61
0.23
1.01
0.62
0.32

1744
E731P
731
1.47
4.46
3.64
3.19
1.26

1745
E731A
731
0.65
1.77
−0.43
0.67
0.9

1746
E731L
731
0.65
0.23
0.83
0.57
0.25

1747
N732T
732
1.36
4.47
3.65
3.16
1.31

1748
N732D
732
1.24
1.22
0.49
0.98
0.35

1749
L733I
733
0.55
0.67
0.57
0.59
0.05

1750
L733K
733
2.09
−0.08
−0.59
0.47
1.16

1751
D735R
735
0.41
0.46
0.7
0.52
0.13

1752
D735K
735
0.83
0.32
0.31
0.49
0.25

1753
V737Q
737
0.82
1.14
0.37
0.77
0.32

1754
L738E
738
1.12
1.05
1.01
1.06
0.05

1755
N741G
741
1.15
1.17
1.44
1.26
0.13

1756
N741C
741
0.48
0.47
0.96
0.64
0.23

1757
E743N
743
0.72
0.89
0.91
0.84
0.09

1758
A744N
744
3.28
3.38
3.54
3.4
0.11

1759
A744P
744
2.39
2.69
2.65
2.58
0.13

1760
A744T
744
2.4
2.61
2.67
2.56
0.11

1761
A744C
744
1.68
1.77
1.8
1.75
0.05

1762
A744S
744
1.51
1.79
1.78
1.69
0.13

1763
E745P
745
0.71
1.04
0.67
0.8
0.17

1764
F748G
748
2.41
2.55
2.55
2.5
0.07

1765
F748A
748
2.38
2.37
2.59
2.44
0.1

1766
F748S
748
2.38
2.48
2.44
2.43
0.04

1767
F748H
748
2.2
2.28
2.47
2.32
0.11

1768
F748W
748
1.84
1.93
1.89
1.89
0.04

1769
F748T
748
1.75
1.74
1.95
1.81
0.1

1770
F748R
748
1.23
1.46
1.69
1.46
0.19

1771
F748V
748
1.24
1.25
1.2
1.23
0.02

1772
F748E
748
0.84
1.2
1.33
1.12
0.2

1773
F748M
748
0.73
1.28
1.36
1.12
0.28

1774
F748N
748
1.01
1.26
0.91
1.06
0.15

1775
F748Q
748
1.07
0.89
1.22
1.06
0.14

1776
F748C
748
1.02
0.78
1.06
0.95
0.12

1777
F748D
748
1.15
1
0.6
0.92
0.23

1778
F748I
748
0.24
0.63
0.41
0.43
0.16

1779
K750P
750
3.95
4.11
4.24
4.1
0.12

1780
K750G
750
3.13
3.25
3.33
3.24
0.08

1781
K750A
750
1.95
1.96
2.04
1.98
0.04

1782
K750E
750
1.74
1.89
1.8
1.81
0.06

1783
K750W
750
1.81
1.71
1.53
1.69
0.12

1784
K750S
750
1.59
1.63
1.72
1.65
0.06

1785
K750F
750
1.56
1.58
1.6
1.58
0.02

1786
K750Y
750
1.57
1.45
1.55
1.53
0.05

1787
K750H
750
1.29
1.37
1.41
1.36
0.05

1788
K750N
750
0.9
1.04
1
0.98
0.06

1789
K750L
750
0.76
0.55
0.68
0.66
0.09

1790
K750T
750
0.67
0.62
0.61
0.63
0.03

1791
S751A
751
3.59
3.78
3.93
3.77
0.14

1792
S751G
751
3.08
3.43
3.39
3.3
0.16

1793
S751Q
751
3.01
3.09
3.16
3.09
0.06

1794
S751L
751
2.97
3.09
3.19
3.08
0.09

1795
S751H
751
2.59
2.88
3.05
2.84
0.19

1796
S751C
751
2.51
2.83
2.79
2.71
0.15

1797
S751P
751
2.62
2.76
2.75
2.71
0.06

1798
S751M
751
2.35
2.54
2.57
2.48
0.1

1799
S751N
751
2.48
2.49
2.39
2.45
0.04

1800
S751E
751
2.32
2.4
2.38
2.37
0.03

1801
S751R
751
1.89
1.92
2
1.94
0.05

1802
S751F
751
1.44
1.37
1.59
1.47
0.09

1803
S751V
751
1.39
1.4
1.36
1.39
0.02

1804
S751T
751
1.23
1.28
1.29
1.27
0.03

1805
S751D
751
0.31
0.78
0.75
0.62
0.21

1806
S752Y
752
2.59
2.71
2.67
2.66
0.05

1807
S752F
752
2.27
2.46
2.5
2.41
0.1

1808
S752H
752
2.27
2.28
2.13
2.23
0.07

1809
S752I
752
1.9
2.14
2.01
2.02
0.1

1810
S752C
752
1.57
1.7
1.68
1.65
0.06

1811
S752V
752
1.39
1.56
1.46
1.47
0.07

1812
S752M
752
1.17
1.45
1.29
1.3
0.11

1813
S752G
752
1.27
1.22
1.16
1.22
0.04

1814
S752W
752
1.17
1.16
1.26
1.2
0.05

1815
S752Q
752
1.22
1.18
1.1
1.17
0.05

1816
S752L
752
0.41
0.48
0.49
0.46
0.04

1817
I753E
753
0.36
0.45
0.45
0.42
0.04

1818
K754G
754
3.52
3.77
3.8
3.69
0.13

1819
K754H
754
3.55
3.6
3.84
3.66
0.13

1820
K754W
754
3.22
3.06
3.2
3.16
0.07

1821
K754S
754
2.91
3.1
2.99
3
0.08

1822
K754V
754
2.66
2.78
2.85
2.76
0.08

1823
K754Y
754
2.63
2.79
2.85
2.76
0.09

1824
K754I
754
2.66
2.73
2.83
2.74
0.07

1825
K754F
754
2.67
2.65
2.72
2.68
0.03

1826
K754D
754
2.22
2.57
2.59
2.46
0.17

1827
K754C
754
2.06
2.73
2.33
2.37
0.27

1828
K754A
754
2.17
2.4
2.36
2.31
0.1

1829
K754L
754
2.09
2.28
2.35
2.24
0.11

1830
K754P
754
1.84
1.77
1.72
1.78
0.05

1831
K754T
754
1.42
1.56
1.55
1.51
0.07

1832
K754N
754
0.78
0.84
0.86
0.83
0.04

1833
K754Q
754
0.68
0.79
0.77
0.75
0.05

1834
K754E
754
0.65
0.68
0.73
0.68
0.03

1835
P756M
756
2.25
1.46
1.05
1.59
0.5

1836
P756I
756
1.62
1.46
1.65
1.58
0.08

1837
P756K
756
1.37
1.71
1.45
1.51
0.15

1838
P756H
756
1.03
1.31
1.61
1.32
0.24

1839
P756N
756
0.11
1.6
2.03
1.25
0.82

1840
P756L
756
0.97
1.4
0.84
1.07
0.24

1841
P756S
756
0.9
0.68
1.11
0.9
0.18

1842
P756T
756
0.44
0.82
0.91
0.72
0.21

1843
I757W
757
3.29
3.3
4.2
3.6
0.43

1844
I757P
757
2.89
3.01
2.85
2.91
0.07

1845
I757K
757
2.34
2.35
2.55
2.42
0.1

1846
I757F
757
1.95
2.16
2.3
2.14
0.14

1847
I757A
757
1.96
2.1
1.85
1.97
0.1

1848
I757R
757
1.45
1.91
1.45
1.6
0.22

1849
I757Y
757
1.72
1.75
1.25
1.57
0.23

1850
I757L
757
1.1
1.3
1.17
1.19
0.09

1851
I757S
757
0.81
1.06
0.87
0.91
0.11

1852
I757Q
757
0.85
0.1
1.11
0.69
0.43

1853
I757C
757
0.52
−0.08
1.2
0.55
0.53

1854
I757G
757
0.35
0.58
0.5
0.48
0.09

1855
I757M
757
0.36
0.6
0.45
0.47
0.1

1856
I758A
758
1.77
1.91
1.77
1.82
0.07

1857
I758E
758
1.58
1.84
1.86
1.76
0.13

1858
I758G
758
1.39
1.66
1.48
1.51
0.11

1859
I758Q
758
1.66
1.29
1.39
1.45
0.16

1860
I758S
758
1.33
1.46
1.39
1.39
0.05

1861
I758T
758
0.97
1.07
1.44
1.16
0.2

1862
I758C
758
1.04
1.01
1.15
1.07
0.06

1863
I758V
758
0.95
1.06
1.03
1.01
0.05

1864
I758K
758
0.85
0.82
0.69
0.79
0.07

1865
G762K
762
0.46
0.05
0.89
0.47
0.34

1866
I764Q
764
0.7
0.75
0.49
0.65
0.11

1867
V766Y
766
0.51
0.89
−0.06
0.45
0.39

1868
N767R
767
0.25
0.56
0.46
0.43
0.13

1869
R768K
768
1.4
0.89
1.48
1.26
0.26

1870
T769L
769
0.46
0.35
0.57
0.46
0.09

1871
E771W
771
1.53
1.44
1.65
1.54
0.09

1872
E771A
771
0.88
0.86
0.95
0.9
0.04

1873
E771R
771
0.77
0.75
0.58
0.7
0.09

1874
E771C
771
0.76
0.6
0.7
0.69
0.06

1875
E771Y
771
0.56
0.73
0.69
0.66
0.07

1876
E771I
771
0.7
0.68
0.36
0.58
0.16

1877
E771T
771
0.62
0.59
0.53
0.58
0.04

1878
E771F
771
0.52
0.75
0.43
0.57
0.14

1879
E771K
771
0.48
0.53
0.66
0.56
0.08

1880
E771M
771
0.5
0.43
0.69
0.54
0.11

1881
E771S
771
0.48
0.56
0.51
0.52
0.03

1882
E771H
771
0.47
0.38
0.69
0.51
0.13

1883
E771Q
771
0.37
0.64
0.49
0.5
0.11

1884
E771V
771
0.26
0.58
0.41
0.42
0.13

1885
A772W
772
0.79
0.19
0.89
0.62
0.31

1886
E773I
773
0.88
0.36
0.58
0.6
0.22

1887
E773P
773
0.68
0.48
0.25
0.47
0.17

1888
E774W
774
1.49
1.89
1.73
1.7
0.17

1889
E774M
774
1.75
1.58
1.27
1.53
0.2

1890
E774S
774
0.96
0.73
0.66
0.78
0.13

1891
E774V
774
0.51
0.88
0.93
0.77
0.19

1892
E774T
774
0.54
0.88
0.72
0.71
0.14

1893
E774K
774
0.54
0.74
0.74
0.67
0.09

1894
E774L
774
0.87
0.43
0.65
0.65
0.18

1895
E774I
774
0.8
0.78
0.12
0.56
0.32

1896
E774R
774
0.71
0.24
0.6
0.52
0.2

1897
E774Q
774
0.84
0.34
0.28
0.49
0.25

1898
E774H
774
0.8
0.39
0.22
0.47
0.24

1899
K775C
775
0.7
0.51
0.34
0.51
0.15

1900
Q777R
777
0.84
0.57
0.54
0.65
0.13

1901
Q777V
777
0.46
0.72
0.67
0.62
0.11

1902
F778Q
778
0.87
0.47
1.08
0.81
0.25

1903
F778K
778
0.92
0.68
0.48
0.69
0.18

1904
F778R
778
0.46
0.61
0.43
0.5
0.08

1905
F778G
778
0.57
0.28
0.4
0.42
0.12

1906
I783Q
783
0.61
1.03
0.91
0.85
0.18

1907
V784F
784
1.03
1.39
1.33
1.25
0.16

1908
V784L
784
1.06
0.92
1.09
1.03
0.07

1909
V784Y
784
0.81
1.23
0.9
0.98
0.18

1910
V784R
784
0.73
0.93
0.99
0.88
0.11

1911
V784W
784
0.47
0.74
1.04
0.75
0.23

1912
V784H
784
−0.58
0.89
1.33
0.55
0.82

1913
V784I
784
0.6
0.42
0.43
0.48
0.08

1914
N787A
787
3.01
3.25
3.28
3.18
0.12

1915
N787W
787
2.32
2.6
2.15
2.36
0.19

1916
N787G
787
2.43
2.15
1.9
2.16
0.22

1917
N787Q
787
2.07
2.4
1.56
2.01
0.35

1918
N787P
787
1.92
1.98
1.98
1.96
0.03

1919
N787R
787
1.92
1.65
1.92
1.83
0.13

1920
N787S
787
1.73
1.74
1.95
1.81
0.1

1921
N787C
787
2.07
0.9
1.56
1.51
0.48

1922
N787M
787
1.28
1.36
1.53
1.39
0.1

1923
N787H
787
0.68
1.46
1.05
1.06
0.32

1924
N787F
787
0.76
0.84
0.97
0.86
0.09

1925
I788Y
788
0.53
0.79
1.15
0.82
0.25

1926
I788F
788
0.57
0.49
0.7
0.59
0.09

1927
P789C
789
−0.97
1.51
0.8
0.44
1.04

1928
E790I
790
1.92
2.22
2.26
2.13
0.15

1929
E790P
790
1.59
1.82
2.22
1.88
0.26

1930
E790F
790
1.74
1.59
1.75
1.69
0.07

1931
E790V
790
1.82
1.64
1.45
1.64
0.15

1932
E790R
790
1.25
1.64
1.79
1.56
0.23

1933
E790K
790
1.47
1.59
1.51
1.52
0.05

1934
E790A
790
1.43
1.38
1.75
1.52
0.16

1935
E790T
790
1.71
0.9
1.82
1.47
0.41

1936
E790N
790
1.71
1.48
1.05
1.41
0.27

1937
E790S
790
1.64
1.11
1.05
1.27
0.26

1938
E790Q
790
0.96
1.57
1.08
1.2
0.26

1939
E790W
790
1.08
1.43
0.83
1.11
0.25

1940
E790M
790
1.38
1.02
0.92
1.11
0.2

1941
E790C
790
1.51
1.82
−0.05
1.09
0.82

1942
E790G
790
1.07
0.97
0.9
0.98
0.07

1943
E790Y
790
1.13
0.79
0.68
0.87
0.19

1944
E790H
790
1.08
0.74
0.22
0.68
0.36

1945
N791G
791
2.07
0.2
0.65
0.97
0.8

1946
N791E
791
0.29
0.85
0.6
0.58
0.23

1947
N791F
791
0.57
0.65
0.4
0.54
0.1

1948
Y793V
793
1.31
0.7
1.27
1.09
0.28

1949
Y793R
793
0.7
0.95
0.59
0.75
0.15

1950
Y793W
793
0.31
0.79
0.83
0.64
0.24

1951
Y793Q
793
0.82
0.49
0.36
0.56
0.19

1952
Q794P
794
1
1.43
0.14
0.86
0.54

1953
Q794L
794
0.5
0.94
0.53
0.66
0.2

1954
Y797W
797
0.64
1.05
−0.22
0.49
0.53

1955
K798L
798
1.17
0.46
0.78
0.8
0.29

1956
F800W
800
0.82
1.31
0.94
1.02
0.21

1957
N801A
801
0.82
0.9
1.35
1.02
0.23

1958
N801S
801
0.13
−0.48
1.66
0.44
0.9

1959
K803A
803
1.23
1.6
0.65
1.16
0.39

1960
S804F
804
1.23
0.91
0.88
1
0.16

1961
S804I
804
0.42
1.51
−0.04
0.63
0.65

1962
K806C
806
0.2
0.75
0.54
0.5
0.23

1963
S809G
809
0.59
0.27
0.62
0.49
0.16

1964
A813H
813
0.78
1.11
0.24
0.71
0.36

1965
E823P
823
1.14
0.88
0.84
0.96
0.13

1966
E823A
823
0.67
0.5
0.69
0.62
0.09

1967
E823S
823
0.19
0.88
0.19
0.42
0.33

1968
A825H
825
1.51
1.71
1.82
1.68
0.13

1969
A825R
825
1.11
1.34
1.4
1.28
0.12

1970
A825K
825
0.38
0.53
0.45
0.45
0.06

1971
T826F
826
3.15
3.35
3.35
3.28
0.1

1972
T826Y
826
3.03
3.3
3.28
3.2
0.12

1973
T826W
826
2.55
2.75
2.62
2.64
0.08

1974
T826H
826
1.94
2
2.23
2.06
0.12

1975
T826C
826
1.73
1.79
1.96
1.83
0.1

1976
T826I
826
1.56
1.77
1.89
1.74
0.13

1977
T826M
826
1.45
1.67
1.7
1.61
0.11

1978
T826L
826
1.1
1.3
1.4
1.27
0.12

1979
T826K
826
1.15
1.29
1.27
1.23
0.06

1980
T826R
826
1.07
1.13
1.24
1.15
0.07

1981
T826Q
826
0.89
0.72
0.8
0.8
0.07

1982
T826N
826
0.29
0.48
0.5
0.42
0.09

1983
N827D
827
0.95
0.69
0.72
0.79
0.12

1984
N827E
827
0.58
0.84
0.77
0.73
0.11

1985
V829T
829
1.87
1.96
1.9
1.91
0.04

1986
D831L
831
2.04
2.27
2.27
2.19
0.11

1987
D831M
831
0.83
0.75
0.97
0.85
0.09

1988
D831Q
831
1.12
0.84
0.39
0.78
0.3

1989
D831R
831
0.83
1.17
−0.13
0.63
0.55

1990
D831N
831
0.83
0.7
0.1
0.54
0.32

1991
Y832R
832
3.17
3.35
3.45
3.32
0.12

1992
Y832K
832
2.7
2.97
3.04
2.9
0.15

1993
Y832G
832
2.08
2.22
2.29
2.2
0.09

1994
Y832Q
832
1.46
1.52
1.59
1.53
0.05

1995
Y832A
832
1.01
1.2
1.15
1.12
0.08

1996
Y832N
832
0.96
1.12
1.06
1.04
0.07

1997
Y832M
832
0.68
0.81
0.89
0.79
0.09

1998
Y832E
832
0.5
0.63
0.68
0.6
0.08

1999
Y836K
836
3.67
3.99
4.02
3.89
0.16

2000
Y836Q
836
3.61
3.78
3.87
3.75
0.1

2001
Y836R
836
3.11
3.28
3.31
3.23
0.09

2002
Y836I
836
3.21
2.99
3.08
3.09
0.09

2003
Y836D
836
2.87
2.91
3.25
3.01
0.17

2004
Y836V
836
2.83
3.07
3.08
3
0.12

2005
Y836T
836
2.81
3.05
2.89
2.92
0.1

2006
Y836N
836
2.71
2.95
3.04
2.9
0.14

2007
Y836E
836
2.67
3.07
2.76
2.84
0.17

2008
Y836H
836
2.89
2.92
2.68
2.83
0.1

2009
Y836L
836
2.48
2.69
2.81
2.66
0.14

2010
Y836M
836
2.61
2.72
2.65
2.66
0.05

2011
Y836A
836
2.7
2.43
2.57
2.57
0.11

2012
Y836S
836
2.09
2.37
2.37
2.28
0.13

2013
Y836G
836
1.93
1.69
1.94
1.85
0.12

2014
Y836C
836
0.84
0.93
0.83
0.86
0.04

2015
D837P
837
2.8
2.8
2.83
2.81
0.02

2016
Y839L
839
2.9
3.07
3.09
3.02
0.08

2017
Y839H
839
2.72
2.39
2.54
2.55
0.14

2018
Y839I
839
2.47
2.31
2.69
2.49
0.15

2019
Y839T
839
2.38
2.25
2.51
2.38
0.11

2020
Y839Q
839
2.34
2.25
2.46
2.35
0.09

2021
Y839V
839
2.1
2.35
2.31
2.25
0.11

2022
Y839M
839
2.11
2.12
2.31
2.18
0.09

2023
Y839K
839
2.16
2.04
2.23
2.15
0.08

2024
Y839C
839
1.75
2.16
2.02
1.98
0.17

2025
Y839R
839
2.1
1.79
1.93
1.94
0.13

2026
Y839N
839
1.81
1.83
2.07
1.91
0.12

2027
Y839A
839
0.73
0.93
1.42
1.03
0.29

2028
Y839S
839
0.58
0.82
0.72
0.71
0.1

2029
L841I
841
1.5
1.85
1.53
1.63
0.15

2030
L841V
841
0.91
0.89
1
0.94
0.05

2031
L841C
84
0.78
0.87
1.09
0.91
0.13

2032
L841G
841
0.5
1
0.75
0.75
0.2

2033
L841Y
841
0.47
0.53
0.62
0.54
0.06

2034
M843F
843
1.65
1.94
2.27
1.95
0.25

2035
M843Q
843
1.56
1.34
1.64
1.51
0.13

2036
P844C
844
0.14
0.93
1.37
0.82
0.51

2037
I845Q
845
1.75
0.24
−0.01
0.66
0.78

2038
T846G
846
1.53
1.34
1.69
1.52
0.15

2039
T846Q
846
1.4
0.93
1.6
1.31
0.28

2040
T846Y
846
0.84
0.71
0.32
0.62
0.22

2041
T846L
846
0.59
0.63
0.44
0.55
0.08

2042
T846W
846
0.5
0.69
0.12
0.44
0.24

2043
N848D
848
1.06
0.93
0.91
0.97
0.07

2044
F849R
849
0.41
0.71
1.02
0.71
0.25

2045
A851T
851
1.06
0.5
0.68
0.75
0.23

2046
A851N
851
0.14
0.93
0.68
0.59
0.33

2047
I857A
857
0.66
0.24
0.5
0.46
0.17

2048
I857N
857
0.14
0.75
0.5
0.46
0.25

2049
D859P
859
0.43
1.22
1.38
1.01
0.41

2050
R860E
860
0.15
0.46
1.44
0.68
0.55

2051
R860K
860
0.84
0.73
0.21
0.59
0.27

2052
I861A
861
0.44
0.49
0.56
0.5
0.05

2053
L862Y
862
1.37
1.72
1.08
1.39
0.26

2054
L862D
862
0.7
0.46
0.9
0.69
0.18

2055
L862H
862
0.8
0.69
0.12
0.53
0.3

2056
A866D
866
1.44
0.93
1.37
1.25
0.23

2057
A866Y
866
0.6
0.39
0.63
0.54
0.11

2058
K869C
869
0.54
0.79
0.79
0.71
0.12

2059
H872S
872
1.05
0.3
0.2
0.51
0.38

2060
H872M
872
0.91
0.52
−0.02
0.47
0.38

2061
I876L
876
0.71
0.67
0.63
0.67
0.03

2062
V886S
886
0.88
0.97
0.89
0.91
0.04

2063
V886A
886
0.57
0.58
0.56
0.57
0.01

2064
V886G
886
0.56
0.63
0.48
0.56
0.06

2065
T891I
891
0.31
0.82
0.4
0.51
0.22

2066
C892T
892
1.35
1.22
1.45
1.34
0.09

2067
C892I
892
1.06
0.93
0.54
0.85
0.22

2068
C892D
892
0.84
2.19
−0.71
0.77
1.18

2069
C892H
892
0.15
1.09
0.83
0.69
0.4

2070
G893K
893
0.99
0.24
0.27
0.5
0.35

2071
N894V
894
0.42
1.06
0.91
0.8
0.28

2072
E897Y
897
0.53
0.48
1
0.67
0.23

2073
Q898G
898
0.84
0.93
−0.02
0.58
0.43

2074
F901M
901
2.43
2.67
2.59
2.56
0.1

2075
F901Q
901
2.09
2.12
1.9
2.04
0.1

2076
F901L
901
0.61
0.71
0.75
0.69
0.06

2077
F901G
901
0.48
0.82
0.62
0.64
0.14

2078
I903V
903
0.36
0.43
0.47
0.42
0.05

2079
Y907I
907
3.11
3.15
3.13
3.13
0.02

2080
Y907T
907
1.74
1.42
1.12
1.43
0.25

2081
Y907M
907
0.84
1.1
1.8
1.25
0.41

2082
Y907V
907
0.9
0.8
1.15
0.95
0.15

2083
Y909F
909
1.11
1.01
1.31
1.14
0.12

2084
Y909W
909
1.02
1
1.13
1.05
0.06

2085
Q910A
910
0.83
0.46
1.35
0.88
0.37

2086
Q910N
910
1.24
0.57
0.56
0.79
0.32

2087
Q910Y
910
0.93
0.77
0.06
0.59
0.38

2088
Q910S
910
0.65
0.84
0.12
0.54
0.3

2089
I911Q
911
1.29
1.66
1.54
1.5
0.15

2090
I911E
911
1.41
1.31
1.59
1.44
0.12

2091
I911A
911
1.24
1.25
1.32
1.27
0.03

2092
I911H
911
0.95
0.95
0.83
0.91
0.06

2093
I911D
911
0.53
0.9
1
0.81
0.2

2094
I911Y
911
0.7
0.89
0.59
0.73
0.12

2095
I911G
911
0.65
0.49
0.54
0.56
0.07

2096
I911W
911
0.5
0.4
0.66
0.52
0.11

2097
K912H
912
1.08
0.93
0.69
0.9
0.16

2098
K912Y
912
0.79
0.95
0.51
0.75
0.18

2099
K912C
912
1.03
−0.73
1.04
0.45
0.83

2100
K914A
914
1.64
1.3
1.54
1.49
0.14

2101
K914S
914
1.11
0.97
1.01
1.03
0.06

2102
K914Q
914
0.75
0.88
0.97
0.86
0.09

2103
K914F
914
0.89
0.73
0.87
0.83
0.07

2104
K914T
914
0.74
0.65
0.63
0.67
0.05

2105
K914M
914
0.7
0.54
0.68
0.64
0.07

2106
K914C
914
0.68
0.39
0.64
0.57
0.13

2107
Q916Y
916
1.04
1.29
1.02
1.12
0.12

2108
Q916M
916
0.77
1.08
1.16
1
0.17

2109
Q916D
916
1.06
0.5
1.13
0.89
0.28

2110
Q916V
916
0.82
0.62
1.05
0.83
0.18

2111
Q916L
916
0.62
0.79
0.7
0.7
0.07

2112
Q916A
916
0.65
0.85
0.6
0.7
0.11

2113
Q916N
916
0.87
0.15
0.73
0.59
0.31

2114
Q916W
916
0.45
0.76
0.51
0.58
0.14

2115
G918E
918
1.98
1.96
2.03
1.99
0.03

2116
G918M
918
1.82
1.68
1.66
1.72
0.07

2117
G918Q
918
1.29
1.44
1.38
1.37
0.06

2118
G918L
918
1.08
1.32
1.45
1.28
0.15

2119
G918I
918
1.16
1.37
1.3
1.28
0.09

2120
G918T
918
0.76
0.89
0.89
0.85
0.06

2121
G918Y
918
0.58
0.91
0.62
0.71
0.14

2122
G918F
918
0.68
0.63
0.61
0.64
0.03

2123
G918H
918
0.4
0.92
0.6
0.64
0.21

2124
G918K
918
0.62
0.52
0.72
0.62
0.08

2125
G918D
918
0.48
0.42
0.48
0.46
0.03

2126
A919I
919
1.99
2.11
1.95
2.02
0.07

2127
A919E
919
1.36
0.59
1.19
1.05
0.33

2128
A919F
919
0.83
0.89
0.92
0.88
0.04

2129
A919L
919
0.84
0.78
0.83
0.82
0.03

2130
A919Y
919
0.28
1.14
0.76
0.73
0.35

2131
A919M
919
0.51
0.57
0.33
0.47
0.1

2132
Q921A
921
0.83
0.55
0.47
0.62
0.15

2133
Q921T
921
0.74
0.23
0.65
0.54
0.22

2134
I922A
922
0.71
0.55
0.76
0.67
0.09

2135
I922R
922
0.55
0.73
0.58
0.62
0.08

2136
I922Y
922
0.46
0.63
0.57
0.55
0.07

2137
I922W
922
0.38
0.68
0.49
0.51
0.13

2138
I922C
922
0.34
0.64
0.37
0.45
0.14

2139
I922H
922
0.39
0.47
0.44
0.43
0.03

2140
K925C
925
0.82
0.39
0.65
0.62
0.18

2141
E926S
926
1.06
1.15
1.18
1.13
0.05

2142
E926T
926
0.97
0.99
0.98
0.98
0.01

2143
E926N
926
0.44
0.48
0.36
0.43
0.05

2144
K928W
928
0.78
0.7
0.67
0.72
0.05

2145
E929C
929
1.05
0.92
1.21
1.06
0.12

2146
E929G
929
0.74
0.67
0.8
0.74
0.05

2147
E929S
929
0.42
0.52
0.61
0.52
0.07

2148
E929P
929
0.41
0.5
0.62
0.51
0.09

2149
E929N
929
0.57
0.31
0.61
0.5
0.13

2150
G931Q
931
0.54
0.64
0.98
0.72
0.19

2151
G931H
931
0.25
0.62
0.43
0.43
0.15

2152
K934S
934
0.64
0.38
0.6
0.54
0.12

2153
E935R
935
2.72
2.95
3.05
2.91
0.14

2154
E935M
935
2.9
2.77
2.97
2.88
0.08

2155
E935N
935
2.75
2.91
2.8
2.82
0.07

2156
E935A
935
2.43
2.62
2.7
2.58
0.11

2157
E935Q
935
2.21
2.5
2.62
2.44
0.17

2158
E935K
935
2.25
2.53
2.53
2.43
0.13

2159
E935L
935
1.91
2.07
2.1
2.03
0.08

2160
E935S
935
1.75
2.08
1.96
1.93
0.14

2161
E935Y
935
1.93
1.85
1.75
1.84
0.07

2162
E935H
935
1.73
2.03
1.76
1.84
0.14

2163
E935C
935
1.39
1.55
1.56
1.5
0.08

2164
E935G
935
1.1
1.58
1.35
1.34
0.2

2165
E935F
935
1.18
1.5
1.34
1.34
0.13

2166
E935T
935
1.2
1.36
1.46
1.34
0.11

2167
E935W
935
0.78
1.29
1.5
1.19
0.3

2168
I936T
936
2.76
2.88
2.96
2.86
0.08

2169
I936C
936
2.11
1.85
1.91
1.96
0.11

2170
I936A
936
1.94
1.92
1.99
1.95
0.03

2171
I936R
936
1.72
1.75
1.81
1.76
0.04

2172
I936K
936
1.57
1.63
1.66
1.62
0.03

2173
I936S
936
1.38
1.37
1.44
1.4
0.03

2174
I936Q
936
1.16
1.48
1.29
1.31
0.13

2175
I936Y
936
1.17
1.18
1.25
1.2
0.03

2176
I936F
936
0.99
0.83
0.99
0.94
0.08

2177
I936L
936
0.99
0.85
0.92
0.92
0.06

2178
I936G
936
0.59
0.11
0.56
0.42
0.22

2179
L941C
941
1.77
1.64
1.64
1.68
0.06

2180
L941N
941
1.06
0.63
0.65
0.78
0.2

2181
S942A
942
1.93
2.14
1.99
2.02
0.09

2182
L943I
943
1.24
2.17
1.9
1.77
0.39

2183
V944C
944
0.63
0.58
0.6
0.6
0.02

2184
I945M
945
0.99
1.24
1.18
1.14
0.11

2185
H946W
946
1.36
1.51
1.36
1.41
0.07

2186
E947P
947
1.57
1.71
1.88
1.72
0.13

2187
E947F
947
0.49
0.47
0.74
0.56
0.13

2188
E947M
947
0.35
0.35
0.65
0.45
0.14

2189
I948L
948
1.03
0.84
0.77
0.88
0.11

2190
S949M
949
1.72
1.74
1.84
1.77
0.05

2191
S949V
949
1.54
1.69
1.58
1.6
0.06

2192
S949T
949
1.08
1.32
1.28
1.23
0.11

2193
S949Q
949
1.43
1.02
1.12
1.19
0.18

2194
S949I
949
0.9
0.66
1.02
0.86
0.15

2195
S949K
949
0.47
0.34
0.65
0.49
0.12

2196
M951G
951
−0.15
0.79
0.77
0.47
0.44

2197
V952E
952
0.79
0.38
0.73
0.63
0.18

2198
V952W
952
0.78
0.41
0.6
0.6
0.15

2199
I953A
953
0.62
0.47
0.82
0.63
0.14

2200
I953P
953
0.68
0.51
0.52
0.57
0.08

2201
K954A
954
0.7
1.33
1.71
1.25
0.42

2202
K954P
954
1.13
0.84
0.66
0.88
0.2

2203
Y955W
955
0.93
1.26
1.65
1.28
0.29

2204
Y955C
955
−0.55
1.15
1.22
0.61
0.82

2205
N956C
956
2.28
1.48
1.22
1.66
0.45

2206
N956W
956
1.46
1.04
1.47
1.32
0.2

2207
N956F
956
1.06
1.3
0.92
1.09
0.16

2208
N956Y
956
0.87
0.72
0.63
0.74
0.1

2209
A957P
957
1.02
1.17
1.14
1.11
0.06

2210
A957F
957
1.53
0.52
0.81
0.95
0.42

2211
A957T
957
0.32
0.93
0.45
0.56
0.26

2212
I958V
958
0.83
0.71
0.4
0.65
0.18

2213
I959F
959
1.49
1.01
0.95
1.15
0.24

2214
D963C
963
1.39
−0.46
0.37
0.44
0.76

2215
F968W
968
0.29
0.56
0.63
0.49
0.15

2216
K970C
970
0.43
0.68
0.26
0.46
0.17

2217
K974G
974
3.37
3.23
3.31
3.31
0.06

2218
K974P
974
2.45
2.32
2.38
2.38
0.06

2219
K974W
974
1.25
2.18
1.77
1.74
0.38

2220
K974A
974
1.12
1.29
1.62
1.34
0.21

2221
K974T
974
1.29
1.44
0.92
1.22
0.22

2222
K974Q
974
0.71
1.22
1.08
1
0.21

2223
K974Y
974
0.99
0.56
1.44
1
0.36

2224
K974C
974
1.16
0.46
1.23
0.95
0.35

2225
K974S
974
0.95
0.69
0.67
0.77
0.12

2226
K974M
974
0.5
0.99
−0.08
0.47
0.44

2227
K974E
974
0.64
−0.23
0.95
0.45
0.5

2228
V975I
975
0.66
0.6
0.69
0.65
0.04

2229
R977H
977
0.59
0.54
0.28
0.47
0.14

2230
K982Y
982
0.78
1.03
0.37
0.73
0.27

2231
K982S
982
0.56
0.69
0.75
0.67
0.08

2232
F983A
983
0.76
0.4
0.71
0.62
0.16

2233
M986G
986
0.51
0.56
0.22
0.43
0.15

2234
I988K
988
1.54
1.35
1.94
1.61
0.25

2235
I988E
988
0.85
1.63
0.68
1.06
0.41

2236
L991W
991
1.25
1.34
1.93
1.51
0.3

2237
L991C
991
1.54
0.93
1.37
1.28
0.26

2238
N992L
992
1.41
0.93
−0.02
0.78
0.59

2239
N992A
992
1.25
0.24
−0.02
0.49
0.55

2240
L994A
994
1.77
1.63
−0.01
1.13
0.81

2241
V995P
995
0.85
0.65
0.39
0.63
0.19

2242
F996T
996
0.85
0.24
0.68
0.59
0.26

2243
F996V
996
0.85
0.93
−0.01
0.59
0.43

2244
D998T
998
1.91
1.74
1.6
1.75
0.13

2245
D998W
998
0.49
0.02
0.86
0.46
0.35

2246
I999P
999
0.85
2.19
1.59
1.54
0.55

2247
I999H
999
0.16
0.65
1.08
0.63
0.38

2248
I1001W
1001
0.85
0.24
0.68
0.59
0.26

2249
T1002V
1002
1.26
0.94
−0.02
0.73
0.54

2250
T1002C
1002
1.07
−0.45
0.9
0.51
0.68

2251
T1002L
1002
0.35
0.44
0.71
0.5
0.15

2252
T1002S
1002
0.74
0.25
0.39
0.46
0.21

2253
E1003S
1003
1.62
1.09
0.97
1.23
0.29

2254
E1003M
1003
0.16
0.58
1.21
0.65
0.43

2255
N1004A
1004
2.17
1.85
−0.71
1.11
1.29

2256
N1004G
1004
0.85
0.65
1.08
0.86
0.18

2257
N1004R
1004
0.67
0.85
0.44
0.65
0.17

2258
N1004P
1004
0.63
−0.27
1.21
0.52
0.61

2259
N1004Q
1004
0.34
0.94
0.17
0.48
0.33

2260
L1007V
1007
0.5
0.72
0.48
0.57
0.11

2261
L1007C
1007
0.38
0.69
0.5
0.53
0.13

2262
L1007A
1007
0.41
0.65
0.39
0.49
0.12

2263
L1008F
1008
0.86
0.47
1.08
0.81
0.25

2264
Y1011V
1011
0.64
1.12
1.21
0.99
0.25

2265
Y1011L
1011
0.84
0.94
0.63
0.8
0.13

2266
Y1011I
1011
0.3
1.01
0.81
0.71
0.3

2267
T1014C
1014
0.54
1.15
0.77
0.82
0.25

2268
Y1015M
1015
1.1
1.28
1.29
1.22
0.09

2269
Y1015A
1015
0.9
1.19
1.14
1.08
0.13

2270
Y1015E
1015
0.99
0.82
1.06
0.96
0.1

2271
Y1015Q
1015
0.63
0.97
0.98
0.86
0.16

2272
Y1015G
1015
0.7
0.66
0.56
0.64
0.06

2273
Y1015T
1015
0.45
0.5
0.45
0.47
0.02

2274
I1016E
1016
0.96
0.94
0.92
0.94
0.02

2275
I1016N
1016
1.09
0.8
0.71
0.87
0.16

2276
I1016A
1016
0.51
0.74
0.78
0.67
0.12

2277
I1016P
1016
0.43
0.56
0.5
0.5
0.05

2278
P1017W
1017
1.42
0.66
0.79
0.95
0.33

2279
P1017G
1017
0.92
1.35
0.46
0.91
0.36

2280
P1017N
1017
1.16
1.25
0.23
0.88
0.46

2281
P1017E
1017
1.37
0.4
0.27
0.68
0.49

2282
P1017D
1017
0.3
0.7
0.86
0.62
0.24

2283
P1017Q
1017
0.91
0.35
0.44
0.56
0.25

2284
K1019A
1019
0.86
0.92
1.06
0.95
0.09

2285
K1019G
1019
0.87
0.4
0.8
0.69
0.21

2286
K1019V
1019
0.56
0.62
0.5
0.56
0.05

2287
K1019S
1019
0.45
0.51
0.62
0.53
0.07

2288
K1019C
1019
0.17
0.86
0.23
0.42
0.31

2289
K1021A
1021
1.26
1.2
1.01
1.16
0.11

2290
K1021V
1021
0.86
1.11
0.67
0.88
0.18

2291
K1021F
1021
0.42
0.78
0.43
0.54
0.17

2292
K1021W
1021
0.68
0.06
0.75
0.5
0.31

2293
V1023N
1023
1.01
0.76
0.49
0.75
0.22

2294
V1023C
1023
0.33
0.8
0.57
0.57
0.19

2295
G1024Y
1024
0.2
0.7
0.67
0.52
0.23

2296
Q1026M
1026
1.26
0.9
1.19
1.12
0.16

2297
Q1026N
1026
0.86
0.65
0.81
0.77
0.09

2298
C1027N
1027
0.58
0.64
0.65
0.62
0.03

2299
C1029N
1029
0.4
0.54
0.6
0.51
0.09

2300
Y1032Q
1032
0.84
0.84
0.73
0.8
0.05

2301
Y1032R
1032
0.86
0.72
0.7
0.76
0.07

2302
Y1032K
1032
0.83
0.6
0.82
0.75
0.11

2303
Y1032W
1032
0.64
0.14
0.67
0.49
0.24

2304
A1036H
1036
0.93
0.65
0.55
0.71
0.16

2305
A1036F
1036
0.79
0.71
0.52
0.67
0.11

2306
K1040A
1040
2.24
0.93
2.37
1.85
0.65

2307
K1040T
1040
1.01
1.91
−0.44
0.83
0.97

2308
K1040I
1040
1.26
0.23
0.66
0.72
0.42

2309
I1050D
1050
1.08
0.98
1.06
1.04
0.04

2310
I1050W
1050
0.65
0.69
0.54
0.63
0.06

2311
I1050H
1050
0.5
0.87
0.31
0.56
0.23

2312
K1052W
1052
0.76
1.25
0.94
0.98
0.2

2313
K1052C
1052
0.45
0.7
1.08
0.74
0.26

2314
K1052G
1052
0.44
0.27
0.56
0.42
0.12

2315
K1052I
1052
0.01
0.55
0.71
0.42
0.3

2316
F1053Y
1053
−0.04
1.45
−0.02
0.46
0.7

2317
K1054L
1054
1.1
1.21
1.17
1.16
0.05

2318
K1054I
1054
1.1
1.12
1
1.07
0.05

2319
K1054W
1054
0.73
1.15
0.76
0.88
0.19

2320
K1054V
1054
0.88
0.72
0.97
0.86
0.1

2321
K1054F
1054
0.94
0.52
0.73
0.73
0.17

2322
K1054M
1054
0.62
0.53
0.58
0.58
0.04

2323
D1055I
1055
1.78
1.49
1.85
1.71
0.15

2324
D1055M
1055
1.4
1.39
1.08
1.29
0.15

2325
D1055F
1055
0.93
1.63
1.21
1.26
0.29

2326
D1055T
1055
0.96
1.07
0.89
0.97
0.07

2327
D1055P
1055
0.57
1.22
0.99
0.93
0.27

2328
D1055C
1055
0.66
0.73
0.88
0.76
0.09

2329
D1055L
1055
0.3
0.86
0.98
0.71
0.3

2330
D1055V
1055
0.41
0.62
0.46
0.5
0.09

2331
D1055Q
1055
0.27
0.53
0.51
0.44
0.12

2332
L1056E
1056
0.86
0.94
0.9
0.9
0.03

2333
L1056C
1056
0.17
0.43
0.94
0.51
0.32

2334
L1056I
1056
0.24
0.5
0.78
0.5
0.22

2335
T1057Y
1057
0.57
0.65
0.67
0.63
0.04

2336
V1058W
1058
1.96
1.35
1.37
1.56
0.28

2337
V1058H
1058
0.62
0.25
0.92
0.6
0.28

2338
A1060N
1060
1.71
1.35
0.27
1.11
0.61

2339
A1060Q
1060
0.86
0.54
0.27
0.56
0.24

2340
K1061W
1061
1.96
2.04
2.28
2.1
0.14

2341
K1061F
1061
1.78
1.35
1.08
1.4
0.29

2342
K1061D
1061
0.86
0.76
1.08
0.9
0.13

2343
K1061E
1061
0.78
0.76
1.13
0.89
0.17

2344
K1061C
1061
1.09
1.17
0.39
0.88
0.35

2345
K1061S
1061
0.46
0.94
1.13
0.84
0.28

2346
K1061L
1061
0.57
0.61
0.52
0.57
0.04

2347
K1061R
1061
0.96
0.43
0.11
0.5
0.35

2348
K1061I
1061
0.54
0.69
0.18
0.47
0.21

2349
K1061Q
1061
0.86
0.54
−0.02
0.46
0.36

2350
K1061T
1061
0.51
0.86
−0.12
0.42
0.41

2351
R1062D
1062
2.47
1.35
1.08
1.64
0.6

2352
R1062A
1062
1.81
1.3
1.03
1.38
0.33

2353
R1062M
1062
1.02
1.64
0.27
0.98
0.56

2354
R1062Y
1062
1.47
0.77
0.68
0.97
0.36

2355
R1062H
1062
0.35
0.95
1.08
0.8
0.32

2356
R1062V
1062
0.87
1.48
−0.24
0.7
0.71

2357
E1063W
1063
0.31
0.51
1.04
0.62
0.31

2358
E1063F
1063
0.79
0.09
0.42
0.43
0.29

2359
E1063G
1063
0.67
−0.19
0.78
0.42
0.43

2360
I1065W
1065
0.26
0.63
0.45
0.45
0.15

2361
F1068N
1068
0.08
0.95
0.36
0.47
0.36

2362
D1069M
1069
0.4
0.66
1.38
0.81
0.41

2363
S1070E
1070
0.4
0.53
1.09
0.67
0.3

2364
R1072E
1072
−0.51
1.51
0.4
0.47
0.83

2365
R1072D
1072
−0.51
1.17
0.69
0.45
0.71

2366
D1074S
1074
1.25
1.02
0.9
1.06
0.15

2367
D1074R
1074
0.4
0.95
0.91
0.75
0.25

2368
S1075V
1075
0.52
0.36
0.53
0.47
0.08

2369
E1076T
1076
0.89
0.22
0.43
0.51
0.28

2370
K1077R
1077
0.72
0.76
0.79
0.76
0.03

2371
C1081A
1081
0.19
0.67
1.25
0.7
0.44

2372
C1081I
1081
0.7
0.77
0.29
0.59
0.21

2373
T1083E
1083
1.03
0.96
1.2
1.07
0.1

2374
T1083M
1083
0.47
1.25
1.3
1.01
0.38

2375
T1083K
1083
0.75
0.82
1.39
0.99
0.29

2376
T1083Y
1083
1.28
0.6
0.88
0.92
0.28

2377
N1087A
1087
1.01
1.13
0.6
0.91
0.23

2378
N1087W
1087
0.54
0.78
1.07
0.8
0.22

2379
N1088P
1088
2.39
0.96
0
1.12
0.98

2380
N1088A
1088
0.79
0.27
0.7
0.59
0.23

2381
I1090G
1090
0.19
1.88
1.95
1.34
0.81

2382
T1091W
1091
1.58
0.27
0.7
0.85
0.54

2383
T1091C
1091
0.37
0.74
0.59
0.57
0.15

2384
Q1092C
1092
1.8
0.96
1.8
1.52
0.39

2385
Q1092I
1092
0.88
0.27
1.1
0.75
0.35

2386
Q1092A
1092
1.17
0.56
0.29
0.67
0.37

2387
Q1092F
1092
0.59
1.04
0.29
0.64
0.31

2388
Q1092Y
1092
0.45
0.61
0.41
0.49
0.08

2389
N1093C
1093
0.05
0.67
0.76
0.49
0.31

2390
T1094I
1094
1.58
0.27
1.39
1.08
0.58

2391
T1094Y
1094
1.11
0.67
0.7
0.83
0.2

2392
T1094P
1094
−0.5
0.96
1.51
0.66
0.85

2393
V1095A
1095
0.39
0.56
0.59
0.52
0.09

2394
V1095D
1095
0.41
0.59
0.49
0.49
0.07

2395
V1095R
1095
0.27
0.61
0.42
0.43
0.14

2396
M1096C
1096
1.1
0.96
0
0.69
0.49

2397
S1097D
1097
0.67
0.64
0.8
0.7
0.07

2398
K1098W
1098
0.88
0.96
0.51
0.79
0.2

2399
K1098C
1098
0.3
0.84
0.77
0.64
0.24

2400
K1098V
1098
0.58
0.66
0.51
0.58
0.06

2401
S1099P
1099
1.02
0.72
0.45
0.73
0.23

2402
S1099Q
1099
0.32
0.77
0.69
0.59
0.19

2403
S1100D
1100
1.29
0.96
1.1
1.12
0.13

2404
S1100G
1100
0.76
1.02
1.27
1.02
0.21

2405
S1100Q
1100
0.98
0.27
0.41
0.55
0.31

2406
S1100H
1100
0.59
0.18
0.77
0.52
0.25

2407
S1100M
1100
0.38
0.52
0.62
0.51
0.1

2408
Y1106I
1106
0.43
0.42
0.43
0.43
0.01

2409
K1111R
1111
0.63
0.59
0.5
0.57
0.06

2410
K1111V
1111
0.71
0.24
0.42
0.46
0.19

2411
R1113N
1113
0.94
0.9
1.05
0.97
0.06

2412
R1113K
1113
1.13
1.12
0.63
0.96
0.23

2413
R1113I
1113
0.67
0.65
0.69
0.67
0.02

2414
R1113T
1113
0.35
0.2
1.35
0.63
0.51

2415
R1113E
1113
−0.1
0.92
0.51
0.44
0.42

2416
R1118F
1118
0.7
1.04
0.84
0.86
0.14

2417
R1118Q
1118
−0.22
1.25
0.98
0.67
0.64

2418
R1118M
1118
0.08
1.01
0.64
0.58
0.38

2419
F1119W
1119
0.47
0.87
1.5
0.95
0.42

2420
F1119A
1119
0.3
0.93
0.31
0.52
0.29

2421
F1119N
1119
0.77
0.96
−0.23
0.5
0.52

2422
S1120D
1120
0.59
0.49
0.65
0.58
0.06

2423
D1124T
1124
0.59
0.27
0.4
0.42
0.13

2424
T1125W
1125
2.27
2.41
1.1
1.92
0.59

2425
T1125D
1125
1.8
1.65
1.38
1.61
0.17

2426
T1125M
1125
0.64
0.72
1.63
1
0.45

2427
T1125H
1125
0.41
1.18
0.81
0.8
0.32

2428
T1125E
1125
0.75
0.27
0.69
0.57
0.21

2429
I1126C
1126
1.58
0.96
2.3
1.61
0.55

2430
I1126E
1126
1.58
1.37
1.61
1.52
0.11

2431
I1126A
1126
0.7
0.27
1.46
0.81
0.49

2432
T1129V
1129
1.8
1.66
1.79
1.75
0.06

2433
T1129H
1129
0.19
2.22
1.94
1.45
0.9

2434
T1129I
1129
0.88
0.78
0.69
0.78
0.08

2435
D1131L
1131
1
0.67
0.4
0.69
0.25

2436
D1131S
1131
0.51
−0.2
1.28
0.53
0.61

2437
E1133T
1133
0.19
0.67
0.4
0.42
0.2

2438
K1134I
1134
0.19
0.67
1.13
0.66
0.39

2439
T1135V
1135
0.41
0.26
1.24
0.64
0.43

2440
T1135Q
1135
0.48
0.26
0.84
0.53
0.24

2441
T1135R
1135
0.96
0.55
−0.19
0.44
0.48

2442
E1137Q
1137
0.51
0.26
0.62
0.46
0.15

2443
E1137L
1137
0.86
0.6
−0.14
0.44
0.42

2444
E1137N
1137
0.45
0.53
0.33
0.44
0.08

2445
M1138G
1138
1.89
2.46
1.38
1.91
0.44

2446
M1138D
1138
3.23
0.26
0.68
1.39
1.31

2447
T1139V
1139
0.81
1.58
0.99
1.12
0.33

2448
T1139Y
1139
0.85
0.63
0.55
0.67
0.13

2449
T1139Q
1139
0.71
0.53
0.13
0.46
0.24

2450
I1141P
1141
0.57
0.96
0.95
0.82
0.18

2451
I1141C
1141
0.59
0.38
0.48
0.48
0.09

2452
N1142A
1142
0.33
0.58
0.46
0.46
0.1

2453
W1143F
1143
0.23
0.55
0.5
0.43
0.14

2454
D1145F
1145
0.74
0.31
0.3
0.45
0.2

2455
G1146K
1146
0.52
0.53
0.35
0.47
0.08

2456
R1150N
1150
1.1
1.18
1.08
1.12
0.04

2457
Q1151C
1151
0.82
0.43
0.3
0.52
0.22

2458
D1155M
1155
0.57
0.52
0.5
0.53
0.03

2459
I1158A
1158
0.79
0.33
0.54
0.55
0.19

2460
V1159D
1159
0.77
0.43
1
0.74
0.24

2461
F1163C
1163
0.44
0.94
0.59
0.66
0.21

2462
F1163N
1163
0.5
0.37
0.53
0.47
0.07

2463
E1164P
1164
1
0.25
0.2
0.48
0.36

2464
I1165A
1165
0.53
0.37
0.46
0.45
0.07

2465
L1179D
1179
1.06
1.38
1.22
1.22
0.13

2466
L1179E
1179
0.51
0.77
0.88
0.72
0.15

2467
L1179N
1179
0.62
0.55
0.44
0.53
0.07

2468
R1182Y
1182
1.57
0.25
1.58
1.13
0.62

2469
R1182Q
1182
1.06
1.04
1
1.03
0.02

2470
R1182K
1182
0.32
1.14
0.8
0.75
0.34

2471
R1182N
1182
0.59
−0.04
0.78
0.44
0.35

2472
D1183Y
1183
0.89
1.03
0.91
0.94
0.06

2473
Y1184W
1184
0.68
0.5
0.35
0.51
0.13

2474
R1186M
1186
0.45
0.83
0.48
0.59
0.17

2475
L1187M
1187
1.65
0.01
0.46
0.71
0.69

2476
L1192Y
1192
2.13
1.34
1.35
1.61
0.37

2477
L1192K
1192
0.43
0.49
0.41
0.45
0.03

2478
E1194C
1194
0.67
0.8
1.02
0.83
0.14

2479
E1194S
1194
1.59
−0.04
−0.02
0.51
0.77

2480
E1194F
1194
0.52
0.62
0.2
0.45
0.18

2481
N1195Y
1195
1.9
0.51
0.11
0.84
0.76

2482
N1195S
1195
0.39
0.79
0.51
0.56
0.17

2483
N1196A
1196
0.74
0.61
0.52
0.62
0.09

2484
I1197W
1197
0.13
0.69
0.74
0.52
0.28

2485
I1197G
1197
0.4
0.84
0.11
0.45
0.3

2486
A1204C
1204
0.58
1.42
1.2
1.07
0.35

2487
A1204M
1204
0.76
0.67
1.13
0.86
0.2

2488
A1204Q
1204
1.16
0.53
0.39
0.69
0.33

2489
A1204F
1204
0.32
0.51
0.81
0.55
0.2

2490
A1204D
1204
0.66
0.56
0.36
0.53
0.13

2491
A1204N
1204
0.51
0.37
0.54
0.47
0.08

2492
D1206C
1206
1.22
1.02
1.05
1.1
0.09

2493
D1206I
1206
0.01
0.82
0.51
0.45
0.34

2494
D1206F
1206
0.38
0.3
0.65
0.44
0.15

2495
A1207Q
1207
0.97
0.58
0.54
0.7
0.19

2496
L1208Q
1208
0.78
0.03
0.51
0.44
0.31

2497
L1208A
1208
0.35
0.37
0.6
0.44
0.12

2498
D1211C
1211
0.9
0.61
0.74
0.75
0.12

2499
D1211T
1211
0.64
0.68
0.7
0.67
0.02

2500
D1211S
1211
0.63
0.51
0.69
0.61
0.07

2501
A1212C
1212
0.94
0.84
0.89
0.89
0.04

2502
A1212T
1212
0.8
0.74
0.7
0.75
0.04

2503
A1212S
1212
0.65
0.6
0.61
0.62
0.02

2504
A1212V
1212
0.51
0.52
0.68
0.57
0.08

2505
A1212M
1212
0.5
0.5
0.33
0.44
0.08

2506
N1215G
1215
0.62
0.65
0.91
0.73
0.13

2507
E1227T
1227
0.62
0.59
0.12
0.44
0.23

2508
I1228H
1228
1.39
0.87
0.4
0.89
0.4

2509
I1228Y
1228
0.89
0.18
0.65
0.57
0.29

2510
K1229T
1229
0.44
0.45
0.39
0.43
0.03

2511
Q1230T
1230
0.5
0.73
0.25
0.49
0.19

2512
Q1230C
1230
−0.55
0.59
1.4
0.48
0.8

2513
Q1230G
1230
0.14
1.28
−0.1
0.44
0.61

2514
E1233W
1233
0.7
1.1
1.69
1.17
0.41

2515
E1233Y
1233
0.45
0.68
0.4
0.51
0.13

2516
N1234A
1234
1.53
1.58
0.41
1.17
0.54

2517
N1234I
1234
0.43
1.04
1.29
0.92
0.36

2518
N1234E
1234
1.12
0.88
−0.51
0.5
0.72

2519
N1234F
1234
0.83
−0.22
0.75
0.45
0.48

2520
W1235Y
1235
1.64
1.69
1.15
1.5
0.24

2521
W1235E
1235
0.83
0.52
1
0.78
0.2

2522
K1236F
1236
0.54
1.03
0.74
0.77
0.2

2523
K1236P
1236
0.61
0.93
0.49
0.67
0.19

2524
E1237I
1237
0.63
0.62
0.72
0.66
0.04

2525
E1237G
1237
0.59
0.32
1.04
0.65
0.3

2526
E1237A
1237
0.51
0.68
0.54
0.58
0.07

2527
E1237F
1237
0
0.5
0.81
0.44
0.33

2528
G1239H
1239
1.24
2.06
1.15
1.48
0.41

2529
G1239K
1239
0.88
0.37
0.73
0.66
0.21

2530
G1239T
1239
0.68
0.35
0.59
0.54
0.14

2531
G1239L
1239
0.57
0.42
0.51
0.5
0.06

2532
G1239Q
1239
0.14
1.18
0.15
0.49
0.49

2533
K1240Y
1240
0.63
0.43
0.72
0.59
0.12

2534
F1241P
1241
0.97
0.98
0.49
0.81
0.23

2535
S1242E
1242
0.82
0.66
0.7
0.72
0.07

2536
S1242Q
1242
0.65
0.28
0.41
0.44
0.15

2537
D1244I
1244
0.5
0.85
0.85
0.73
0.16

2538
K1245G
1245
0.88
0.93
1.09
0.97
0.09

2539
K1245V
1245
0.53
0.43
0.54
0.5
0.05

2540
S1249E
1249
1.06
0.6
1.04
0.9
0.21

2541
S1249Q
1249
0.51
0.71
0.81
0.67
0.12

2542
S1249W
1249
0.4
0.65
0.27
0.44
0.16

2543
N1250L
1250
0.95
0.6
0.46
0.67
0.2

2544
N1250P
1250
0.58
0.63
0.46
0.56
0.07

2545
N1250G
1250
0.75
0.71
0.2
0.55
0.25

2546
N1250A
1250
0.79
0.53
0.17
0.5
0.25

2547
D1252L
1252
0.14
1.99
2.11
1.41
0.9

2548
F1254T
1254
0.83
0.19
0.3
0.44
0.28

2549
D1255M
1255
0.83
1.99
1.01
1.28
0.51

2550
F1256R
1256
0.83
0.19
0.6
0.54
0.26

2551
F1256W
1256
0.54
0.34
0.51
0.47
0.09

2552
K1260G
1260
1.06
1.71
1.78
1.51
0.33

2553
K1260F
1260
0.14
0.54
0.7
0.46
0.23

2554
K1260H
1260
0.78
0.31
0.24
0.44
0.24

2555
R1261Y
1261
1.06
0.9
1.42
1.12
0.22

2556
R1261V
1261
1.24
1.3
0.6
1.05
0.32

2557
R1261W
1261
1.59
0.76
0.32
0.89
0.53

2558
R1261N
1261
0.83
0.21
1.3
0.78
0.45

2559
R1261M
1261
1.24
0.21
0.6
0.68
0.43

2560
Y1262M
1262
0.14
0.75
0.53
0.47
0.25

2561
L1263N
1263
0.92
1.31
0.61
0.95
0.29

2562
L1263Q
1263
0.77
1.1
0.81
0.89
0.15

2563
L1263A
1263
0.81
0.87
0.51
0.73
0.16

2564
L1263E
1263
0.84
0.83
0.37
0.68
0.22

The polynucleotide and polypeptide sequences of the 2564 mutants shown in Table 2 are not provided. A person having ordinary skill in the art would understand that the mutations can be created from the wild type ERCAS12A nucleotide or polypeptide sequences by changing the codons for the wild type amino acid and changing them to a codon for the mutant amino acid. For Example, ERCAS12A mutant 1 shown in Table 2, “M1G” indicates that methionine 1 was changed to a glycine in the polypeptide sequence. Similarly, the wild type codon for methionine 1, “ATG,” can be changed to any one of the E. coli glycine codons “GGT,” GGC,” GGA,” or “GGG” to create the mutant polynucleotide coding sequence that would encode a glycine at position 1. The codons for the ERCAS12A amino acids can be optimized for a specific organism such as E. coli, Homo sapiens, or other organisms by changing the coding strand codons to codons which are most commonly used in the respective organsim.

To demonstrate the utility of these results, a set of ERCAS12A variants were selectively cloned with point mutations (135L, G39K, G39Q, 140Q, D56P, D56R, D291N, V296R, P297F, P297V, F465R, S469K, 1498K, Y507S, Y507W, Q510V, and Q510Y), and tested their activities in the context of bacteria selection assay. As shown in FIG. 2-5 most of the characterized mutants improved the survival rate of bacteria over WT-ERCAS12A (SEQ ID NO: 2) upon selection, thus validating the result of high-throughput screening. Since the survival rate in the bacterial assay is linked to the intrinsic cleavage activity of ERCAS12A nuclease, these mutants hold great promise for human genome engineering with improved performance over WT enzyme.

Table 3 shows the primer sequences used to generate an ERCAS12A saturation mutagenesis library. Standard recombinant methods and techniques were used. The screening library was constructed using the method descried in Wrenbeck et al., Nat. Methods 13(11):928-930 (2016).

The screen works by transforming a screening strain harboring a toxin plasmid with the mutant ERCAS12A library and crRNA targeting the HPRT38346 site on the toxin plasmid. After recovery and IPTG induction, cells were plated on LB-chloramphenicol media with arabinose and incubated at 37° C. overnight. Functional ERCAS12A enzymes are capable of CRISPR activity and deactivate the toxin plasmid. If the mutant LbCas12a enzyme is not functional the transformed screening E. coli cells do not survive. ERCAS12A expression plasmids carried by the surviving E. coli cells were extracted and purified. Both input and selected plasmid libraries were amplified by PCR, randomly fragmented, and sequenced. The frequencies of mutations at each position of ERCAS12A in both libraries were measured and normalized to the total coverage of each codon. The relative survival rate of each point mutation was calculated as the ratio of normalized frequency between selected and input library. Since the degree of cell survival under the arabinose selection is indicative of the cleavage activity of ERCAS12A variant at HPRT38346 protospacer, any variants that enriched during the selection over WT are those with enhanced activity at the TTTT PAM.

TABLE 3

Primer Sequences for ERCAS12A Saturation Mutagenesis Library

SEQ ID

Name
NO
DNA Sequence (5′→3′)

ERCAS12A-
211
/5Phos/taagtataagaaggagatatacatNNKAATAACGGTACCAATAACTTTCAA

P1

ERCAS12A-
212
/5Phos/gtataagaaggagatatacatatgNNKAACGGTACCAATAACTTTCAAAAT

P2

ERCAS12A-
213
/5Phos/taagaaggagatatacatatgAATNNKGGTACCAATAACTTTCAAAATTTT

P3

ERCAS12A-
214
/5Phos/gaaggagatatacatatgAATAACNNKACCAATAACTTTCAAAATTTTATC

P4

ERCAS12A-
215
/5Phos/ggagatatacatatgAATAACGGTNNKAATAACTTTCAAAATTTTATCGGG

P5

ERCAS12A-
216
/5Phos/gatatacatatgAATAACGGTACCNNKAACTTTCAAAATTTTATCGGGATC

P6

ERCAS12A-
217
/5Phos/atacatatgAATAACGGTACCAATNNKTTTCAAAATTTTATCGGGATCTCC

P7

ERCAS12A-
218
/5Phos/catatgAATAACGGTACCAATAACNNKCAAAATTTTATCGGGATCTCCTCG

P8

ERCAS12A-
219
/5Phos/atgAATAACGGTACCAATAACTTTNNKAATTTTATCGGGATCTCCTCGTTG

P9

ERCAS12A-
220
/5Phos/AATAACGGTACCAATAACTTTCAANNKTTTATCGGGATCTCCTCGTTGCAA

P10

ERCAS12A-
221
/5Phos/AACGGTACCAATAACTTTCAAAATNNKATCGGGATCTCCTCGTTGCAAAAA

P11

ERCAS12A-
222
/5Phos/GGTACCAATAACTTTCAAAATTTTNNKGGGATCTCCTCGTTGCAAAAAACT

P12

ERCAS12A-
223
/5Phos/ACCAATAACTTTCAAAATTTTATCNNKATCTCCTCGTTGCAAAAAACTCTT

P13

ERCAS12A-
224
/5Phos/AATAACTTTCAAAATTTTATCGGGNNKTCCTCGTTGCAAAAAACTCTTCGT

P14

ERCAS12A-
225
/5Phos/AACTTTCAAAATTTTATCGGGATCNNKTCGTTGCAAAAAACTCTTCGTAAT

P15

ERCAS12A-
226
/5Phos/TTTCAAAATTTTATCGGGATCTCCNNKTTGCAAAAAACTCTTCGTAATGCT

P16

ERCAS12A-
227
/5Phos/CAAAATTTTATCGGGATCTCCTCGNNKCAAAAAACTCTTCGTAATGCTTTA

P17

ERCAS12A-
228
/5Phos/AATTTTATCGGGATCTCCTCGTTGNNKAAAACTCTTCGTAATGCTTTAATC

P18

ERCAS12A-
229
/5Phos/TTTATCGGGATCTCCTCGTTGCAANNKACTCTTCGTAATGCTTTAATCCCG

P19

ERCAS12A-
230
/5Phos/ATCGGGATCTCCTCGTTGCAAAAANNKCTTCGTAATGCTTTAATCCCGACT

P20

ERCAS12A-
231
/5Phos/GGGATCTCCTCGTTGCAAAAAACTNNKCGTAATGCTTTAATCCCGACTGAG

P21

ERCAS12A-
232
/5Phos/ATCTCCTCGTTGCAAAAAACTCTTNNKAATGCTTTAATCCCGACTGAGACA

22

ERCAS12A-
233
/5Phos/TCCTCGTTGCAAAAAACTCTTCGTNNKGCTTTAATCCCGACTGAGACAACA

P23

ERCAS12A-
234
/5Phos/TCGTTGCAAAAAACTCTTCGTAATNNKTTAATCCCGACTGAGACAACACAG

P24

ERCAS12A-
235
/5Phos/TTGCAAAAAACTCTTCGTAATGCTNNKATCCCGACTGAGACAACACAGCAG

P25

ERCAS12A-
236
/5Phos/CAAAAAACTCTTCGTAATGCTTTANNKCCGACTGAGACAACACAGCAGTTC

P26

ERCAS12A-
237
/5Phos/AAAACTCTTCGTAATGCTTTAATCNNKACTGAGACAACACAGCAGTTCATC

P27

ERCAS12A-
238
/5Phos/ACTCTTCGTAATGCTTTAATCCCGNNKGAGACAACACAGCAGTTCATCGTT

P28

ERCAS12A-
239
/5Phos/CTTCGTAATGCTTTAATCCCGACTNNKACAACACAGCAGTTCATCGTTAAA

P29

ERCAS12A-
240
/5Phos/CGTAATGCTTTAATCCCGACTGAGNNKACACAGCAGTTCATCGTTAAAAAC

P30

ERCAS12A-
241
/5Phos/AATGCTTTAATCCCGACTGAGACANNKCAGCAGTTCATCGTTAAAAACGGC

P31

ERCAS12A-
242
/5Phos/GCTTTAATCCCGACTGAGACAACANNKCAGTTCATCGTTAAAAACGGCATC

P32

ERCAS12A-
243
/5Phos/TTAATCCCGACTGAGACAACACAGNNKTTCATCGTTAAAAACGGCATCATT

P33

ERCAS12A-
244
/5Phos/ATCCCGACTGAGACAACACAGCAGNNKATCGTTAAAAACGGCATCATTAAA

P34

ERCAS12A-
245
/5Phos/CCGACTGAGACAACACAGCAGTTCNNKGTTAAAAACGGCATCATTAAAGAA

P35

ERCAS12A-
246
/5Phos/ACTGAGACAACACAGCAGTTCATCNNKAAAAACGGCATCATTAAAGAAGAT

P36

ERCAS12A-
247
/5Phos/GAGACAACACAGCAGTTCATCGTTNNKAACGGCATCATTAAAGAAGATGAG

P37

ERCAS12A-
248
/5Phos/ACAACACAGCAGTTCATCGTTAAANNKGGCATCATTAAAGAAGATGAGCTG

P38

ERCAS12A-
249
/5Phos/ACACAGCAGTTCATCGTTAAAAACNNKATCATTAAAGAAGATGAGCTGCGC

P39

ERCAS12A-
250
/5Phos/CAGCAGTTCATCGTTAAAAACGGCNNKATTAAAGAAGATGAGCTGCGCGGC

P40

ERCAS12A-
251
/5Phos/CAGTTCATCGTTAAAAACGGCATCNNKAAAGAAGATGAGCTGCGCGGCGAG

P41

ERCAS12A-
252
/5Phos/TTCATCGTTAAAAACGGCATCATTNNKGAAGATGAGCTGCGCGGCGAGAAC

P42

ERCAS12A-
253
/5Phos/ATCGTTAAAAACGGCATCATTAAANNKGATGAGCTGCGCGGCGAGAACCGT

P43

ERCAS12A-
254
/5Phos/GTTAAAAACGGCATCATTAAAGAANNKGAGCTGCGCGGCGAGAACCGTCAG

P44

ERCAS12A-
255
/5Phos/AAAAACGGCATCATTAAAGAAGATNNKCTGCGCGGCGAGAACCGTCAGATT

P45

ERCAS12A-
256
/5Phos/AACGGCATCATTAAAGAAGATGAGNNKCGCGGCGAGAACCGTCAGATTTTA

P46

ERCAS12A-
257
/5Phos/GGCATCATTAAAGAAGATGAGCTGNNKGGCGAGAACCGTCAGATTTTAAAG

P47

ERCAS12A-
258
/5Phos/ATCATTAAAGAAGATGAGCTGCGCNNKGAGAACCGTCAGATTTTAAAGGAC

P48

ERCAS12A
259
/5Phos/ATTAAAGAAGATGAGCTGCGCGGCNNKAACCGTCAGATTTTAAAGGACATC

P49

ERCAS12A-
260
/5Phos/AAAGAAGATGAGCTGCGCGGCGAGNNKCGTCAGATTTTAAAGGACATCATG

P50

ERCAS12A-
261
/5Phos/GAAGATGAGCTGCGCGGCGAGAACNNKCAGATTTTAAAGGACATCATGGAC

P51

ERCAS12A
262
/5Phos/GATGAGCTGCGCGGCGAGAACCGTNNKATTTTAAAGGACATCATGGACGAC

P52

ERCAS12A-
263
/5Phos/GAGCTGCGCGGCGAGAACCGTCAGNNKTTAAAGGACATCATGGACGACTAT

P53

ERCAS12A-
264
/5Phos/CTGCGCGGCGAGAACCGTCAGATTNNKAAGGACATCATGGACGACTATTAC

P54

ERCAS12A-
265
/5Phos/CGCGGCGAGAACCGTCAGATTTTANNKGACATCATGGACGACTATTACCGT

P55

ERCAS12A-
266
/5Phos/GGCGAGAACCGTCAGATTTTAAAGNNKATCATGGACGACTATTACCGTGGG

P56

ERCAS12A-
267
/5Phos/GAGAACCGTCAGATTTTAAAGGACNNKATGGACGACTATTACCGTGGGTTC

P57

ERCAS12A-
268
/5Phos/AACCGTCAGATTTTAAAGGACATCNNKGACGACTATTACCGTGGGTTCATC

P58

ERCAS12A-
269
/5Phos/CGTCAGATTTTAAAGGACATCATGNNKGACTATTACCGTGGGTTCATCTCA

P59

ERCAS12A-
270
/5Phos/CAGATTTTAAAGGACATCATGGACNNKTATTACCGTGGGTTCATCTCAGAA

P60

ERCAS12A-
271
/5Phos/ATTTTAAAGGACATCATGGACGACNNKTACCGTGGGTTCATCTCAGAAACA

P61

ERCAS12A-
272
/5Phos/TTAAAGGACATCATGGACGACTATNNKCGTGGGTTCATCTCAGAAACATTA

P62

ERCAS12A
273
/5Phos/AAGGACATCATGGACGACTATTACNNKGGGTTCATCTCAGAAACATTAAGC

P63

ERCAS12A-
274
/5Phos/GACATCATGGACGACTATTACCGTNNKTTCATCTCAGAAACATTAAGCAGC

P64

ERCAS12A-
275
/5Phos/ATCATGGACGACTATTACCGTGGGNNKATCTCAGAAACATTAAGCAGCATC

P65

ERCAS12A-
276
/5Phos/ATGGACGACTATTACCGTGGGTTCNNKTCAGAAACATTAAGCAGCATCGAT

P66

ERCAS12A-
277
/5Phos/GACGACTATTACCGTGGGTTCATCNNKGAAACATTAAGCAGCATCGATGAT

P67

ERCAS12A-
278
/5Phos/GACTATTACCGTGGGTTCATCTCANNKACATTAAGCAGCATCGATGATATC

P68

ERCAS12A-
279
/5Phos/TATTACCGTGGGTTCATCTCAGAANNKTTAAGCAGCATCGATGATATCGAC

P69

ERCAS12A-
280
/5Phos/TACCGTGGGTTCATCTCAGAAACANNKAGCAGCATCGATGATATCGACTGG

P70

ERCAS12A-
281
/5Phos/CGTGGGTTCATCTCAGAAACATTANNKAGCATCGATGATATCGACTGGACC

P71

ERCAS12A-
282
/5Phos/GGGTTCATCTCAGAAACATTAAGCNNKATCGATGATATCGACTGGACCTCA

P72

ERCAS12A-
283
/5Phos/TTCATCTCAGAAACATTAAGCAGCNNKGATGATATCGACTGGACCTCACTG

P73

ERCAS12A-
284
/5Phos/ATCTCAGAAACATTAAGCAGCATCNNKGATATCGACTGGACCTCACTGTTT

P74

ERCAS12A-
285
/5Phos/TCAGAAACATTAAGCAGCATCGATNNKATCGACTGGACCTCACTGTTTGAG

P75

ERCAS12A-
286
/5Phos/GAAACATTAAGCAGCATCGATGATNNKGACTGGACCTCACTGTTTGAGAAG

P76

ERCAS12A-
287
/5Phos/ACATTAAGCAGCATCGATGATATCNNKTGGACCTCACTGTTTGAGAAGATG

P77

ERCAS12A-
288
/5Phos/TTAAGCAGCATCGATGATATCGACNNKACCTCACTGTTTGAGAAGATGGAG

P78

ERCAS12A-
289
/5Phos/AGCAGCATCGATGATATCGACTGGNNKTCACTGTTTGAGAAGATGGAGATT

P79

ERCAS12A-
290
/5Phos/AGCATCGATGATATCGACTGGACCNNKCTGTTTGAGAAGATGGAGATTCAA

P80

ERCAS12A-
291
/5Phos/ATCGATGATATCGACTGGACCTCANNKTTTGAGAAGATGGAGATTCAACTT

P81

ERCAS12A-
292
/5Phos/GATGATATCGACTGGACCTCACTGNNKGAGAAGATGGAGATTCAACTTAAG

P82

ERCAS12A-
293
/5Phos/GATATCGACTGGACCTCACTGTTTNNKAAGATGGAGATTCAACTTAAGAAT

P83

ERCAS12A-
294
/5Phos/ATCGACTGGACCTCACTGTTTGAGNNKATGGAGATTCAACTTAAGAATGGA

P84

ERCAS12A-
295
/5Phos/GACTGGACCTCACTGTTTGAGAAGNNKGAGATTCAACTTAAGAATGGAGAC

P85

ERCAS12A-
296
/5Phos/TGGACCTCACTGTTTGAGAAGATGNNKATTCAACTTAAGAATGGAGACAAC

P86

ERCAS12A-
297
/5Phos/ACCTCACTGTTTGAGAAGATGGAGNNKCAACTTAAGAATGGAGACAACAAA

P87

ERCAS12A-
298
/5Phos/TCACTGTTTGAGAAGATGGAGATTNNKCTTAAGAATGGAGACAACAAAGAC

P88

ERCAS12A-
299
/5Phos/CTGTTTGAGAAGATGGAGATTCAANNKAAGAATGGAGACAACAAAGACACT

P89

ERCAS12A-
300
/5Phos/TTTGAGAAGATGGAGATTCAACTTNNKAATGGAGACAACAAAGACACTTTA

P90

ERCAS12A-
301
/5Phos/GAGAAGATGGAGATTCAACTTAAGNNKGGAGACAACAAAGACACTTTAATT

P91

ERCAS12A-
302
/5Phos/AAGATGGAGATTCAACTTAAGAATNNKGACAACAAAGACACTTTAATTAAG

P92

ERCAS12A-
303
/5Phos/ATGGAGATTCAACTTAAGAATGGANNKAACAAAGACACTTTAATTAAGGAG

P93

ERCAS12A-
304
/5Phos/GAGATTCAACTTAAGAATGGAGACNNKAAAGACACTTTAATTAAGGAGCAG

P94

ERCAS12A-
305
/5Phos/ATTCAACTTAAGAATGGAGACAACNNKGACACTTTAATTAAGGAGCAGACG

P95

ERCAS12A-
306
/5Phos/CAACTTAAGAATGGAGACAACAAANNKACTTTAATTAAGGAGCAGACGGAA

P96

ERCAS12A-
307
/5Phos/CTTAAGAATGGAGACAACAAAGACNNKTTAATTAAGGAGCAGACGGAATAT

P97

ERCAS12A-
308
/5Phos/AAGAATGGAGACAACAAAGACACTNNKATTAAGGAGCAGACGGAATATCGT

P98

ERCAS12A-
309
/5Phos/AATGGAGACAACAAAGACACTTTANNKAAGGAGCAGACGGAATATCGTAAA

P99

ERCAS12A-
310
/5Phos/GGAGACAACAAAGACACTTTAATTNNKGAGCAGACGGAATATCGTAAAGCT

P100

ERCAS12A-
311
/5Phos/GACAACAAAGACACTTTAATTAAGNNKCAGACGGAATATCGTAAAGCTATC

P101

ERCAS12A-
312
/5Phos/AACAAAGACACTTTAATTAAGGAGNNKACGGAATATCGTAAAGCTATCCAT

P102

ERCAS12A-
313
/5Phos/AAAGACACTTTAATTAAGGAGCAGNNKGAATATCGTAAAGCTATCCATAAA

P103

ERCAS12A-
314
/5Phos/GACACTTTAATTAAGGAGCAGACGNNKTATCGTAAAGCTATCCATAAAAAA

P104

ERCAS12A-
315
/5Phos/ACTTTAATTAAGGAGCAGACGGAANNKCGTAAAGCTATCCATAAAAAATTT

P105

ERCAS12A-
316
/5Phos/TTAATTAAGGAGCAGACGGAATATNNKAAAGCTATCCATAAAAAATTTGCA

P106

ERCAS12A-
317
/5Phos/ATTAAGGAGCAGACGGAATATCGTNNKGCTATCCATAAAAAATTTGCAAAC

P107

ERCAS12A-
318
/5Phos/AAGGAGCAGACGGAATATCGTAAANNKATCCATAAAAAATTTGCAAACGAT

P108

ERCAS12A-
319
/5Phos/GAGCAGACGGAATATCGTAAAGCTNNKCATAAAAAATTTGCAAACGATGAC

P109

ERCAS12A-
320
/5Phos/CAGACGGAATATCGTAAAGCTATCNNKAAAAAATTTGCAAACGATGACCGT

P110

ERCAS12A-
321
/5Phos/ACGGAATATCGTAAAGCTATCCATNNKAAATTTGCAAACGATGACCGTTTC

P111

ERCAS12A-
322
/5Phos/GAATATCGTAAAGCTATCCATAAANNKTTTGCAAACGATGACCGTTTCAAA

P112

ERCAS12A-
323
/5Phos/TATCGTAAAGCTATCCATAAAAAANNKGCAAACGATGACCGTTTCAAAAAT

P113

ERCAS12A-
324
/5Phos/CGTAAAGCTATCCATAAAAAATTTNNKAACGATGACCGTTTCAAAAATATG

P114

ERCAS12A-
325
/5Phos/AAAGCTATCCATAAAAAATTTGCANNKGATGACCGTTTCAAAAATATGTTT

P115

ERCAS12A-
326
/5Phos/GCTATCCATAAAAAATTTGCAAACNNKGACCGTTTCAAAAATATGTTTTCC

P116

ERCAS12A-
327
/5Phos/ATCCATAAAAAATTTGCAAACGATNNKCGTTTCAAAAATATGTTTTCCGCA

P117

ERCAS12A-
328
/5Phos/CATAAAAAATTTGCAAACGATGACNNKTTCAAAAATATGTTTTCCGCAAAA

P118

ERCAS12A-
329
/5Phos/AAAAAATTTGCAAACGATGACCGTNNKAAAAATATGTTTTCCGCAAAATTG

P119

ERCAS12A-
330
/5Phos/AAATTTGCAAACGATGACCGTTTCNNKAATATGTTTTCCGCAAAATTGATT

P120

ERCAS12A-
331
/5Phos/TTTGCAAACGATGACCGTTTCAAANNKATGTTTTCCGCAAAATTGATTAGC

P121

ERCAS12A-
332
/5Phos/GCAAACGATGACCGTTTCAAAAATNNKTTTTCCGCAAAATTGATTAGCGAC

P122

ERCAS12A-
333
/5Phos/AACGATGACCGTTTCAAAAATATGNNKTCCGCAAAATTGATTAGCGACATT

P123

ERCAS12A-
334
/5Phos/GATGACCGTTTCAAAAATATGTTTNNKGCAAAATTGATTAGCGACATTTTA

P124

ERCAS12A-
335
/5Phos/GACCGTTTCAAAAATATGTTTTCCNNKAAATTGATTAGCGACATTTTACCT

P125

ERCAS12A-
336
/5Phos/CGTTTCAAAAATATGTTTTCCGCANNKTTGATTAGCGACATTTTACCTGAA

P126

ERCAS12A-
337
/5Phos/TTCAAAAATATGTTTTCCGCAAAANNKATTAGCGACATTTTACCTGAATTC

P127

ERCAS12A-
338
/5Phos/AAAAATATGTTTTCCGCAAAATTGNNKAGCGACATTTTACCTGAATTCGTA

P128

ERCAS12A-
339
/5Phos/AATATGTTTTCCGCAAAATTGATTNNKGACATTTTACCTGAATTCGTAATC

P129

ERCAS12A-
340
/5Phos/ATGTTTTCCGCAAAATTGATTAGCNNKATTTTACCTGAATTCGTAATCCAT

P130

ERCAS12A-
341
/5Phos/TTTTCCGCAAAATTGATTAGCGACNNKTTACCTGAATTCGTAATCCATAAC

P131

ERCAS12A-
342
/5Phos/TCCGCAAAATTGATTAGCGACATTNNKCCTGAATTCGTAATCCATAACAAC

P132

ERCAS12A-
343
/5Phos/GCAAAATTGATTAGCGACATTTTANNKGAATTCGTAATCCATAACAACAAC

P133

ERCAS12A-
344
/5Phos/AAATTGATTAGCGACATTTTACCTNNKTTCGTAATCCATAACAACAACTAT

P134

ERCAS12A-
345
/5Phos/TTGATTAGCGACATTTTACCTGAANNKGTAATCCATAACAACAACTATTCC

P135

ERCAS12A-
346
/5Phos/ATTAGCGACATTTTACCTGAATTCNNKATCCATAACAACAACTATTCCGCA

P136

ERCAS12A-
347
/5Phos/AGCGACATTTTACCTGAATTCGTANNKCATAACAACAACTATTCCGCATCT

P137

ERCAS12A-
348
/5Phos/GACATTTTACCTGAATTCGTAATCNNKAACAACAACTATTCCGCATCTGAA

P138

ERCAS12A-
349
/5Phos/ATTTTACCTGAATTCGTAATCCATNNKAACAACTATTCCGCATCTGAAAAA

P139

ERCAS12A-
350
/5Phos/TTACCTGAATTCGTAATCCATAACNNKAACTATTCCGCATCTGAAAAAGAG

P140

ERCAS12A-
351
/5Phos/CCTGAATTCGTAATCCATAACAACNNKTATTCCGCATCTGAAAAAGAGGAG

P141

ERCAS12A-
352
/5Phos/GAATTCGTAATCCATAACAACAACNNKTCCGCATCTGAAAAAGAGGAGAAG

P142

ERCAS12A
353
/5Phos/TTCGTAATCCATAACAACAACTATNNKGCATCTGAAAAAGAGGAGAAGACC

P143

ERCAS12A-
354
/5Phos/GTAATCCATAACAACAACTATTCCNNKTCTGAAAAAGAGGAGAAGACCCAA

P144

ERCAS12A-
355
/5Phos/ATCCATAACAACAACTATTCCGCANNKGAAAAAGAGGAGAAGACCCAAGTT

P145

ERCAS12A-
356
/5Phos/CATAACAACAACTATTCCGCATCTNNKAAAGAGGAGAAGACCCAAGTTATC

P146

ERCAS12A-
357
/5Phos/AACAACAACTATTCCGCATCTGAANNKGAGGAGAAGACCCAAGTTATCAAA

P147

ERCAS12A-
358
/5Phos/AACAACTATTCCGCATCTGAAAAANNKGAGAAGACCCAAGTTATCAAACTT

P148

ERCAS12A-
359
/5Phos/AACTATTCCGCATCTGAAAAAGAGNNKAAGACCCAAGTTATCAAACTTTTT

P149

ERCAS12A-
360
/5Phos/TATTCCGCATCTGAAAAAGAGGAGNNKACCCAAGTTATCAAACTTTTTTCA

P150

ERCAS12A-
361
/5Phos/TCCGCATCTGAAAAAGAGGAGAAGNNKCAAGTTATCAAACTTTTTTCACGT

P151

ERCAS12A-
362
/5Phos/GCATCTGAAAAAGAGGAGAAGACCNNKGTTATCAAACTTTTTTCACGTTTC

P152

ERCAS12A-
363
/5Phos/TCTGAAAAAGAGGAGAAGACCCAANNKATCAAACTTTTTTCACGTTTCGCG

P153

ERCAS12A-
364
/5Phos/GAAAAAGAGGAGAAGACCCAAGTTNNKAAACTTTTTTCACGTTTCGCGACA

P154

ERCAS12A-
365
/5Phos/AAAGAGGAGAAGACCCAAGTTATCNNKCTTTTTTCACGTTTCGCGACATCT

P155

ERCAS12A-
366
/5Phos/GAGGAGAAGACCCAAGTTATCAAANNKTTTTCACGTTTCGCGACATCTTTT

P156

ERCAS12A-
367
/5Phos/GAGAAGACCCAAGTTATCAAACTTNNKTCACGTTTCGCGACATCTTTTAAA

P157

ERCAS12A-
368
/5Phos/AAGACCCAAGTTATCAAACTTTTTNNKCGTTTCGCGACATCTTTTAAAGAC

P158

ERCAS12A-
369
/5Phos/ACCCAAGTTATCAAACTTTTTTCANNKTTCGCGACATCTTTTAAAGACTAC

P159

ERCAS12A-
370
/5Phos/CAAGTTATCAAACTTTTTTCACGTNNKGCGACATCTTTTAAAGACTACTTT

P160

ERCAS12A-
371
/5Phos/GTTATCAAACTTTTTTCACGTTTCNNKACATCTTTTAAAGACTACTTTAAG

P161

ERCAS12A-
372
/5Phos/ATCAAACTTTTTTCACGTTTCGCGNNKTCTTTTAAAGACTACTTTAAGAAT

P162

ERCAS12A-
373
/5Phos/AAACTTTTTTCACGTTTCGCGACANNKTTTAAAGACTACTTTAAGAATCGT

P163

ERCAS12A-
374
/5Phos/CTTTTTTCACGTTTCGCGACATCTNNKAAAGACTACTTTAAGAATCGTGCG

P164

ERCAS12A-
375
/5Phos/TTTTCACGTTTCGCGACATCTTTTNNKGACTACTTTAAGAATCGTGCGAAT

P165

ERCAS12A-
376
/5Phos/TCACGTTTCGCGACATCTTTTAAANNKTACTTTAAGAATCGTGCGAATTGT

P166

ERCAS12A-
377
/5Phos/CGTTTCGCGACATCTTTTAAAGACNNKTTTAAGAATCGTGCGAATTGTTTC

P167

ERCAS12A-
378
/5Phos/TTCGCGACATCTTTTAAAGACTACNNKAAGAATCGTGCGAATTGTTTCAGT

P168

ERCAS12A-
379
/5Phos/GCGACATCTTTTAAAGACTACTTTNNKAATCGTGCGAATTGTTTCAGTGCA

P169

ERCAS12A-
380
/5Phos/ACATCTTTTAAAGACTACTTTAAGNNKCGTGCGAATTGTTTCAGTGCAGAT

P170

ERCAS12A-
381
/5Phos/TCTTTTAAAGACTACTTTAAGAATNNKGCGAATTGTTTCAGTGCAGATGAT

P171

ERCAS12A-
382
/5Phos/TTTAAAGACTACTTTAAGAATCGTNNKAATTGTTTCAGTGCAGATGATATT

P172

ERCAS12A-
383
/5Phos/AAAGACTACTTTAAGAATCGTGCGNNKTGTTTCAGTGCAGATGATATTTCG

P173

ERCAS12A-
384
/5Phos/GACTACTTTAAGAATCGTGCGAATNNKTTCAGTGCAGATGATATTTCGTCT

P174

ERCAS12A-
385
/5Phos/TACTTTAAGAATCGTGCGAATTGTNNKAGTGCAGATGATATTTCGTCTAGT

P175

ERCAS12A-
386
/5Phos/TTTAAGAATCGTGCGAATTGTTTCNNKGCAGATGATATTTCGTCTAGTTCA

P176

ERCAS12A-
387
/5Phos/AAGAATCGTGCGAATTGTTTCAGTNNKGATGATATTTCGTCTAGTTCATGT

P177

ERCAS12A-
388
/5Phos/AATCGTGCGAATTGTTTCAGTGCANNKGATATTTCGTCTAGTTCATGTCAC

P178

ERCAS12A-
389
/5Phos/CGTGCGAATTGTTTCAGTGCAGATNNKATTTCGTCTAGTTCATGTCACCGT

P179

ERCAS12A-
390
/5Phos/GCGAATTGTTTCAGTGCAGATGATNNKTCGTCTAGTTCATGTCACCGTATC

P180

ERCAS12A-
391
/5Phos/AATTGTTTCAGTGCAGATGATATTNNKTCTAGTTCATGTCACCGTATCGTG

P181

ERCAS12A-
392
/5Phos/TGTTTCAGTGCAGATGATATTTCGNNKAGTTCATGTCACCGTATCGTGAAT

P182

ERCAS12A-
393
/5Phos/TTCAGTGCAGATGATATTTCGTCTNNKTCATGTCACCGTATCGTGAATGAC

P183

ERCAS12A-
394
/5Phos/AGTGCAGATGATATTTCGTCTAGTNNKTGTCACCGTATCGTGAATGACAAC

P184

ERCAS12A-
395
/5Phos/GCAGATGATATTTCGTCTAGTTCANNKCACCGTATCGTGAATGACAACGCT

P185

ERCAS12A-
396
/5Phos/GATGATATTTCGTCTAGTTCATGTNNKCGTATCGTGAATGACAACGCTGAG

P186

ERCAS12A-
397
/5Phos/GATATTTCGTCTAGTTCATGTCACNNKATCGTGAATGACAACGCTGAGATT

P187

ERCAS12A-
398
/5Phos/ATTTCGTCTAGTTCATGTCACCGTNNKGTGAATGACAACGCTGAGATTTTC

P188

ERCAS12A-
399
/5Phos/TCGTCTAGTTCATGTCACCGTATCNNKAATGACAACGCTGAGATTTTCTTT

P189

ERCAS12A-
400
/5Phos/TCTAGTTCATGTCACCGTATCGTGNNKGACAACGCTGAGATTTTCTTTTCT

P190

ERCAS12A-
401
/5Phos/AGTTCATGTCACCGTATCGTGAATNNKAACGCTGAGATTTTCTTTTCTAAT

P191

ERCAS12A-
402
/5Phos/TCATGTCACCGTATCGTGAATGACNNKGCTGAGATTTTCTTTTCTAATGCG

P192

ERCAS12A-
403
/5Phos/TGTCACCGTATCGTGAATGACAACNNKGAGATTTTCTTTTCTAATGCGTTG

P193

ERCAS12A-
404
/5Phos/CACCGTATCGTGAATGACAACGCTNNKATTTTCTTTTCTAATGCGTTGGTG

P194

ERCAS12A-
405
/5Phos/CGTATCGTGAATGACAACGCTGAGNNKTTCTTTTCTAATGCGTTGGTGTAT

P195

ERCAS12A-
406
/5Phos/ATCGTGAATGACAACGCTGAGATTNNKTTTTCTAATGCGTTGGTGTATCGC

P196

ERCAS12A-
407
/5Phos/GTGAATGACAACGCTGAGATTTTCNNKTCTAATGCGTTGGTGTATCGCCGC

P197

ERCAS12A-
408
/5Phos/AATGACAACGCTGAGATTTTCTTTNNKAATGCGTTGGTGTATCGCCGCATC

P198

ERCAS12A-
409
/5Phos/GACAACGCTGAGATTTTCTTTTCTNNKGCGTTGGTGTATCGCCGCATCGTA

P199

ERCAS12A-
410
/5Phos/AACGCTGAGATTTTCTTTTCTAATNNKTTGGTGTATCGCCGCATCGTAAAG

P200

ERCAS12A-
411
/5Phos/GCTGAGATTTTCTTTTCTAATGCGNNKGTGTATCGCCGCATCGTAAAGTCC

P201

ERCAS12A-
412
/5Phos/GAGATTTTCTTTTCTAATGCGTTGNNKTATCGCCGCATCGTAAAGTCCTTG

P202

ERCAS12A-
413
/5Phos/ATTTTCTTTTCTAATGCGTTGGTGNNKCGCCGCATCGTAAAGTCCTTGTCG

P203

ERCAS12A-
414
/5Phos/TTCTTTTCTAATGCGTTGGTGTATNNKCGCATCGTAAAGTCCTTGTCGAAT

P204

ERCAS12A-
415
/5Phos/TTTTCTAATGCGTTGGTGTATCGCNNKATCGTAAAGTCCTTGTCGAATGAT

P205

ERCAS12A-
416
/5Phos/TCTAATGCGTTGGTGTATCGCCGCNNKGTAAAGTCCTTGTCGAATGATGAT

P206

ERCAS12A-
417
/5Phos/AATGCGTTGGTGTATCGCCGCATCNNKAAGTCCTTGTCGAATGATGATATC

P207

ERCAS12A-
418
/5Phos/GCGTTGGTGTATCGCCGCATCGTANNKTCCTTGTCGAATGATGATATCAAC

P208

ERCAS12A-
419
/5Phos/TTGGTGTATCGCCGCATCGTAAAGNNKTTGTCGAATGATGATATCAACAAA

P209

ERCAS12A-
420
/5Phos/GTGTATCGCCGCATCGTAAAGTCCNNKTCGAATGATGATATCAACAAAATT

P210

ERCAS12A-
421
/5Phos/TATCGCCGCATCGTAAAGTCCTTGNNKAATGATGATATCAACAAAATTAGT

P211

ERCAS12A-
422
/5Phos/CGCCGCATCGTAAAGTCCTTGTCGNNKGATGATATCAACAAAATTAGTGGT

P212

ERCAS12A-
423
/5Phos/CGCATCGTAAAGTCCTTGTCGAATNNKGATATCAACAAAATTAGTGGTGAC

P213

ERCAS12A-
424
/5Phos/ATCGTAAAGTCCTTGTCGAATGATNNKATCAACAAAATTAGTGGTGACATG

P214

ERCAS12A-
425
/5Phos/GTAAAGTCCTTGTCGAATGATGATNNKAACAAAATTAGTGGTGACATGAAA

P215

ERCAS12A-
426
/5Phos/AAGTCCTTGTCGAATGATGATATCNNKAAAATTAGTGGTGACATGAAAGAC

P216

ERCAS12A-
427
/5Phos/TCCTTGTCGAATGATGATATCAACNNKATTAGTGGTGACATGAAAGACTCC

P217

ERCAS12A-
428
/5Phos/TTGTCGAATGATGATATCAACAAANNKAGTGGTGACATGAAAGACTCCCTG

P218

ERCAS12A-
429
/5Phos/TCGAATGATGATATCAACAAAATTNNKGGTGACATGAAAGACTCCCTGAAA

P219

ERCAS12A-
430
/5Phos/AATGATGATATCAACAAAATTAGTNNKGACATGAAAGACTCCCTGAAAGAA

P220

ERCAS12A-
431
/5Phos/GATGATATCAACAAAATTAGTGGTNNKATGAAAGACTCCCTGAAAGAAATG

P221

ERCAS12A-
432
/5Phos/GATATCAACAAAATTAGTGGTGACNNKAAAGACTCCCTGAAAGAAATGTCA

P222

ERCAS12A-
433
/5Phos/ATCAACAAAATTAGTGGTGACATGNNKGACTCCCTGAAAGAAATGTCATTA

P223

ERCAS12A-
434
/5Phos/AACAAAATTAGTGGTGACATGAAANNKTCCCTGAAAGAAATGTCATTAGAG

P224

ERCAS12A-
435
/5Phos/AAAATTAGTGGTGACATGAAAGACNNKCTGAAAGAAATGTCATTAGAGGAG

P225

ERCAS12A-
436
/5Phos/ATTAGTGGTGACATGAAAGACTCCNNKAAAGAAATGTCATTAGAGGAGATT

P226

ERCAS12A-
437
/5Phos/AGTGGTGACATGAAAGACTCCCTGNNKGAAATGTCATTAGAGGAGATTTAC

P227

ERCAS12A-
438
/5Phos/GGTGACATGAAAGACTCCCTGAAANNKATGTCATTAGAGGAGATTTACAGC

P228

ERCAS12A-
439
/5Phos/GACATGAAAGACTCCCTGAAAGAANNKTCATTAGAGGAGATTTACAGCTAC

P229

ERCAS12A-
440
/5Phos/ATGAAAGACTCCCTGAAAGAAATGNNKTTAGAGGAGATTTACAGCTACGAA

P230

ERCAS12A-
441
/5Phos/AAAGACTCCCTGAAAGAAATGTCANNKGAGGAGATTTACAGCTACGAAAAA

P231

ERCAS12A-
442
/5Phos/GACTCCCTGAAAGAAATGTCATTANNKGAGATTTACAGCTACGAAAAATAC

P232

ERCAS12A-
443
/5Phos/TCCCTGAAAGAAATGTCATTAGAGNNKATTTACAGCTACGAAAAATACGGA

P233

ERCAS12A-
444
/5Phos/CTGAAAGAAATGTCATTAGAGGAGNNKTACAGCTACGAAAAATACGGAGAG

P234

ERCAS12A-
445
/5Phos/AAAGAAATGTCATTAGAGGAGATTNNKAGCTACGAAAAATACGGAGAGTTC

P235

ERCAS12A-
446
/5Phos/GAAATGTCATTAGAGGAGATTTACNNKTACGAAAAATACGGAGAGTTCATC

P236

ERCAS12A-
447
/5Phos/ATGTCATTAGAGGAGATTTACAGCNNKGAAAAATACGGAGAGTTCATCACT

P237

ERCAS12A-
448
/5Phos/TCATTAGAGGAGATTTACAGCTACNNKAAATACGGAGAGTTCATCACTCAG

P238

ERCAS12A-
449
/5Phos/TTAGAGGAGATTTACAGCTACGAANNKTACGGAGAGTTCATCACTCAGGAA

P239

ERCAS12A-
450
/5Phos/GAGGAGATTTACAGCTACGAAAAANNKGGAGAGTTCATCACTCAGGAAGGT

P240

ERCAS12A-
451
/5Phos/GAGATTTACAGCTACGAAAAATACNNKGAGTTCATCACTCAGGAAGGTATC

P241

ERCAS12A-
452
/5Phos/ATTTACAGCTACGAAAAATACGGANNKTTCATCACTCAGGAAGGTATCTCC

P242

ERCAS12A-
453
/5Phos/TACAGCTACGAAAAATACGGAGAGNNKATCACTCAGGAAGGTATCTCCTTT

P243

ERCAS12A-
454
/5Phos/AGCTACGAAAAATACGGAGAGTTCNNKACTCAGGAAGGTATCTCCTTTTAC

P244

ERCAS12A-
455
/5Phos/TACGAAAAATACGGAGAGTTCATCNNKCAGGAAGGTATCTCCTTTTACAAC

P245

ERCAS12A-
456
/5Phos/GAAAAATACGGAGAGTTCATCACTNNKGAAGGTATCTCCTTTTACAACGAT

P246

ERCAS12A-
457
/5Phos/AAATACGGAGAGTTCATCACTCAGNNKGGTATCTCCTTTTACAACGATATT

P247

ERCAS12A-
458
/5Phos/TACGGAGAGTTCATCACTCAGGAANNKATCTCCTTTTACAACGATATTTGC

P248

ERCAS12A-
459
/5Phos/GGAGAGTTCATCACTCAGGAAGGTNNKTCCTTTTACAACGATATTTGCGGC

P249

ERCAS12A-
460
/5Phos/GAGTTCATCACTCAGGAAGGTATCNNKTTTTACAACGATATTTGCGGCAAA

P250

ERCAS12A-
461
/5Phos/TTCATCACTCAGGAAGGTATCTCCNNKTACAACGATATTTGCGGCAAAGTT

P251

ERCAS12A-
462
/5Phos/ATCACTCAGGAAGGTATCTCCTTTNNKAACGATATTTGCGGCAAAGTTAAT

P252

ERCAS12A-
463
/5Phos/ACTCAGGAAGGTATCTCCTTTTACNNKGATATTTGCGGCAAAGTTAATAGT

P253

ERCAS12A-
464
/5Phos/CAGGAAGGTATCTCCTTTTACAACNNKATTTGCGGCAAAGTTAATAGTTTC

P254

ERCAS12A-
465
/5Phos/GAAGGTATCTCCTTTTACAACGATNNKTGCGGCAAAGTTAATAGTTTCATG

P255

ERCAS12A-
466
/5Phos/GGTATCTCCTTTTACAACGATATTNNKGGCAAAGTTAATAGTTTCATGAAT

P256

ERCAS12A-
467
/5Phos/ATCTCCTTTTACAACGATATTTGCNNKAAAGTTAATAGTTTCATGAATCTG

P257

ERCAS12A-
468
/5Phos/TCCTTTTACAACGATATTTGCGGCNNKGTTAATAGTTTCATGAATCTGTAT

P258

ERCAS12A-
469
/5Phos/TTTTACAACGATATTTGCGGCAAANNKAATAGTTTCATGAATCTGTATTGT

P259

ERCAS12A-
470
/5Phos/TACAACGATATTTGCGGCAAAGTTNNKAGTTTCATGAATCTGTATTGTCAG

P260

ERCAS12A-
471
/5Phos/AACGATATTTGCGGCAAAGTTAATNNKTTCATGAATCTGTATTGTCAGAAG

P261

ERCAS12A-
472
/5Phos/GATATTTGCGGCAAAGTTAATAGTNNKATGAATCTGTATTGTCAGAAGAAT

P262

ERCAS12A-
473
/5Phos/ATTTGCGGCAAAGTTAATAGTTTCNNKAATCTGTATTGTCAGAAGAATAAG

P263

ERCAS12A-
474
/5Phos/TGCGGCAAAGTTAATAGTTTCATGNNKCTGTATTGTCAGAAGAATAAGGAA

P264

ERCAS12A-
475
/5Phos/GGCAAAGTTAATAGTTTCATGAATNNKTATTGTCAGAAGAATAAGGAAAAC

P265

ERCAS12A-
476
/5Phos/AAAGTTAATAGTTTCATGAATCTGNNKTGTCAGAAGAATAAGGAAAACAAG

P266

ERCAS12A-
477
/5Phos/GTTAATAGTTTCATGAATCTGTATNNKCAGAAGAATAAGGAAAACAAGAAT

P267

ERCAS12A-
478
/5Phos/AATAGTTTCATGAATCTGTATTGTNNKAAGAATAAGGAAAACAAGAATCTG

P268

ERCAS12A-
479
/5Phos/AGTTTCATGAATCTGTATTGTCAGNNKAATAAGGAAAACAAGAATCTGTAC

P269

ERCAS12A-
480
/5Phos/TTCATGAATCTGTATTGTCAGAAGNNKAAGGAAAACAAGAATCTGTACAAG

P270

ERCAS12A-
481
/5Phos/ATGAATCTGTATTGTCAGAAGAATNNKGAAAACAAGAATCTGTACAAGTTA

P271

ERCAS12A-
482
/5Phos/AATCTGTATTGTCAGAAGAATAAGNNKAACAAGAATCTGTACAAGTTACAA

P272

ERCAS12A-
483
/5Phos/CTGTATTGTCAGAAGAATAAGGAANNKAAGAATCTGTACAAGTTACAAAAG

P273

ERCAS12A-
484
/5Phos/TATTGTCAGAAGAATAAGGAAAACNNKAATCTGTACAAGTTACAAAAGCTT

P274

ERCAS12A-
485
/5Phos/TGTCAGAAGAATAAGGAAAACAAGNNKCTGTACAAGTTACAAAAGCTTCAT

P275

ERCAS12A-
486
/5Phos/CAGAAGAATAAGGAAAACAAGAATNNKTACAAGTTACAAAAGCTTCATAAA

P276

ERCAS12A-
487
/5Phos/AAGAATAAGGAAAACAAGAATCTGNNKAAGTTACAAAAGCTTCATAAACAG

P277

ERCAS12A-
488
/5Phos/AATAAGGAAAACAAGAATCTGTACNNKTTACAAAAGCTTCATAAACAGATC

P278

ERCAS12A-
489
/5Phos/AAGGAAAACAAGAATCTGTACAAGNNKCAAAAGCTTCATAAACAGATCTTG

P279

ERCAS12A-
490
/5Phos/GAAAACAAGAATCTGTACAAGTTANNKAAGCTTCATAAACAGATCTTGTGC

P280

ERCAS12A-
491
/5Phos/AACAAGAATCTGTACAAGTTACAANNKCTTCATAAACAGATCTTGTGCATC

P281

ERCAS12A-
492
/5Phos/AAGAATCTGTACAAGTTACAAAAGNNKCATAAACAGATCTTGTGCATCGCC

P282

ERCAS12A-
493
/5Phos/AATCTGTACAAGTTACAAAAGCTTNNKAAACAGATCTTGTGCATCGCCGAC

P283

ERCAS12A-
494
/5Phos/CTGTACAAGTTACAAAAGCTTCATNNKCAGATCTTGTGCATCGCCGACACG

P284

ERCAS12A-
495
/5Phos/TACAAGTTACAAAAGCTTCATAAANNKATCTTGTGCATCGCCGACACGTCT

P285

ERCAS12A-
496
/5Phos/AAGTTACAAAAGCTTCATAAACAGNNKTTGTGCATCGCCGACACGTCTTAT

P286

ERCAS12A-
497
/5Phos/TTACAAAAGCTTCATAAACAGATCNNKTGCATCGCCGACACGTCTTATGAA

P287

ERCAS12A-
498
/5Phos/CAAAAGCTTCATAAACAGATCTTGNNKATCGCCGACACGTCTTATGAAGTC

P288

ERCAS12A-
499
/5Phos/AAGCTTCATAAACAGATCTTGTGCNNKGCCGACACGTCTTATGAAGTCCCG

P289

ERCAS12A-
500
/5Phos/CTTCATAAACAGATCTTGTGCATCNNKGACACGTCTTATGAAGTCCCGTAT

P290

ERCAS12A-
501
/5Phos/CATAAACAGATCTTGTGCATCGCCNNKACGTCTTATGAAGTCCCGTATAAA

P291

ERCAS12A-
502
/5Phos/AAACAGATCTTGTGCATCGCCGACNNKTCTTATGAAGTCCCGTATAAATTT

P292

ERCAS12A-
503
/5Phos/CAGATCTTGTGCATCGCCGACACGNNKTATGAAGTCCCGTATAAATTTGAA

P293

ERCAS12A-
504
/5Phos/ATCTTGTGCATCGCCGACACGTCTNNKGAAGTCCCGTATAAATTTGAAAGC

P294

ERCAS12A-
505
/5Phos/TTGTGCATCGCCGACACGTCTTATNNKGTCCCGTATAAATTTGAAAGCGAC

P295

ERCAS12A-
506
/5Phos/TGCATCGCCGACACGTCTTATGAANNKCCGTATAAATTTGAAAGCGACGAA

P296

ERCAS12A-
507
/5Phos/ATCGCCGACACGTCTTATGAAGTCNNKTATAAATTTGAAAGCGACGAAGAA

P297

ERCAS12A-
508
/5Phos/GCCGACACGTCTTATGAAGTCCCGNNKAAATTTGAAAGCGACGAAGAAGTA

P298

ERCAS12A-
509
/5Phos/GACACGTCTTATGAAGTCCCGTATNNKTTTGAAAGCGACGAAGAAGTATAT

P299

ERCAS12A-
510
/5Phos/ACGTCTTATGAAGTCCCGTATAAANNKGAAAGCGACGAAGAAGTATATCAG

P300

ERCAS12A-
511
/5Phos/TCTTATGAAGTCCCGTATAAATTTNNKAGCGACGAAGAAGTATATCAGAGT

P301

ERCAS12A-
512
/5Phos/TATGAAGTCCCGTATAAATTTGAANNKGACGAAGAAGTATATCAGAGTGTG

P302

ERCAS12A-
513
/5Phos/GAAGTCCCGTATAAATTTGAAAGCNNKGAAGAAGTATATCAGAGTGTGAAT

P303

ERCAS12A-
514
/5Phos/GTCCCGTATAAATTTGAAAGCGACNNKGAAGTATATCAGAGTGTGAATGGA

P304

ERCAS12A-
515
/5Phos/CCGTATAAATTTGAAAGCGACGAANNKGTATATCAGAGTGTGAATGGATTC

P305

ERCAS12A-
516
/5Phos/TATAAATTTGAAAGCGACGAAGAANNKTATCAGAGTGTGAATGGATTCTTG

P306

ERCAS12A-
517
/5Phos/AAATTTGAAAGCGACGAAGAAGTANNKCAGAGTGTGAATGGATTCTTGGAC

P307

ERCAS12A-
518
/5Phos/TTTGAAAGCGACGAAGAAGTATATNNKAGTGTGAATGGATTCTTGGACAAT

P308

ERCAS12A-
519
/5Phos/GAAAGCGACGAAGAAGTATATCAGNNKGTGAATGGATTCTTGGACAATATC

P309

ERCAS12A-
520
/5Phos/AGCGACGAAGAAGTATATCAGAGTNNKAATGGATTCTTGGACAATATCTCA

P310

ERCAS12A-
521
/5Phos/GACGAAGAAGTATATCAGAGTGTGNNKGGATTCTTGGACAATATCTCATCA

P311

ERCAS12A-
522
/5Phos/GAAGAAGTATATCAGAGTGTGAATNNKTTCTTGGACAATATCTCATCAAAA

P312

ERCAS12A-
523
/5Phos/GAAGTATATCAGAGTGTGAATGGANNKTTGGACAATATCTCATCAAAACAC

P313

ERCAS12A-
524
/5Phos/GTATATCAGAGTGTGAATGGATTCNNKGACAATATCTCATCAAAACACATT

P314

ERCAS12A-
525
/5Phos/TATCAGAGTGTGAATGGATTCTTGNNKAATATCTCATCAAAACACATTGTT

P315

ERCAS12A-
526
/5Phos/CAGAGTGTGAATGGATTCTTGGACNNKATCTCATCAAAACACATTGTTGAA

P316

ERCAS12A-
527
/5Phos/AGTGTGAATGGATTCTTGGACAATNNKTCATCAAAACACATTGTTGAACGT

P317

ERCAS12A-
528
/5Phos/GTGAATGGATTCTTGGACAATATCNNKTCAAAACACATTGTTGAACGTTTG

P318

ERCAS12A-
529
/5Phos/AATGGATTCTTGGACAATATCTCANNKAAACACATTGTTGAACGTTTGCGC

P319

ERCAS12A-
530
/5Phos/GGATTCTTGGACAATATCTCATCANNKCACATTGTTGAACGTTTGCGCAAA

P320

ERCAS12A-
531
/5Phos/TTCTTGGACAATATCTCATCAAAANNKATTGTTGAACGTTTGCGCAAAATC

P321

ERCAS12A-
532
/5Phos/TTGGACAATATCTCATCAAAACACNNKGTTGAACGTTTGCGCAAAATCGGC

P322

ERCAS12A-
533
/5Phos/GACAATATCTCATCAAAACACATTNNKGAACGTTTGCGCAAAATCGGCGAC

P323

ERCAS12A-
534
/5Phos/AATATCTCATCAAAACACATTGTTNNKCGTTTGCGCAAAATCGGCGACAAC

P324

ERCAS12A-
535
/5Phos/ATCTCATCAAAACACATTGTTGAANNKTTGCGCAAAATCGGCGACAACTAT

P325

ERCAS12A-
536
/5Phos/TCATCAAAACACATTGTTGAACGTNNKCGCAAAATCGGCGACAACTATAAC

P326

ERCAS12A-
537
/5Phos/TCAAAACACATTGTTGAACGTTTGNNKAAAATCGGCGACAACTATAACGGA

P327

ERCAS12A-
538
/5Phos/AAACACATTGTTGAACGTTTGCGCNNKATCGGCGACAACTATAACGGATAC

P328

ERCAS12A-
539
/5Phos/CACATTGTTGAACGTTTGCGCAAANNKGGCGACAACTATAACGGATACAAC

P329

ERCAS12A-
540
/5Phos/ATTGTTGAACGTTTGCGCAAAATCNNKGACAACTATAACGGATACAACTTG

P330

ERCAS12A-
541
/5Phos/GTTGAACGTTTGCGCAAAATCGGCNNKAACTATAACGGATACAACTTGGAT

P331

ERCAS12A-
542
/5Phos/GAACGTTTGCGCAAAATCGGCGACNNKTATAACGGATACAACTTGGATAAA

P332

ERCAS12A-
543
/5Phos/CGTTTGCGCAAAATCGGCGACAACNNKAACGGATACAACTTGGATAAAATT

P333

ERCAS12A-
544
/5Phos/TTGCGCAAAATCGGCGACAACTATNNKGGATACAACTTGGATAAAATTTAT

P334

ERCAS12A-
545
/5Phos/CGCAAAATCGGCGACAACTATAACNNKTACAACTTGGATAAAATTTATATC

P335

ERCAS12A-
546
/5Phos/AAAATCGGCGACAACTATAACGGANNKAACTTGGATAAAATTTATATCGTA

P336

ERCAS12A-
547
/5Phos/ATCGGCGACAACTATAACGGATACNNKTTGGATAAAATTTATATCGTATCA

P337

ERCAS12A-
548
/5Phos/GGCGACAACTATAACGGATACAACNNKGATAAAATTTATATCGTATCAAAA

P338

ERCAS12A-
549
/5Phos/GACAACTATAACGGATACAACTTGNNKAAAATTTATATCGTATCAAAATTT

P339

ERCAS12A-
550
/5Phos/AACTATAACGGATACAACTTGGATNNKATTTATATCGTATCAAAATTTTAT

P340

ERCAS12A-
551
/5Phos/TATAACGGATACAACTTGGATAAANNKTATATCGTATCAAAATTTTATGAA

P341

ERCAS12A-
552
/5Phos/AACGGATACAACTTGGATAAAATTNNKATCGTATCAAAATTTTATGAATCA

P342

ERCAS12A-
553
/5Phos/GGATACAACTTGGATAAAATTTATNNKGTATCAAAATTTTATGAATCAGTA

P343

ERCAS12A-
554
/5Phos/TACAACTTGGATAAAATTTATATCNNKTCAAAATTTTATGAATCAGTATCC

P344

ERCAS12A-
555
/5Phos/AACTTGGATAAAATTTATATCGTANNKAAATTTTATGAATCAGTATCCCAA

P345

ERCAS12A-
556
/5Phos/TTGGATAAAATTTATATCGTATCANNKTTTTATGAATCAGTATCCCAAAAG

P346

ERCAS12A-
557
/5Phos/GATAAAATTTATATCGTATCAAAANNKTATGAATCAGTATCCCAAAAGACT

P347

ERCAS12A-
558
/5Phos/AAAATTTATATCGTATCAAAATTTNNKGAATCAGTATCCCAAAAGACTTAC

P348

ERCAS12A-
559
/5Phos/ATTTATATCGTATCAAAATTTTATNNKTCAGTATCCCAAAAGACTTACCGT

P349

ERCAS12A-
560
/5Phos/TATATCGTATCAAAATTTTATGAANNKGTATCCCAAAAGACTTACCGTGAT

P350

ERCAS12A-
561
/5Phos/ATCGTATCAAAATTTTATGAATCANNKTCCCAAAAGACTTACCGTGATTGG

P351

ERCAS12A-
562
/5Phos/GTATCAAAATTTTATGAATCAGTANNKCAAAAGACTTACCGTGATTGGGAG

P352

ERCAS12A-
563
/5Phos/TCAAAATTTTATGAATCAGTATCCNNKAAGACTTACCGTGATTGGGAGACT

P353

ERCAS12A-
564
/5Phos/AAATTTTATGAATCAGTATCCCAANNKACTTACCGTGATTGGGAGACTATT

P354

ERCAS12A-
565
/5Phos/TTTTATGAATCAGTATCCCAAAAGNNKTACCGTGATTGGGAGACTATTAAT

P355

ERCAS12A-
566
/5Phos/TATGAATCAGTATCCCAAAAGACTNNKCGTGATTGGGAGACTATTAATACG

P356

ERCAS12A-
567
/5Phos/GAATCAGTATCCCAAAAGACTTACNNKGATTGGGAGACTATTAATACGGCG

P357

ERCAS12A-
568
/5Phos/TCAGTATCCCAAAAGACTTACCGTNNKTGGGAGACTATTAATACGGCGTTA

P358

ERCAS12A-
569
/5Phos/GTATCCCAAAAGACTTACCGTGATNNKGAGACTATTAATACGGCGTTAGAA

P359

ERCAS12A-
570
/5Phos/TCCCAAAAGACTTACCGTGATTGGNNKACTATTAATACGGCGTTAGAAATC

P360

ERCAS12A-
571
/5Phos/CAAAAGACTTACCGTGATTGGGAGNNKATTAATACGGCGTTAGAAATCCAT

P361

ERCAS12A-
572
/5Phos/AAGACTTACCGTGATTGGGAGACTNNKAATACGGCGTTAGAAATCCATTAC

P362

ERCAS12A-
573
/5Phos/ACTTACCGTGATTGGGAGACTATTNNKACGGCGTTAGAAATCCATTACAAC

P363

ERCAS12A-
574
/5Phos/TACCGTGATTGGGAGACTATTAATNNKGCGTTAGAAATCCATTACAACAAC

P364

ERCAS12A-
575
/5Phos/CGTGATTGGGAGACTATTAATACGNNKTTAGAAATCCATTACAACAACATT

P365

ERCAS12A-
576
/5Phos/GATTGGGAGACTATTAATACGGCGNNKGAAATCCATTACAACAACATTCTG

P366

ERCAS12A-
577
/5Phos/TGGGAGACTATTAATACGGCGTTANNKATCCATTACAACAACATTCTGCCG

P367

ERCAS12A-
578
/5Phos/GAGACTATTAATACGGCGTTAGAANNKCATTACAACAACATTCTGCCGGGT

P368

ERCAS12A-
579
/5Phos/ACTATTAATACGGCGTTAGAAATCNNKTACAACAACATTCTGCCGGGTAAC

P369

ERCAS12A-
580
/5Phos/ATTAATACGGCGTTAGAAATCCATNNKAACAACATTCTGCCGGGTAACGGT

P370

ERCAS12A-
581
/5Phos/AATACGGCGTTAGAAATCCATTACNNKAACATTCTGCCGGGTAACGGTAAA

P371

ERCAS12A-
582
/5Phos/ACGGCGTTAGAAATCCATTACAACNNKATTCTGCCGGGTAACGGTAAATCG

P372

ERCAS12A-
583
/5Phos/GCGTTAGAAATCCATTACAACAACNNKCTGCCGGGTAACGGTAAATCGAAA

P373

ERCAS12A-
584
/5Phos/TTAGAAATCCATTACAACAACATTNNKCCGGGTAACGGTAAATCGAAAGCG

P374

ERCAS12A-
585
/5Phos/GAAATCCATTACAACAACATTCTGNNKGGTAACGGTAAATCGAAAGCGGAC

P375

ERCAS12A-
586
/5Phos/ATCCATTACAACAACATTCTGCCGNNKAACGGTAAATCGAAAGCGGACAAA

P376

ERCAS12A-
587
/5Phos/CATTACAACAACATTCTGCCGGGTNNKGGTAAATCGAAAGCGGACAAAGTG

P377

ERCAS12A-
588
/5Phos/TACAACAACATTCTGCCGGGTAACNNKAAATCGAAAGCGGACAAAGTGAAG

P378

ERCAS12A-
589
/5Phos/AACAACATTCTGCCGGGTAACGGTNNKTCGAAAGCGGACAAAGTGAAGAAG

P379

ERCAS12A-
590
/5Phos/AACATTCTGCCGGGTAACGGTAAANNKAAAGCGGACAAAGTGAAGAAGGCG

P380

ERCAS12A-
591
/5Phos/ATTCTGCCGGGTAACGGTAAATCGNNKGCGGACAAAGTGAAGAAGGCGGTT

P381

ERCAS12A-
592
/5Phos/CTGCCGGGTAACGGTAAATCGAAANNKGACAAAGTGAAGAAGGCGGTTAAG

P382

ERCAS12A-
593
/5Phos/CCGGGTAACGGTAAATCGAAAGCGNNKAAAGTGAAGAAGGCGGTTAAGAAC

P383

ERCAS12A-
594
/5Phos/GGTAACGGTAAATCGAAAGCGGACNNKGTGAAGAAGGCGGTTAAGAACGAT

P384

ERCAS12A-
595
/5Phos/AACGGTAAATCGAAAGCGGACAAANNKAAGAAGGCGGTTAAGAACGATCTT

P385

ERCAS12A-
596
/5Phos/GGTAAATCGAAAGCGGACAAAGTGNNKAAGGCGGTTAAGAACGATCTTCAG

P386

ERCAS12A-
597
/5Phos/AAATCGAAAGCGGACAAAGTGAAGNNKGCGGTTAAGAACGATCTTCAGAAG

P387

ERCAS12A-
598
/5Phos/TCGAAAGCGGACAAAGTGAAGAAGNNKGTTAAGAACGATCTTCAGAAGTCT

P388

ERCAS12A-
599
/5Phos/AAAGCGGACAAAGTGAAGAAGGCGNNKAAGAACGATCTTCAGAAGTCTATC

P389

ERCAS12A-
600
/5Phos/GCGGACAAAGTGAAGAAGGCGGTTNNKAACGATCTTCAGAAGTCTATCACG

P390

ERCAS12A-
601
/5Phos/GACAAAGTGAAGAAGGCGGTTAAGNNKGATCTTCAGAAGTCTATCACGGAG

P391

ERCAS12A-
602
/5Phos/AAAGTGAAGAAGGCGGTTAAGAACNNKCTTCAGAAGTCTATCACGGAGATT

P392

ERCAS12A-
603
/5Phos/GTGAAGAAGGCGGTTAAGAACGATNNKCAGAAGTCTATCACGGAGATTAAT

P393

ERCAS12A-
604
/5Phos/AAGAAGGCGGTTAAGAACGATCTTNNKAAGTCTATCACGGAGATTAATGAG

P394

ERCAS12A-
605
/5Phos/AAGGCGGTTAAGAACGATCTTCAGNNKTCTATCACGGAGATTAATGAGCTG

P395

ERCAS12A-
606
/5Phos/GCGGTTAAGAACGATCTTCAGAAGNNKATCACGGAGATTAATGAGCTGGTT

P396

ERCAS12A-
607
/5Phos/GTTAAGAACGATCTTCAGAAGTCTNNKACGGAGATTAATGAGCTGGTTTCC

P397

ERCAS12A-
608
/5Phos/AAGAACGATCTTCAGAAGTCTATCNNKGAGATTAATGAGCTGGTTTCCAAT

P398

ERCAS12A-
609
/5Phos/AACGATCTTCAGAAGTCTATCACGNNKATTAATGAGCTGGTTTCCAATTAC

P399

ERCAS12A-
610
/5Phos/GATCTTCAGAAGTCTATCACGGAGNNKAATGAGCTGGTTTCCAATTACAAG

P400

ERCAS12A-
611
/5Phos/CTTCAGAAGTCTATCACGGAGATTNNKGAGCTGGTTTCCAATTACAAGTTG

P401

ERCAS12A-
612
/5Phos/CAGAAGTCTATCACGGAGATTAATNNKCTGGTTTCCAATTACAAGTTGTGC

P402

ERCAS12A-
613
/5Phos/AAGTCTATCACGGAGATTAATGAGNNKGTTTCCAATTACAAGTTGTGCTCG

P403

ERCAS12A-
614
/5Phos/TCTATCACGGAGATTAATGAGCTGNNKTCCAATTACAAGTTGTGCTCGGAT

P404

ERCAS12A-
615
/5Phos/ATCACGGAGATTAATGAGCTGGTTNNKAATTACAAGTTGTGCTCGGATGAT

P405

ERCAS12A-
616
/5Phos/ACGGAGATTAATGAGCTGGTTTCCNNKTACAAGTTGTGCTCGGATGATAAC

P406

ERCAS12A-
617
/5Phos/GAGATTAATGAGCTGGTTTCCAATNNKAAGTTGTGCTCGGATGATAACATC

P407

ERCAS12A-
618
/5Phos/ATTAATGAGCTGGTTTCCAATTACNNKTTGTGCTCGGATGATAACATCAAA

P408

ERCAS12A-
619
/5Phos/AATGAGCTGGTTTCCAATTACAAGNNKTGCTCGGATGATAACATCAAAGCT

P409

ERCAS12A-
620
/5Phos/GAGCTGGTTTCCAATTACAAGTTGNNKTCGGATGATAACATCAAAGCTGAA

P410

ERCAS12A-
621
/5Phos/CTGGTTTCCAATTACAAGTTGTGCNNKGATGATAACATCAAAGCTGAAACC

P411

ERCAS12A-
622
/5Phos/GTTTCCAATTACAAGTTGTGCTCGNNKGATAACATCAAAGCTGAAACCTAT

P412

ERCAS12A-
623
/5Phos/TCCAATTACAAGTTGTGCTCGGATNNKAACATCAAAGCTGAAACCTATATC

P413

ERCAS12A-
624
/5Phos/AATTACAAGTTGTGCTCGGATGATNNKATCAAAGCTGAAACCTATATCCAT

P414

ERCAS12A-
625
/5Phos/TACAAGTTGTGCTCGGATGATAACNNKAAAGCTGAAACCTATATCCATGAA

P415

ERCAS12A-
626
/5Phos/AAGTTGTGCTCGGATGATAACATCNNKGCTGAAACCTATATCCATGAAATT

P416

ERCAS12A-
627
/5Phos/TTGTGCTCGGATGATAACATCAAANNKGAAACCTATATCCATGAAATTTCT

P417

ERCAS12A-
628
/5Phos/TGCTCGGATGATAACATCAAAGCTNNKACCTATATCCATGAAATTTCTCAT

P418

ERCAS12A-
629
/5Phos/TCGGATGATAACATCAAAGCTGAANNKTATATCCATGAAATTTCTCATATT

P419

ERCAS12A-
630
/5Phos/GATGATAACATCAAAGCTGAAACCNNKATCCATGAAATTTCTCATATTCTT

P420

ERCAS12A-
631
/5Phos/GATAACATCAAAGCTGAAACCTATNNKCATGAAATTTCTCATATTCTTAAT

P421

ERCAS12A-
632
/5Phos/AACATCAAAGCTGAAACCTATATCNNKGAAATTTCTCATATTCTTAATAAC

P422

ERCAS12A-
633
/5Phos/ATCAAAGCTGAAACCTATATCCATNNKATTTCTCATATTCTTAATAACTTC

P423

ERCAS12A-
634
/5Phos/AAAGCTGAAACCTATATCCATGAANNKTCTCATATTCTTAATAACTTCGAG

P424

ERCAS12A-
635
/5Phos/GCTGAAACCTATATCCATGAAATTNNKCATATTCTTAATAACTTCGAGGCA

P425

ERCAS12A-
636
/5Phos/GAAACCTATATCCATGAAATTTCTNNKATTCTTAATAACTTCGAGGCACAA

P426

ERCAS12A-
637
/5Phos/ACCTATATCCATGAAATTTCTCATNNKCTTAATAACTTCGAGGCACAAGAA

P427

ERCAS12A-
638
/5Phos/TATATCCATGAAATTTCTCATATTNNKAATAACTTCGAGGCACAAGAATTA

P428

ERCAS12A-
639
/5Phos/ATCCATGAAATTTCTCATATTCTTNNKAACTTCGAGGCACAAGAATTAAAA

P429

ERCAS12A-
640
/5Phos/CATGAAATTTCTCATATTCTTAATNNKTTCGAGGCACAAGAATTAAAATAT

P430

ERCAS12A-
641
/5Phos/GAAATTTCTCATATTCTTAATAACNNKGAGGCACAAGAATTAAAATATAAC

P431

ERCAS12A-
642
/5Phos/ATTTCTCATATTCTTAATAACTTCNNKGCACAAGAATTAAAATATAACCCA

P432

ERCAS12A-
643
/5Phos/TCTCATATTCTTAATAACTTCGAGNNKCAAGAATTAAAATATAACCCAGAA

P433

ERCAS12A-
644
/5Phos/CATATTCTTAATAACTTCGAGGCANNKGAATTAAAATATAACCCAGAAATT

P434

ERCAS12A-
645
/5Phos/ATTCTTAATAACTTCGAGGCACAANNKTTAAAATATAACCCAGAAATTCAT

P435

ERCAS12A-
646
/5Phos/CTTAATAACTTCGAGGCACAAGAANNKAAATATAACCCAGAAATTCATCTG

P436

ERCAS12A-
647
/5Phos/AATAACTTCGAGGCACAAGAATTANNKTATAACCCAGAAATTCATCTGGTT

P437

ERCAS12A-
648
/5Phos/AACTTCGAGGCACAAGAATTAAAANNKAACCCAGAAATTCATCTGGTTGAG

P438

ERCAS12A-
649
/5Phos/TTCGAGGCACAAGAATTAAAATATNNKCCAGAAATTCATCTGGTTGAGAGC

P439

ERCAS12A-
650
/5Phos/GAGGCACAAGAATTAAAATATAACNNKGAAATTCATCTGGTTGAGAGCGAA

P440

ERCAS12A-
651
/5Phos/GCACAAGAATTAAAATATAACCCANNKATTCATCTGGTTGAGAGCGAACTT

P441

ERCAS12A-
652
/5Phos/CAAGAATTAAAATATAACCCAGAANNKCATCTGGTTGAGAGCGAACTTAAA

P442

ERCAS12A-
653
/5Phos/GAATTAAAATATAACCCAGAAATTNNKCTGGTTGAGAGCGAACTTAAAGCT

P443

ERCAS12A-
654
/5Phos/TTAAAATATAACCCAGAAATTCATNNKGTTGAGAGCGAACTTAAAGCTTCA

P444

ERCAS12A-
655
/5Phos/AAATATAACCCAGAAATTCATCTGNNKGAGAGCGAACTTAAAGCTTCAGAA

P445

ERCAS12A-
656
/5Phos/TATAACCCAGAAATTCATCTGGTTNNKAGCGAACTTAAAGCTTCAGAATTA

P446

ERCAS12A-
657
/5Phos/AACCCAGAAATTCATCTGGTTGAGNNKGAACTTAAAGCTTCAGAATTAAAA

P447

ERCAS12A-
658
/5Phos/CCAGAAATTCATCTGGTTGAGAGCNNKCTTAAAGCTTCAGAATTAAAAAAT

P448

ERCAS12A-
659
/5Phos/GAAATTCATCTGGTTGAGAGCGAANNKAAAGCTTCAGAATTAAAAAATGTT

P449

ERCAS12A-
660
/5Phos/ATTCATCTGGTTGAGAGCGAACTTNNKGCTTCAGAATTAAAAAATGTTTTG

P450

ERCAS12A-
661
/5Phos/CATCTGGTTGAGAGCGAACTTAAANNKTCAGAATTAAAAAATGTTTTGGAC

P451

ERCAS12A-
662
/5Phos/CTGGTTGAGAGCGAACTTAAAGCTNNKGAATTAAAAAATGTTTTGGACGTA

P452

ERCAS12A-
663
/5Phos/GTTGAGAGCGAACTTAAAGCTTCANNKTTAAAAAATGTTTTGGACGTAATT

P453

ERCAS12A-
664
/5Phos/GAGAGCGAACTTAAAGCTTCAGAANNKAAAAATGTTTTGGACGTAATTATG

P454

ERCAS12A-
665
/5Phos/AGCGAACTTAAAGCTTCAGAATTANNKAATGTTTTGGACGTAATTATGAAC

P455

ERCAS12A-
666
/5Phos/GAACTTAAAGCTTCAGAATTAAAANNKGTTTTGGACGTAATTATGAACGCC

P456

ERCAS12A-
667
/5Phos/CTTAAAGCTTCAGAATTAAAAAATNNKTTGGACGTAATTATGAACGCCTTC

P457

ERCAS12A-
668
/5Phos/AAAGCTTCAGAATTAAAAAATGTINNKGACGTAATTATGAACGCCTTCCAT

P458

ERCAS12A-
669
/5Phos/GCTTCAGAATTAAAAAATGTTTTGNNKGTAATTATGAACGCCTTCCATTGG

P459

ERCAS12A-
670
/5Phos/TCAGAATTAAAAAATGTTTTGGACNNKATTATGAACGCCTTCCATTGGTGC

P460

ERCAS12A-
671
/5Phos/GAATTAAAAAATGTTTTGGACGTANNKATGAACGCCTTCCATTGGTGCAGC

P461

ERCAS12A-
672
/5Phos/TTAAAAAATGTTTTGGACGTAATTNNKAACGCCTTCCATTGGTGCAGCGTG

P462

ERCAS12A-
673
/5Phos/AAAAATGTTTTGGACGTAATTATGNNKGCCTTCCATTGGTGCAGCGTGTTT

P463

ERCAS12A-
674
/5Phos/AATGTTTTGGACGTAATTATGAACNNKTTCCATTGGTGCAGCGTGTTTATG

P464

ERCAS12A-
675
/5Phos/GTTTTGGACGTAATTATGAACGCCNNKCATTGGTGCAGCGTGTTTATGACA

P465

ERCAS12A-
676
/5Phos/TTGGACGTAATTATGAACGCCTTCNNKTGGTGCAGCGTGTTTATGACAGAG

P466

ERCAS12A-
677
/5Phos/GACGTAATTATGAACGCCTTCCATNNKTGCAGCGTGTTTATGACAGAGGAG

P467

ERCAS12A-
678
/5Phos/GTAATTATGAACGCCTTCCATTGGNNKAGCGTGTTTATGACAGAGGAGTTA

P468

ERCAS12A-
679
/5Phos/ATTATGAACGCCTTCCATTGGTGCNNKGTGTTTATGACAGAGGAGTTAGTA

P469

ERCAS12A-
680
/5Phos/ATGAACGCCTTCCATTGGTGCAGCNNKTTTATGACAGAGGAGTTAGTAGAT

P470

ERCAS12A-
681
/5Phos/AACGCCTTCCATTGGTGCAGCGTGNNKATGACAGAGGAGTTAGTAGATAAG

P471

ERCAS12A-
682
/5Phos/GCCTTCCATTGGTGCAGCGTGTTTNNKACAGAGGAGTTAGTAGATAAGGAT

P472

ERCAS12A-
683
/5Phos/TTCCATTGGTGCAGCGTGTTTATGNNKGAGGAGTTAGTAGATAAGGATAAC

P473

ERCAS12A-
684
/5Phos/CATTGGTGCAGCGTGTTTATGACANNKGAGTTAGTAGATAAGGATAACAAC

P474

ERCAS12A-
685
/5Phos/TGGTGCAGCGTGTTTATGACAGAGNNKTTAGTAGATAAGGATAACAACTTT

P475

ERCAS12A-
686
/5Phos/TGCAGCGTGTTTATGACAGAGGAGNNKGTAGATAAGGATAACAACTTTTAT

P476

ERCAS12A-
687
/5Phos/AGCGTGTTTATGACAGAGGAGTTANNKGATAAGGATAACAACTTTTATGCA

P477

ERCAS12A-
688
/5Phos/GTGTTTATGACAGAGGAGTTAGTANNKAAGGATAACAACTTTTATGCAGAA

P478

ERCAS12A-
689
/5Phos/TTTATGACAGAGGAGTTAGTAGATNNKGATAACAACTTTTATGCAGAATTG

P479

ERCAS12A-
690
/5Phos/ATGACAGAGGAGTTAGTAGATAAGNNKAACAACTTTTATGCAGAATTGGAA

P480

ERCAS12A-
691
/5Phos/ACAGAGGAGTTAGTAGATAAGGATNNKAACTTTTATGCAGAATTGGAAGAG

P481

ERCAS12A-
692
/5Phos/GAGGAGTTAGTAGATAAGGATAACNNKTTTTATGCAGAATTGGAAGAGATC

P482

ERCAS12A-
693
/5Phos/GAGTTAGTAGATAAGGATAACAACNNKTATGCAGAATTGGAAGAGATCTAT

P483

ERCAS12A-
694
/5Phos/TTAGTAGATAAGGATAACAACTTTNNKGCAGAATTGGAAGAGATCTATGAT

P484

ERCAS12A-
695
/5Phos/GTAGATAAGGATAACAACTTTTATNNKGAATTGGAAGAGATCTATGATGAG

P485

ERCAS12A-
696
/5Phos/GATAAGGATAACAACTTTTATGCANNKTTGGAAGAGATCTATGATGAGATC

P486

ERCAS12A-
697
/5Phos/AAGGATAACAACTTTTATGCAGAANNKGAAGAGATCTATGATGAGATCTAT

P487

ERCAS12A-
698
/5Phos/GATAACAACTTTTATGCAGAATTGNNKGAGATCTATGATGAGATCTATCCA

P488

ERCAS12A-
699
/5Phos/AACAACTTTTATGCAGAATTGGAANNKATCTATGATGAGATCTATCCAGTG

P489

ERCAS12A-
700
/5Phos/AACTTTTATGCAGAATTGGAAGAGNNKTATGATGAGATCTATCCAGTGATT

P490

ERCAS12A-
701
/5Phos/TTTTATGCAGAATTGGAAGAGATCNNKGATGAGATCTATCCAGTGATTTCA

P491

ERCAS12A-
702
/5Phos/TATGCAGAATTGGAAGAGATCTATNNKGAGATCTATCCAGTGATTTCACTG

P492

ERCAS12A-
703
/5Phos/GCAGAATTGGAAGAGATCTATGATNNKATCTATCCAGTGATTTCACTGTAT

P493

ERCAS12A-
704
/5Phos/GAATTGGAAGAGATCTATGATGAGNNKTATCCAGTGATTTCACTGTATAAC

P494

ERCAS12A-
705
/5Phos/TTGGAAGAGATCTATGATGAGATCNNKCCAGTGATTTCACTGTATAACTTA

P495

ERCAS12A-
706
/5Phos/GAAGAGATCTATGATGAGATCTATNNKGTGATTTCACTGTATAACTTAGTC

P496

ERCAS12A-
707
/5Phos/GAGATCTATGATGAGATCTATCCANNKATTTCACTGTATAACTTAGTCCGT

P497

ERCAS12A-
708
/5Phos/ATCTATGATGAGATCTATCCAGTGNNKTCACTGTATAACTTAGTCCGTAAT

P498

ERCAS12A-
709
/5Phos/TATGATGAGATCTATCCAGTGATTNNKCTGTATAACTTAGTCCGTAATTAC

P499

ERCAS12A-
710
/5Phos/GATGAGATCTATCCAGTGATTTCANNKTATAACTTAGTCCGTAATTACGTC

P500

ERCAS12A-
711
/5Phos/GAGATCTATCCAGTGATTTCACTGNNKAACTTAGTCCGTAATTACGTCACT

P501

ERCAS12A-
712
/5Phos/ATCTATCCAGTGATTTCACTGTATNNKTTAGTCCGTAATTACGTCACTCAG

P502

ERCAS12A-
713
/5Phos/TATCCAGTGATTTCACTGTATAACNNKGTCCGTAATTACGTCACTCAGAAG

P503

ERCAS12A-
714
/5Phos/CCAGTGATTTCACTGTATAACTTANNKCGTAATTACGTCACTCAGAAGCCG

P504

ERCAS12A-
715
/5Phos/GTGATTTCACTGTATAACTTAGTCNNKAATTACGTCACTCAGAAGCCGTAC

P505

ERCAS12A-
716
/5Phos/ATTTCACTGTATAACTTAGTCCGTNNKTACGTCACTCAGAAGCCGTACTCA

P506

ERCAS12A-
717
/5Phos/TCACTGTATAACTTAGTCCGTAATNNKGTCACTCAGAAGCCGTACTCAACT

P507

ERCAS12A-
718
/5Phos/CTGTATAACTTAGTCCGTAATTACNNKACTCAGAAGCCGTACTCAACTAAA

P508

ERCAS12A-
719
/5Phos/TATAACTTAGTCCGTAATTACGTCNNKCAGAAGCCGTACTCAACTAAAAAG

P509

ERCAS12A-
720
/5Phos/AACTTAGTCCGTAATTACGTCACTNNKAAGCCGTACTCAACTAAAAAGATC

P510

ERCAS12A-
721
/5Phos/TTAGTCCGTAATTACGTCACTCAGNNKCCGTACTCAACTAAAAAGATCAAG

P511

ERCAS12A-
722
/5Phos/GTCCGTAATTACGTCACTCAGAAGNNKTACTCAACTAAAAAGATCAAGTTA

P512

ERCAS12A-
723
/5Phos/CGTAATTACGTCACTCAGAAGCCGNNKTCAACTAAAAAGATCAAGTTAAAT

P513

ERCAS12A-
724
/5Phos/AATTACGTCACTCAGAAGCCGTACNNKACTAAAAAGATCAAGTTAAATTTT

P514

ERCAS12A-
725
/5Phos/TACGTCACTCAGAAGCCGTACTCANNKAAAAAGATCAAGTTAAATTTTGGA

P515

ERCAS12A-
726
/5Phos/GTCACTCAGAAGCCGTACTCAACTNNKAAGATCAAGTTAAATTTTGGAATT

P516

ERCAS12A-
727
/5Phos/ACTCAGAAGCCGTACTCAACTAAANNKATCAAGTTAAATTTTGGAATTCCG

P517

ERCAS12A-
728
/5Phos/CAGAAGCCGTACTCAACTAAAAAGNNKAAGTTAAATTTTGGAATTCCGACA

P518

ERCAS12A-
729
/5Phos/AAGCCGTACTCAACTAAAAAGATCNNKTTAAATTTTGGAATTCCGACATTG

P519

ERCAS12A-
730
/5Phos/CCGTACTCAACTAAAAAGATCAAGNNKAATTTTGGAATTCCGACATTGGCG

P520

ERCAS12A-
731
/5Phos/TACTCAACTAAAAAGATCAAGTTANNKTTTGGAATTCCGACATTGGCGGAT

P521

ERCAS12A-
732
/5Phos/TCAACTAAAAAGATCAAGTTAAATNNKGGAATTCCGACATTGGCGGATGGT

P522

ERCAS12A-
733
/5Phos/ACTAAAAAGATCAAGTTAAATTTTNNKATTCCGACATTGGCGGATGGTTGG

P523

ERCAS12A-
734
/5Phos/AAAAAGATCAAGTTAAATTTTGGANNKCCGACATTGGCGGATGGTTGGAGT

P524

ERCAS12A-
735
/5Phos/AAGATCAAGTTAAATTTTGGAATTNNKACATTGGCGGATGGTTGGAGTAAG

P525

ERCAS12A-
736
/5Phos/ATCAAGTTAAATTTTGGAATTCCGNNKTTGGCGGATGGTTGGAGTAAGTCT

P526

ERCAS12A-
737
/5Phos/AAGTTAAATTTTGGAATTCCGACANNKGCGGATGGTTGGAGTAAGTCTAAG

P527

ERCAS12A-
738
/5Phos/TTAAATTTTGGAATTCCGACATTGNNKGATGGTTGGAGTAAGTCTAAGGAA

P528

ERCAS12A-
739
/5Phos/AATTTTGGAATTCCGACATTGGCGNNKGGTTGGAGTAAGTCTAAGGAATAC

P529

ERCAS12A-
740
/5Phos/TTTGGAATTCCGACATTGGCGGATNNKTGGAGTAAGTCTAAGGAATACTCA

P530

ERCAS12A-
741
/5Phos/GGAATTCCGACATTGGCGGATGGTNNKAGTAAGTCTAAGGAATACTCAAAT

P531

ERCAS12A-
742
/5Phos/ATTCCGACATTGGCGGATGGTTGGNNKAAGTCTAAGGAATACTCAAATAAC

P532

ERCAS12A-
743
/5Phos/CCGACATTGGCGGATGGTTGGAGTNNKTCTAAGGAATACTCAAATAACGCT

P533

ERCAS12A-
744
/5Phos/ACATTGGCGGATGGTTGGAGTAAGNNKAAGGAATACTCAAATAACGCTATT

P534

ERCAS12A-
745
/5Phos/TTGGCGGATGGTTGGAGTAAGTCTNNKGAATACTCAAATAACGCTATTATC

P535

ERCAS12A-
746
/5Phos/GCGGATGGTTGGAGTAAGTCTAAGNNKTACTCAAATAACGCTATTATCTTA

P536

ERCAS12A-
747
/5Phos/GATGGTTGGAGTAAGTCTAAGGAANNKTCAAATAACGCTATTATCTTAATG

P537

ERCAS12A-
748
/5Phos/GGTTGGAGTAAGTCTAAGGAATACNNKAATAACGCTATTATCTTAATGCGT

P538

ERCAS12A-
749
/5Phos/TGGAGTAAGTCTAAGGAATACTCANNKAACGCTATTATCTTAATGCGTGAC

P539

ERCAS12A-
750
/5Phos/AGTAAGTCTAAGGAATACTCAAATNNKGCTATTATCTTAATGCGTGACAAC

P540

ERCAS12A-
751
/5Phos/AAGTCTAAGGAATACTCAAATAACNNKATTATCTTAATGCGTGACAACCTG

P541

ERCAS12A-
752
/5Phos/TCTAAGGAATACTCAAATAACGCTNNKATCTTAATGCGTGACAACCTGTAT

P542

ERCAS12A-
753
/5Phos/AAGGAATACTCAAATAACGCTATTNNKTTAATGCGTGACAACCTGTATTAC

P543

ERCAS12A-
754
/5Phos/GAATACTCAAATAACGCTATTATCNNKATGCGTGACAACCTGTATTACTTG

P544

ERCAS12A-
755
/5Phos/TACTCAAATAACGCTATTATCTTANNKCGTGACAACCTGTATTACTTGGGT

P545

ERCAS12A-
756
/5Phos/TCAAATAACGCTATTATCTTAATGNNKGACAACCTGTATTACTTGGGTATT

P546

ERCAS12A-
757
/5Phos/AATAACGCTATTATCTTAATGCGTNNKAACCTGTATTACTTGGGTATTTTC

P547

ERCAS12A-
758
/5Phos/AACGCTATTATCTTAATGCGTGACNNKCTGTATTACTTGGGTATTTTCAAT

P548

ERCAS12A-
759
/5Phos/GCTATTATCTTAATGCGTGACAACNNKTATTACTTGGGTATTTTCAATGCC

P549

ERCAS12A-
760
/5Phos/ATTATCTTAATGCGTGACAACCTGNNKTACTTGGGTATTTTCAATGCCAAA

P550

ERCAS12A-
761
/5Phos/ATCTTAATGCGTGACAACCTGTATNNKTTGGGTATTTTCAATGCCAAAAAT

P551

ERCAS12A-
762
/5Phos/TTAATGCGTGACAACCTGTATTACNNKGGTATTTTCAATGCCAAAAATAAG

P552

ERCAS12A-
763
/5Phos/ATGCGTGACAACCTGTATTACTTGNNKATTTTCAATGCCAAAAATAAGCCC

P553

ERCAS12A-
764
/5Phos/CGTGACAACCTGTATTACTTGGGTNNKTTCAATGCCAAAAATAAGCCCGAT

P554

ERCAS12A-
765
/5Phos/GACAACCTGTATTACTTGGGTATTNNKAATGCCAAAAATAAGCCCGATAAA

P555

ERCAS12A-
766
/5Phos/AACCTGTATTACTTGGGTATTTTCNNKGCCAAAAATAAGCCCGATAAAAAA

P556

ERCAS12A-
767
/5Phos/CTGTATTACTTGGGTATTTTCAATNNKAAAAATAAGCCCGATAAAAAAATC

P557

ERCAS12A-
768
/5Phos/TATTACTTGGGTATTTTCAATGCCNNKAATAAGCCCGATAAAAAAATCATT

P558

ERCAS12A-
769
/5Phos/TACTTGGGTATTTTCAATGCCAAANNKAAGCCCGATAAAAAAATCATTGAA

P559

ERCAS12A-
770
/5Phos/TTGGGTATTTTCAATGCCAAAAATNNKCCCGATAAAAAAATCATTGAAGGG

P560

ERCAS12A-
771
/5Phos/GGTATTTTCAATGCCAAAAATAAGNNKGATAAAAAAATCATTGAAGGGAAT

P561

ERCAS12A-
772
/5Phos/ATTTTCAATGCCAAAAATAAGCCCNNKAAAAAAATCATTGAAGGGAATACT

P562

ERCAS12A-
773
/5Phos/TTCAATGCCAAAAATAAGCCCGATNNKAAAATCATTGAAGGGAATACTTCA

P563

ERCAS12A-
774
/5Phos/AATGCCAAAAATAAGCCCGATAAANNKATCATTGAAGGGAATACTTCAGAA

P564

ERCAS12A-
775
/5Phos/GCCAAAAATAAGCCCGATAAAAAANNKATTGAAGGGAATACTTCAGAAAAT

P565

ERCAS12A-
776
/5Phos/AAAAATAAGCCCGATAAAAAAATCNNKGAAGGGAATACTTCAGAAAATAAG

P566

ERCAS12A-
777
/5Phos/AATAAGCCCGATAAAAAAATCATTNNKGGGAATACTTCAGAAAATAAGGGA

P567

ERCAS12A-
778
/5Phos/AAGCCCGATAAAAAAATCATTGAANNKAATACTTCAGAAAATAAGGGAGAT

P568

ERCAS12A-
779
/5Phos/CCCGATAAAAAAATCATTGAAGGGNNKACTTCAGAAAATAAGGGAGATTAC

P569

ERCAS12A-
780
/5Phos/GATAAAAAAATCATTGAAGGGAATNNKTCAGAAAATAAGGGAGATTACAAA

P570

ERCAS12A-
781
/5Phos/AAAAAAATCATTGAAGGGAATACTNNKGAAAATAAGGGAGATTACAAAAAA

P571

ERCAS12A
782
/5Phos/AAAATCATTGAAGGGAATACTTCANNKAATAAGGGAGATTACAAAAAAATG

P572

ERCAS12A-
783
/5Phos/ATCATTGAAGGGAATACTTCAGAANNKAAGGGAGATTACAAAAAAATGATT

P573

ERCAS12A-
784
/5Phos/ATTGAAGGGAATACTTCAGAAAATNNKGGAGATTACAAAAAAATGATTTAT

P574

ERCAS12A-
785
/5Phos/GAAGGGAATACTTCAGAAAATAAGNNKGATTACAAAAAAATGATTTATAAC

P575

ERCAS12A-
786
/5Phos/GGGAATACTTCAGAAAATAAGGGANNKTACAAAAAAATGATTTATAACCTT

P576

ERCAS12A-
787
/5Phos/AATACTTCAGAAAATAAGGGAGATNNKAAAAAAATGATTTATAACCTTTTG

P577

ERCAS12A-
788
/5Phos/ACTTCAGAAAATAAGGGAGATTACNNKAAAATGATTTATAACCTTTTGCCA

P578

ERCAS12A-
789
/5Phos/TCAGAAAATAAGGGAGATTACAAANNKATGATTTATAACCTTTTGCCAGGC

P579

ERCAS12A-
790
/5Phos/GAAAATAAGGGAGATTACAAAAAANNKATTTATAACCTTTTGCCAGGCCCA

P580

ERCAS12A
791
/5Phos/AATAAGGGAGATTACAAAAAAATGNNKTATAACCTTTTGCCAGGCCCAAAC

P581

ERCAS12A-
792
/5Phos/AAGGGAGATTACAAAAAAATGATTNNKAACCTTTTGCCAGGCCCAAACAAA

P582

ERCAS12A-
793
/5Phos/GGAGATTACAAAAAAATGATTTATNNKCTTTTGCCAGGCCCAAACAAAATG

P583

ERCAS12A-
794
/5Phos/GATTACAAAAAAATGATTTATAACNNKTTGCCAGGCCCAAACAAAATGATC

P584

ERCAS12A-
795
/5Phos/TACAAAAAAATGATTTATAACCTTNNKCCAGGCCCAAACAAAATGATCCCC

P585

ERCAS12A-
796
/5Phos/AAAAAAATGATTTATAACCTTTTGNNKGGCCCAAACAAAATGATCCCCAAG

P586

ERCAS12A-
797
/5Phos/AAAATGATTTATAACCTTTTGCCANNKCCAAACAAAATGATCCCCAAGGTG

P587

ERCAS12A-
798
/5Phos/ATGATTTATAACCTTTTGCCAGGCNNKAACAAAATGATCCCCAAGGTGTTT

P588

ERCAS12A-
799
/5Phos/ATTTATAACCTTTTGCCAGGCCCANNKAAAATGATCCCCAAGGTGTTTTTG

P589

ERCAS12A-
800
/5Phos/TATAACCTTTTGCCAGGCCCAAACNNKATGATCCCCAAGGTGTTTTTGAGT

P590

ERCAS12A-
801
/5Phos/AACCTTTTGCCAGGCCCAAACAAANNKATCCCCAAGGTGTTTTTGAGTAGT

P591

ERCAS12A-
802
/5Phos/CTTTTGCCAGGCCCAAACAAAATGNNKCCCAAGGTGTTTTTGAGTAGTAAG

P592

ERCAS12A-
803
/5Phos/TTGCCAGGCCCAAACAAAATGATCNNKAAGGTGTTTTTGAGTAGTAAGACG

P593

ERCAS12A-
804
/5Phos/CCAGGCCCAAACAAAATGATCCCCNNKGTGTTTTTGAGTAGTAAGACGGGA

P594

ERCAS12A-
805
/5Phos/GGCCCAAACAAAATGATCCCCAAGNNKTTTTTGAGTAGTAAGACGGGAGTA

P595

ERCAS12A-
806
/5Phos/CCAAACAAAATGATCCCCAAGGTGNNKTTGAGTAGTAAGACGGGAGTAGAA

P596

ERCAS12A-
807
/5Phos/AACAAAATGATCCCCAAGGTGTTTNNKAGTAGTAAGACGGGAGTAGAAACA

P597

ERCAS12A-
808
/5Phos/AAAATGATCCCCAAGGTGTTTTTGNNKAGTAAGACGGGAGTAGAAACATAT

P598

ERCAS12A-
809
/5Phos/ATGATCCCCAAGGTGTTTTTGAGTNNKAAGACGGGAGTAGAAACATATAAG

P599

ERCAS12A-
810
/5Phos/ATCCCCAAGGTGTTTTTGAGTAGTNNKACGGGAGTAGAAACATATAAGCCA

P600

ERCAS12A-
811
/5Phos/CCCAAGGTGTTTTTGAGTAGTAAGNNKGGAGTAGAAACATATAAGCCATCA

P601

ERCAS12A-
812
/5Phos/AAGGTGTTTTTGAGTAGTAAGACGNNKGTAGAAACATATAAGCCATCAGCG

P602

ERCAS12A-
813
/5Phos/GTGTTTTTGAGTAGTAAGACGGGANNKGAAACATATAAGCCATCAGCGTAC

P603

ERCAS12A-
814
/5Phos/TTTTTGAGTAGTAAGACGGGAGTANNKACATATAAGCCATCAGCGTACATT

P604

ERCAS12A-
815
/5Phos/TTGAGTAGTAAGACGGGAGTAGAANNKTATAAGCCATCAGCGTACATTCTT

P605

ERCAS12A-
816
/5Phos/AGTAGTAAGACGGGAGTAGAAACANNKAAGCCATCAGCGTACATTCTTGAA

P606

ERCAS12A-
817
/5Phos/AGTAAGACGGGAGTAGAAACATATNNKCCATCAGCGTACATTCTTGAAGGG

P607

ERCAS12A-
818
/5Phos/AAGACGGGAGTAGAAACATATAAGNNKTCAGCGTACATTCTTGAAGGGTAC

P608

ERCAS12A-
819
/5Phos/ACGGGAGTAGAAACATATAAGCCANNKGCGTACATTCTTGAAGGGTACAAA

P609

ERCAS12A-
820
/5Phos/GGAGTAGAAACATATAAGCCATCANNKTACATTCTTGAAGGGTACAAACAA

P610

ERCAS12A-
821
/5Phos/GTAGAAACATATAAGCCATCAGCGNNKATTCTTGAAGGGTACAAACAAAAT

P611

ERCAS12A-
822
/5Phos/GAAACATATAAGCCATCAGCGTACNNKCTTGAAGGGTACAAACAAAATAAA

P612

ERCAS12A-
823
/5Phos/ACATATAAGCCATCAGCGTACATTNNKGAAGGGTACAAACAAAATAAACAC

P613

ERCAS12A-
824
/5Phos/TATAAGCCATCAGCGTACATTCTTNNKGGGTACAAACAAAATAAACACATC

P614

ERCAS12A-
825
/5Phos/AAGCCATCAGCGTACATTCTTGAANNKTACAAACAAAATAAACACATCAAG

P615

ERCAS12A-
826
/5Phos/CCATCAGCGTACATTCTTGAAGGGNNKAAACAAAATAAACACATCAAGAGC

P616

ERCAS12A-
827
/5Phos/TCAGCGTACATTCTTGAAGGGTACNNKCAAAATAAACACATCAAGAGCAGC

P617

ERCAS12A-
828
/5Phos/GCGTACATTCTTGAAGGGTACAAANNKAATAAACACATCAAGAGCAGCAAG

P618

ERCAS12A-
829
/5Phos/TACATTCTTGAAGGGTACAAACAANNKAAACACATCAAGAGCAGCAAGGAC

P619

ERCAS12A-
830
/5Phos/ATTCTTGAAGGGTACAAACAAAATNNKCACATCAAGAGCAGCAAGGACTTT

P620

ERCAS12A-
831
/5Phos/CTTGAAGGGTACAAACAAAATAAANNKATCAAGAGCAGCAAGGACTTTGAC

P621

ERCAS12A-
832
/5Phos/GAAGGGTACAAACAAAATAAACACNNKAAGAGCAGCAAGGACTTTGACATT

P622

ERCAS12A-
833
/5Phos/GGGTACAAACAAAATAAACACATCNNKAGCAGCAAGGACTTTGACATTACA

P623

ERCAS12A-
834
/5Phos/TACAAACAAAATAAACACATCAAGNNKAGCAAGGACTTTGACATTACATTT

P624

ERCAS12A-
835
/5Phos/AAACAAAATAAACACATCAAGAGCNNKAAGGACTTTGACATTACATTTTGT

P625

ERCAS12A-
836
/5Phos/CAAAATAAACACATCAAGAGCAGCNNKGACTTTGACATTACATTTTGTCAT

P626

ERCAS12A-
837
/5Phos/AATAAACACATCAAGAGCAGCAAGNNKTTTGACATTACATTTTGTCATGAC

P627

ERCAS12A-
838
/5Phos/AAACACATCAAGAGCAGCAAGGACNNKGACATTACATTTTGTCATGACTTA

P628

ERCAS12A-
839
/5Phos/CACATCAAGAGCAGCAAGGACTTTNNKATTACATTTTGTCATGACTTAATC

P629

ERCAS12A-
840
/5Phos/ATCAAGAGCAGCAAGGACTTTGACNNKACATTTTGTCATGACTTAATCGAC

P630

ERCAS12A-
841
/5Phos/AAGAGCAGCAAGGACTTTGACATTNNKTTTTGTCATGACTTAATCGACTAC

P631

ERCAS12A-
842
/5Phos/AGCAGCAAGGACTTTGACATTACANNKTGTCATGACTTAATCGACTACTTT

P632

ERCAS12A-
843
/5Phos/AGCAAGGACTTTGACATTACATTTNNKCATGACTTAATCGACTACTTTAAA

P633

ERCAS12A-
844
/5Phos/AAGGACTTTGACATTACATTTTGTNNKGACTTAATCGACTACTTTAAAAAT

P634

ERCAS12A-
845
/5Phos/GACTTTGACATTACATTTTGTCATNNKTTAATCGACTACTTTAAAAATTGC

P635

ERCAS12A-
846
/5Phos/TTTGACATTACATTTTGTCATGACNNKATCGACTACTTTAAAAATTGCATT

P636

ERCAS12A-
847
/5Phos/GACATTACATTTTGTCATGACTTANNKGACTACTTTAAAAATTGCATTGCC

P637

ERCAS12A-
848
/5Phos/ATTACATTTTGTCATGACTTAATCNNKTACTTTAAAAATTGCATTGCCATT

P638

ERCAS12A-
849
/5Phos/ACATTTTGTCATGACTTAATCGACNNKTTTAAAAATTGCATTGCCATTCAC

P639

ERCAS12A-
850
/5Phos/TTTTGTCATGACTTAATCGACTACNNKAAAAATTGCATTGCCATTCACCCT

P640

ERCAS12A-
851
/5Phos/TGTCATGACTTAATCGACTACTTTNNKAATTGCATTGCCATTCACCCTGAG

P641

ERCAS12A-
852
/5Phos/CATGACTTAATCGACTACTTTAAANNKTGCATTGCCATTCACCCTGAGTGG

P642

ERCAS12A-
853
/5Phos/GACTTAATCGACTACTTTAAAAATNNKATTGCCATTCACCCTGAGTGGAAA

P643

ERCAS12A-
854
/5Phos/TTAATCGACTACTTTAAAAATTGCNNKGCCATTCACCCTGAGTGGAAAAAT

P644

ERCAS12A-
855
/5Phos/ATCGACTACTTTAAAAATTGCATTNNKATTCACCCTGAGTGGAAAAATTTT

P645

ERCAS12A-
856
/5Phos/GACTACTTTAAAAATTGCATTGCCNNKCACCCTGAGTGGAAAAATTTTGGT

P646

ERCAS12A-
857
/5Phos/TACTTTAAAAATTGCATTGCCATTNNKCCTGAGTGGAAAAATTTTGGTTTC

P647

ERCAS12A-
858
/5Phos/TTTAAAAATTGCATTGCCATTCACNNKGAGTGGAAAAATTTTGGTTTCGAC

P648

ERCAS12A-
859
/5Phos/AAAAATTGCATTGCCATTCACCCTNNKTGGAAAAATTTTGGTTTCGACTTC

P649

ERCAS12A-
860
/5Phos/AATTGCATTGCCATTCACCCTGAGNNKAAAAATTTTGGTTTCGACTTCAGT

P650

ERCAS12A-
861
/5Phos/TGCATTGCCATTCACCCTGAGTGGNNKAATTTTGGTTTCGACTTCAGTGAT

P651

ERCAS12A-
862
/5Phos/ATTGCCATTCACCCTGAGTGGAAANNKTTTGGTTTCGACTTCAGTGATACC

P652

ERCAS12A-
863
/5Phos/GCCATTCACCCTGAGTGGAAAAATNNKGGTTTCGACTTCAGTGATACCTCT

P653

ERCAS12A-
864
/5Phos/ATTCACCCTGAGTGGAAAAATTTTNNKTTCGACTTCAGTGATACCTCTACC

P654

ERCAS12A-
865
/5Phos/CACCCTGAGTGGAAAAATTTTGGTNNKGACTTCAGTGATACCTCTACCTAT

P655

ERCAS12A-
866
/5Phos/CCTGAGTGGAAAAATTTTGGTTTCNNKTTCAGTGATACCTCTACCTATGAA

P656

ERCAS12A-
867
/5Phos/GAGTGGAAAAATTTTGGTTTCGACNNKAGTGATACCTCTACCTATGAAGAT

P657

ERCAS12A-
868
/5Phos/TGGAAAAATTTTGGTTTCGACTTCNNKGATACCTCTACCTATGAAGATATC

P658

ERCAS12A-
869
/5Phos/AAAAATTTTGGTTTCGACTTCAGTNNKACCTCTACCTATGAAGATATCTCT

P659

ERCAS12A-
870
/5Phos/AATTTTGGTTTCGACTTCAGTGATNNKTCTACCTATGAAGATATCTCTGGT

P660

ERCAS12A-
871
/5Phos/TTTGGTTTCGACTTCAGTGATACCNNKACCTATGAAGATATCTCTGGTTTT

P661

ERCAS12A-
872
/5Phos/GGTTTCGACTTCAGTGATACCTCTNNKTATGAAGATATCTCTGGTTTTTAT

P662

ERCAS12A-
873
/5Phos/TTCGACTTCAGTGATACCTCTACCNNKGAAGATATCTCTGGTTTTTATCGT

P663

ERCAS12A-
874
/5Phos/GACTTCAGTGATACCTCTACCTATNNKGATATCTCTGGTTTTTATCGTGAG

P664

ERCAS12A-
875
/5Phos/TTCAGTGATACCTCTACCTATGAANNKATCTCTGGTTTTTATCGTGAGGTA

P665

ERCAS12A-
876
/5Phos/AGTGATACCTCTACCTATGAAGATNNKTCTGGTTTTTATCGTGAGGTAGAA

P666

ERCAS12A-
877
/5Phos/GATACCTCTACCTATGAAGATATCNNKGGTTTTTATCGTGAGGTAGAACTT

P667

ERCAS12A-
878
/5Phos/ACCTCTACCTATGAAGATATCTCTNNKTTTTATCGTGAGGTAGAACTTCAG

P668

ERCAS12A-
879
/5Phos/TCTACCTATGAAGATATCTCTGGTNNKTATCGTGAGGTAGAACTTCAGGGT

P669

ERCAS12A-
880
/5Phos/ACCTATGAAGATATCTCTGGTTTTNNKCGTGAGGTAGAACTTCAGGGTTAC

P670

ERCAS12A-
881
/5Phos/TATGAAGATATCTCTGGTTTTTATNNKGAGGTAGAACTTCAGGGTTACAAG

P671

ERCAS12A-
882
/5Phos/GAAGATATCTCTGGTTTTTATCGTNNKGTAGAACTTCAGGGTTACAAGATC

P672

ERCAS12A-
883
/5Phos/GATATCTCTGGTTTTTATCGTGAGNNKGAACTTCAGGGTTACAAGATCGAT

P673

ERCAS12A-
884
/5Phos/ATCTCTGGTTTTTATCGTGAGGTANNKCTTCAGGGTTACAAGATCGATTGG

P674

ERCAS12A-
885
/5Phos/TCTGGTTTTTATCGTGAGGTAGAANNKCAGGGTTACAAGATCGATTGGACG

P675

ERCAS12A-
886
/5Phos/GGTTTTTATCGTGAGGTAGAACTTNNKGGTTACAAGATCGATTGGACGTAC

P676

ERCAS12A-
887
/5Phos/TTTTATCGTGAGGTAGAACTTCAGNNKTACAAGATCGATTGGACGTACATT

P677

ERCAS12A-
888
/5Phos/TATCGTGAGGTAGAACTTCAGGGTNNKAAGATCGATTGGACGTACATTTCC

P678

ERCAS12A-
889
/5Phos/CGTGAGGTAGAACTTCAGGGTTACNNKATCGATTGGACGTACATTTCCGAG

P679

ERCAS12A-
890
/5Phos/GAGGTAGAACTTCAGGGTTACAAGNNKGATTGGACGTACATTTCCGAGAAA

P680

ERCAS12A-
891
/5Phos/GTAGAACTTCAGGGTTACAAGATCNNKTGGACGTACATTTCCGAGAAAGAC

P681

ERCAS12A-
892
/5Phos/GAACTTCAGGGTTACAAGATCGATNNKACGTACATTTCCGAGAAAGACATT

P682

ERCAS12A-
893
/5Phos/CTTCAGGGTTACAAGATCGATTGGNNKTACATTTCCGAGAAAGACATTGAT

P683

ERCAS12A-
894
/5Phos/CAGGGTTACAAGATCGATTGGACGNNKATTTCCGAGAAAGACATTGATCTT

P684

ERCAS12A-
895
/5Phos/GGTTACAAGATCGATTGGACGTACNNKTCCGAGAAAGACATTGATCTTTTA

P685

ERCAS12A-
896
/5Phos/TACAAGATCGATTGGACGTACATTNNKGAGAAAGACATTGATCTTTTACAA

P686

ERCAS12A-
897
/5Phos/AAGATCGATTGGACGTACATTTCCNNKAAAGACATTGATCTTTTACAAGAG

P687

ERCAS12A-
898
/5Phos/ATCGATTGGACGTACATTTCCGAGNNKGACATTGATCTTTTACAAGAGAAG

P688

ERCAS12A-
899
/5Phos/GATTGGACGTACATTTCCGAGAAANNKATTGATCTTTTACAAGAGAAGGGT

P689

ERCAS12A-
900
/5Phos/TGGACGTACATTTCCGAGAAAGACNNKGATCTTTTACAAGAGAAGGGTCAG

P690

ERCAS12A-
901
/5Phos/ACGTACATTTCCGAGAAAGACATTNNKCTTTTACAAGAGAAGGGTCAGCTT

P691

ERCAS12A-
902
/5Phos/TACATTTCCGAGAAAGACATTGATNNKTTACAAGAGAAGGGTCAGCTTTAC

P692

ERCAS12A-
903
/5Phos/ATTTCCGAGAAAGACATTGATCTTNNKCAAGAGAAGGGTCAGCTTTACTTA

P693

ERCAS12A-
904
/5Phos/TCCGAGAAAGACATTGATCTTTTANNKGAGAAGGGTCAGCTTTACTTATTT

P694

ERCAS12A-
905
/5Phos/GAGAAAGACATTGATCTTTTACAANNKAAGGGTCAGCTTTACTTATTTCAG

P695

ERCAS12A-
906
/5Phos/AAAGACATTGATCTTTTACAAGAGNNKGGTCAGCTTTACTTATTTCAGATC

P696

ERCAS12A-
907
/5Phos/GACATTGATCTTTTACAAGAGAAGNNKCAGCTTTACTTATTTCAGATCTAC

P697

ERCAS12A-
908
/5Phos/ATTGATCTTTTACAAGAGAAGGGTNNKCTTTACTTATTTCAGATCTACAAT

P698

ERCAS12A-
909
/5Phos/GATCTTTTACAAGAGAAGGGTCAGNNKTACTTATTTCAGATCTACAATAAA

P699

ERCAS12A-
910
/5Phos/CTTTTACAAGAGAAGGGTCAGCTTNNKTTATTTCAGATCTACAATAAAGAC

P700

ERCAS12A-
911
/5Phos/TTACAAGAGAAGGGTCAGCTTTACNNKTTTCAGATCTACAATAAAGACTTT

P701

ERCAS12A-
912
/5Phos/CAAGAGAAGGGTCAGCTTTACTTANNKCAGATCTACAATAAAGACTTTTCG

P702

ERCAS12A-
913
/5Phos/GAGAAGGGTCAGCTTTACTTATTTNNKATCTACAATAAAGACTTTTCGAAA

P703

ERCAS12A-
914
/5Phos/AAGGGTCAGCTTTACTTATTTCAGNNKTACAATAAAGACTTTTCGAAAAAA

P704

ERCAS12A-
915
/5Phos/GGTCAGCTTTACTTATTTCAGATCNNKAATAAAGACTTTTCGAAAAAATCC

P705

ERCAS12A-
916
/5Phos/CAGCTTTACTTATTTCAGATCTACNNKAAAGACTTTTCGAAAAAATCCACG

P706

ERCAS12A-
917
/5Phos/CTTTACTTATTTCAGATCTACAATNNKGACTTTTCGAAAAAATCCACGGGC

P707

ERCAS12A-
918
/5Phos/TACTTATTTCAGATCTACAATAAANNKTTTTCGAAAAAATCCACGGGCAAC

P708

ERCAS12A-
919
/5Phos/TTATTTCAGATCTACAATAAAGACNNKTCGAAAAAATCCACGGGCAACGAC

P709

ERCAS12A-
920
/5Phos/TTTCAGATCTACAATAAAGACTTTNNKAAAAAATCCACGGGCAACGACAAC

P710

ERCAS12A-
921
/5Phos/CAGATCTACAATAAAGACTTTTCGNNKAAATCCACGGGCAACGACAACCTT

P711

ERCAS12A-
922
/5Phos/ATCTACAATAAAGACTTTTCGAAANNKTCCACGGGCAACGACAACCTTCAT

P712

ERCAS12A-
923
/5Phos/TACAATAAAGACTTTTCGAAAAAANNKACGGGCAACGACAACCTTCATACA

P713

ERCAS12A-
924
/5Phos/AATAAAGACTTTTCGAAAAAATCCNNKGGCAACGACAACCTTCATACAATG

P714

ERCAS12A-
925
/5Phos/AAAGACTTTTCGAAAAAATCCACGNNKAACGACAACCTTCATACAATGTAC

P715

ERCAS12A-
926
/5Phos/GACTTTTCGAAAAAATCCACGGGCNNKGACAACCTTCATACAATGTACTTA

P716

ERCAS12A-
927
/5Phos/TTTTCGAAAAAATCCACGGGCAACNNKAACCTTCATACAATGTACTTAAAG

P717

ERCAS12A-
928
/5Phos/TCGAAAAAATCCACGGGCAACGACNNKCTTCATACAATGTACTTAAAGAAC

P718

ERCAS12A-
929
/5Phos/AAAAAATCCACGGGCAACGACAACNNKCATACAATGTACTTAAAGAACCTG

P719

ERCAS12A-
930
/5Phos/AAATCCACGGGCAACGACAACCTTNNKACAATGTACTTAAAGAACCTGTTC

P720

ERCAS12A-
931
/5Phos/TCCACGGGCAACGACAACCTTCATNNKATGTACTTAAAGAACCTGTTCTCT

P721

ERCAS12A-
932
/5Phos/ACGGGCAACGACAACCTTCATACANNKTACTTAAAGAACCTGTTCTCTGAA

P722

ERCAS12A-
933
/5Phos/GGCAACGACAACCTTCATACAATGNNKTTAAAGAACCTGTTCTCTGAAGAA

P723

ERCAS12A-
934
/5Phos/AACGACAACCTTCATACAATGTACNNKAAGAACCTGTTCTCTGAAGAAAAC

P724

ERCAS12A-
935
/5Phos/GACAACCTTCATACAATGTACTTANNKAACCTGTTCTCTGAAGAAAACTTG

P725

ERCAS12A-
936
/5Phos/AACCTTCATACAATGTACTTAAAGNNKCTGTTCTCTGAAGAAAACTTGAAA

P726

ERCAS12A-
937
/5Phos/CTTCATACAATGTACTTAAAGAACNNKTTCTCTGAAGAAAACTTGAAAGAT

P727

ERCAS12A-
938
/5Phos/CATACAATGTACTTAAAGAACCTGNNKTCTGAAGAAAACTTGAAAGATATC

P728

ERCAS12A-
939
/5Phos/ACAATGTACTTAAAGAACCTGTTCNNKGAAGAAAACTTGAAAGATATCGTG

P729

ERCAS12A-
940
/5Phos/ATGTACTTAAAGAACCTGTTCTCTNNKGAAAACTTGAAAGATATCGTGCTT

P730

ERCAS12A-
941
/5Phos/TACTTAAAGAACCTGTTCTCTGAANNKAACTTGAAAGATATCGTGCTTAAA

P731

ERCAS12A-
942
/5Phos/TTAAAGAACCTGTTCTCTGAAGAANNKTTGAAAGATATCGTGCTTAAATTG

P732

ERCAS12A-
943
/5Phos/AAGAACCTGTTCTCTGAAGAAAACNNKAAAGATATCGTGCTTAAATTGAAT

P733

ERCAS12A-
944
/5Phos/AACCTGTTCTCTGAAGAAAACTTGNNKGATATCGTGCTTAAATTGAATGGC

P734

ERCAS12A-
945
/5Phos/CTGTTCTCTGAAGAAAACTTGAAANNKATCGTGCTTAAATTGAATGGCGAG

P735

ERCAS12A-
946
/5Phos/TTCTCTGAAGAAAACTTGAAAGATNNKGTGCTTAAATTGAATGGCGAGGCC

P736

ERCAS12A-
947
/5Phos/TCTGAAGAAAACTTGAAAGATATCNNKCTTAAATTGAATGGCGAGGCCGAG

P737

ERCAS12A-
948
/5Phos/GAAGAAAACTTGAAAGATATCGTGNNKAAATTGAATGGCGAGGCCGAGATT

P738

ERCAS12A-
949
/5Phos/GAAAACTTGAAAGATATCGTGCTTNNKTTGAATGGCGAGGCCGAGATTTTT

P739

ERCAS12A-
950
/5Phos/AACTTGAAAGATATCGTGCTTAAANNKAATGGCGAGGCCGAGATTTTTTTC

P740

ERCAS12A-
951
/5Phos/TTGAAAGATATCGTGCTTAAATTGNNKGGCGAGGCCGAGATTTTTTTCCGT

P741

ERCAS12A-
952
/5Phos/AAAGATATCGTGCTTAAATTGAATNNKGAGGCCGAGATTTTTTTCCGTAAA

P742

ERCAS12A-
953
/5Phos/GATATCGTGCTTAAATTGAATGGCNNKGCCGAGATTTTTTTCCGTAAAAGC

P743

ERCAS12A-
954
/5Phos/ATCGTGCTTAAATTGAATGGCGAGNNKGAGATTTTTTTCCGTAAAAGCTCG

P744

ERCAS12A
955
/5Phos/GTGCTTAAATTGAATGGCGAGGCCNNKATTTTTTTCCGTAAAAGCTCGATT

P745

ERCAS12A
956
/5Phos/CTTAAATTGAATGGCGAGGCCGAGNNKTTTTTCCGTAAAAGCTCGATTAAG

P746

ERCAS12A-
957
/5Phos/AAATTGAATGGCGAGGCCGAGATTNNKTTCCGTAAAAGCTCGATTAAGAAT

P747

ERCAS12A-
958
/5Phos/TTGAATGGCGAGGCCGAGATTTTTNNKCGTAAAAGCTCGATTAAGAATCCC

P748

ERCAS12A-
959
/5Phos/AATGGCGAGGCCGAGATTTTTTTCNNKAAAAGCTCGATTAAGAATCCCATC

P749

ERCAS12A-
960
/5Phos/GGCGAGGCCGAGATTTTTTTCCGTNNKAGCTCGATTAAGAATCCCATCATC

P750

ERCAS12A-
961
/5Phos/GAGGCCGAGATTTTTTTCCGTAAANNKTCGATTAAGAATCCCATCATCCAC

P751

ERCAS12A-
962
/5Phos/GCCGAGATTTTTTTCCGTAAAAGCNNKATTAAGAATCCCATCATCCACAAG

P752

ERCAS12A-
963
/5Phos/GAGATTTTTTTCCGTAAAAGCTCGNNKAAGAATCCCATCATCCACAAGAAA

P753

ERCAS12A-
964
/5Phos/ATTTTTTTCCGTAAAAGCTCGATTNNKAATCCCATCATCCACAAGAAAGGC

P754

ERCAS12A-
965
/5Phos/TTTTTCCGTAAAAGCTCGATTAAGNNKCCCATCATCCACAAGAAAGGCAGC

P755

ERCAS12A-
966
/5Phos/TTCCGTAAAAGCTCGATTAAGAATNNKATCATCCACAAGAAAGGCAGCATT

P756

ERCAS12A-
967
/5Phos/CGTAAAAGCTCGATTAAGAATCCCNNKATCCACAAGAAAGGCAGCATTCTG

P757

ERCAS12A-
968
/5Phos/AAAAGCTCGATTAAGAATCCCATCNNKCACAAGAAAGGCAGCATTCTGGTA

P758

ERCAS12A-
969
/5Phos/AGCTCGATTAAGAATCCCATCATCNNKAAGAAAGGCAGCATTCTGGTAAAT

P759

ERCAS12A-
970
/5Phos/TCGATTAAGAATCCCATCATCCACNNKAAAGGCAGCATTCTGGTAAATCGT

P760

ERCAS12A-
971
/5Phos/ATTAAGAATCCCATCATCCACAAGNNKGGCAGCATTCTGGTAAATCGTACC

P761

ERCAS12A-
972
/5Phos/AAGAATCCCATCATCCACAAGAAANNKAGCATTCTGGTAAATCGTACCTAT

P762

ERCAS12A-
973
/5Phos/AATCCCATCATCCACAAGAAAGGCNNKATTCTGGTAAATCGTACCTATGAA

P763

ERCAS12A-
974
/5Phos/CCCATCATCCACAAGAAAGGCAGCNNKCTGGTAAATCGTACCTATGAAGCG

P764

ERCAS12A-
975
/5Phos/ATCATCCACAAGAAAGGCAGCATTNNKGTAAATCGTACCTATGAAGCGGAG

P765

ERCAS12A-
976
/5Phos/ATCCACAAGAAAGGCAGCATTCTGNNKAATCGTACCTATGAAGCGGAGGAA

P766

ERCAS12A-
977
/5Phos/CACAAGAAAGGCAGCATTCTGGTANNKCGTACCTATGAAGCGGAGGAAAAG

P767

ERCAS12A-
978
/5Phos/AAGAAAGGCAGCATTCTGGTAAATNNKACCTATGAAGCGGAGGAAAAGGAC

P768

ERCAS12A-
979
/5Phos/AAAGGCAGCATTCTGGTAAATCGTNNKTATGAAGCGGAGGAAAAGGACCAA

P769

ERCAS12A-
980
/5Phos/GGCAGCATTCTGGTAAATCGTACCNNKGAAGCGGAGGAAAAGGACCAATTT

P770

ERCAS12A-
981
/5Phos/AGCATTCTGGTAAATCGTACCTATNNKGCGGAGGAAAAGGACCAATTTGGT

P771

ERCAS12A-
982
/5Phos/ATTCTGGTAAATCGTACCTATGAANNKGAGGAAAAGGACCAATTTGGTAAT

P772

ERCAS12A-
983
/5Phos/CTGGTAAATCGTACCTATGAAGCGNNKGAAAAGGACCAATTTGGTAATATT

P773

ERCAS12A-
984
/5Phos/GTAAATCGTACCTATGAAGCGGAGNNKAAGGACCAATTTGGTAATATTCAG

P774

ERCAS12A-
985
/5Phos/AATCGTACCTATGAAGCGGAGGAANNKGACCAATTTGGTAATATTCAGATT

P775

ERCAS12A-
986
/5Phos/CGTACCTATGAAGCGGAGGAAAAGNNKCAATTTGGTAATATTCAGATTGTC

P776

ERCAS12A-
987
/5Phos/ACCTATGAAGCGGAGGAAAAGGACNNKTTTGGTAATATTCAGATTGTCCGT

P777

ERCAS12A-
988
/5Phos/TATGAAGCGGAGGAAAAGGACCAANNKGGTAATATTCAGATTGTCCGTAAG

P778

ERCAS12A-
989
/5Phos/GAAGCGGAGGAAAAGGACCAATTTNNKAATATTCAGATTGTCCGTAAGAAC

P779

ERCAS12A-
990
/5Phos/GCGGAGGAAAAGGACCAATTTGGTNNKATTCAGATTGTCCGTAAGAACATC

P780

ERCAS12A-
991
/5Phos/GAGGAAAAGGACCAATTTGGTAATNNKCAGATTGTCCGTAAGAACATCCCT

P781

ERCAS12A-
992
/5Phos/GAAAAGGACCAATTTGGTAATATTNNKATTGTCCGTAAGAACATCCCTGAA

P782

ERCAS12A-
993
/5Phos/AAGGACCAATTTGGTAATATTCAGNNKGTCCGTAAGAACATCCCTGAAAAT

P783

ERCAS12A-
994
/5Phos/GACCAATTTGGTAATATTCAGATTNNKCGTAAGAACATCCCTGAAAATATC

P784

ERCAS12A-
995
/5Phos/CAATTTGGTAATATTCAGATTGTCNNKAAGAACATCCCTGAAAATATCTAT

P785

ERCAS12A-
996
/5Phos/TTTGGTAATATTCAGATTGTCCGTNNKAACATCCCTGAAAATATCTATCAA

P786

ERCAS12A-
997
/5Phos/GGTAATATTCAGATTGTCCGTAAGNNKATCCCTGAAAATATCTATCAAGAA

P787

ERCAS12A-
998
/5Phos/AATATTCAGATTGTCCGTAAGAACNNKCCTGAAAATATCTATCAAGAATTA

P788

ERCAS12A-
999
/5Phos/ATTCAGATTGTCCGTAAGAACATCNNKGAAAATATCTATCAAGAATTATAT

P789

ERCAS12A
1000
/5Phos/CAGATTGTCCGTAAGAACATCCCTNNKAATATCTATCAAGAATTATATAAA

P790

ERCAS12A
1001
/5Phos/ATTGTCCGTAAGAACATCCCTGAANNKATCTATCAAGAATTATATAAATAT

P791

ERCAS12A-
1002
/5Phos/GTCCGTAAGAACATCCCTGAAAATNNKTATCAAGAATTATATAAATATTTC

P792

ERCAS12A-
1003
/5Phos/CGTAAGAACATCCCTGAAAATATCNNKCAAGAATTATATAAATATTTCAAC

P793

ERCAS12A-
1004
/5Phos/AAGAACATCCCTGAAAATATCTATNNKGAATTATATAAATATTTCAACGAT

P794

ERCAS12A-
1005
/5Phos/AACATCCCTGAAAATATCTATCAANNKTTATATAAATATTTCAACGATAAG

P795

ERCAS12A-
1006
/5Phos/ATCCCTGAAAATATCTATCAAGAANNKTATAAATATTTCAACGATAAGTCG

P796

ERCAS12A-
1007
/5Phos/CCTGAAAATATCTATCAAGAATTANNKAAATATTTCAACGATAAGTCGGAC

P797

ERCAS12A-
1008
/5Phos/GAAAATATCTATCAAGAATTATATNNKTATTTCAACGATAAGTCGGACAAG

P798

ERCAS12A-
1009
/5Phos/AATATCTATCAAGAATTATATAAANNKTTCAACGATAAGTCGGACAAGGAA

P799

ERCAS12A-
1010
/5Phos/ATCTATCAAGAATTATATAAATATNNKAACGATAAGTCGGACAAGGAACTT

P800

ERCAS12A-
1011
/5Phos/TATCAAGAATTATATAAATATTTCNNKGATAAGTCGGACAAGGAACTTAGC

P801

ERCAS12A-
1012
/5Phos/CAAGAATTATATAAATATTTCAACNNKAAGTCGGACAAGGAACTTAGCGAT

P802

ERCAS12A-
1013
/5Phos/GAATTATATAAATATTTCAACGATNNKTCGGACAAGGAACTTAGCGATGAA

P803

ERCAS12A-
1014
/5Phos/TTATATAAATATTTCAACGATAAGNNKGACAAGGAACTTAGCGATGAAGCC

P804

ERCAS12A-
1015
/5Phos/TATAAATATTTCAACGATAAGTCGNNKAAGGAACTTAGCGATGAAGCCGCG

P805

ERCAS12A-
1016
/5Phos/AAATATTTCAACGATAAGTCGGACNNKGAACTTAGCGATGAAGCCGCGAAG

P806

ERCAS12A-
1017
/5Phos/TATTTCAACGATAAGTCGGACAAGNNKCTTAGCGATGAAGCCGCGAAGTTG

P807

ERCAS12A-
1018
/5Phos/TTCAACGATAAGTCGGACAAGGAANNKAGCGATGAAGCCGCGAAGTTGAAA

P808

ERCAS12A-
1019
/5Phos/AACGATAAGTCGGACAAGGAACTTNNKGATGAAGCCGCGAAGTTGAAAAAT

P809

ERCAS12A
1020
/5Phos/GATAAGTCGGACAAGGAACTTAGCNNKGAAGCCGCGAAGTTGAAAAATGTT

P810

ERCAS12A-
1021
/5Phos/AAGTCGGACAAGGAACTTAGCGATNNKGCCGCGAAGTTGAAAAATGTTGTT

P811

ERCAS12A-
1022
/5Phos/TCGGACAAGGAACTTAGCGATGAANNKGCGAAGTTGAAAAATGTTGTTGGT

P812

ERCAS12A-
1023
/5Phos/GACAAGGAACTTAGCGATGAAGCCNNKAAGTTGAAAAATGTTGTTGGTCAC

P813

ERCAS12A-
1024
/5Phos/AAGGAACTTAGCGATGAAGCCGCGNNKTTGAAAAATGTTGTTGGTCACCAC

P814

ERCAS12A-
1025
/5Phos/GAACTTAGCGATGAAGCCGCGAAGNNKAAAAATGTTGTTGGTCACCACGAA

P815

ERCAS12A-
1026
/5Phos/CTTAGCGATGAAGCCGCGAAGTTGNNKAATGTTGTTGGTCACCACGAAGCA

P816

ERCAS12A-
1027
/5Phos/AGCGATGAAGCCGCGAAGTTGAAANNKGTTGTTGGTCACCACGAAGCAGCA

P817

ERCAS12A-
1028
/5Phos/GATGAAGCCGCGAAGTTGAAAAATNNKGTTGGTCACCACGAAGCAGCAACA

P818

ERCAS12A-
1029
/5Phos/GAAGCCGCGAAGTTGAAAAATGTTNNKGGTCACCACGAAGCAGCAACAAAC

P819

ERCAS12A-
1030
/5Phos/GCCGCGAAGTTGAAAAATGTTGTTNNKCACCACGAAGCAGCAACAAACATT

P820

ERCAS12A-
1031
/5Phos/GCGAAGTTGAAAAATGTTGTTGGTNNKCACGAAGCAGCAACAAACATTGTG

P821

ERCAS12A-
1032
/5Phos/AAGTTGAAAAATGTTGTTGGTCACNNKGAAGCAGCAACAAACATTGTGAAA

P822

ERCAS12A-
1033
/5Phos/TTGAAAAATGTTGTTGGTCACCACNNKGCAGCAACAAACATTGTGAAAGAC

P823

ERCAS12A-
1034
/5Phos/AAAAATGTTGTTGGTCACCACGAANNKGCAACAAACATTGTGAAAGACTAT

P824

ERCAS12A-
1035
/5Phos/AATGTTGTTGGTCACCACGAAGCANNKACAAACATTGTGAAAGACTATCGC

P825

ERCAS12A-
1036
/5Phos/GTTGTTGGTCACCACGAAGCAGCANNKAACATTGTGAAAGACTATCGCTAC

P826

ERCAS12A-
1037
/5Phos/GTTGGTCACCACGAAGCAGCAACANNKATTGTGAAAGACTATCGCTACACT

P827

ERCAS12A-
1038
/5Phos/GGTCACCACGAAGCAGCAACAAACNNKGTGAAAGACTATCGCTACACTTAC

P828

ERCAS12A-
1039
/5Phos/CACCACGAAGCAGCAACAAACATTNNKAAAGACTATCGCTACACTTACGAT

P829

ERCAS12A-
1040
/5Phos/CACGAAGCAGCAACAAACATTGTGNNKGACTATCGCTACACTTACGATAAG

P830

ERCAS12A-
1041
/5Phos/GAAGCAGCAACAAACATTGTGAAANNKTATCGCTACACTTACGATAAGTAC

P831

ERCAS12A-
1042
/5Phos/GCAGCAACAAACATTGTGAAAGACNNKCGCTACACTTACGATAAGTACTTT

P832

ERCAS12A-
1043
/5Phos/GCAACAAACATTGTGAAAGACTATNNKTACACTTACGATAAGTACTTTTTA

P833

ERCAS12A-
1044
/5Phos/ACAAACATTGTGAAAGACTATCGCNNKACTTACGATAAGTACTTTTTACAT

P834

ERCAS12A-
1045
/5Phos/AACATTGTGAAAGACTATCGCTACNNKTACGATAAGTACTTTTTACATATG

P835

ERCAS12A-
1046
/5Phos/ATTGTGAAAGACTATCGCTACACTNNKGATAAGTACTTTTTACATATGCCT

P836

ERCAS12A-
1047
/5Phos/GTGAAAGACTATCGCTACACTTACNNKAAGTACTTTTTACATATGCCTATT

P837

ERCAS12A-
1048
/5Phos/AAAGACTATCGCTACACTTACGATNNKTACTTTTTACATATGCCTATTACG

P838

ERCAS12A-
1049
/5Phos/GACTATCGCTACACTTACGATAAGNNKTTTTTACATATGCCTATTACGATT

P839

ERCAS12A-
1050
/5Phos/TATCGCTACACTTACGATAAGTACNNKTTACATATGCCTATTACGATTAAC

P840

ERCAS12A-
1051
/5Phos/CGCTACACTTACGATAAGTACTTTNNKCATATGCCTATTACGATTAACTTT

P841

ERCAS12A-
1052
/5Phos/TACACTTACGATAAGTACTTTTTANNKATGCCTATTACGATTAACTTTAAG

P842

ERCAS12A-
1053
/5Phos/ACTTACGATAAGTACTTTTTACATNNKCCTATTACGATTAACTTTAAGGCA

P843

ERCAS12A-
1054
/5Phos/TACGATAAGTACTTTTTACATATGNNKATTACGATTAACTTTAAGGCAAAT

P844

ERCAS12A-
1055
/5Phos/GATAAGTACTTTTTACATATGCCTNNKACGATTAACTTTAAGGCAAATAAG

P845

ERCAS12A-
1056
/5Phos/AAGTACTTTTTACATATGCCTATTNNKATTAACTTTAAGGCAAATAAGACA

P846

ERCAS12A-
1057
/5Phos/TACTTTTTACATATGCCTATTACGNNKAACTTTAAGGCAAATAAGACAGGT

P847

ERCAS12A-
1058
/5Phos/TTTTTACATATGCCTATTACGATTNNKTTTAAGGCAAATAAGACAGGTTTT

P848

ERCAS12A-
1059
/5Phos/TTACATATGCCTATTACGATTAACNNKAAGGCAAATAAGACAGGTTTTATC

P849

ERCAS12A-
1060
/5Phos/CATATGCCTATTACGATTAACTTTNNKGCAAATAAGACAGGTTTTATCAAC

P850

ERCAS12A-
1061
/5Phos/ATGCCTATTACGATTAACTTTAAGNNKAATAAGACAGGTTTTATCAACGAC

P851

ERCAS12A-
1062
/5Phos/CCTATTACGATTAACTTTAAGGCANNKAAGACAGGTTTTATCAACGACCGT

P852

ERCAS12A-
1063
/5Phos/ATTACGATTAACTTTAAGGCAAATNNKACAGGTTTTATCAACGACCGTATC

P853

ERCAS12A-
1064
/5Phos/ACGATTAACTTTAAGGCAAATAAGNNKGGTTTTATCAACGACCGTATCCTG

P854

ERCAS12A-
1065
/5Phos/ATTAACTTTAAGGCAAATAAGACANNKTTTATCAACGACCGTATCCTGCAA

P855

ERCAS12A-
1066
/5Phos/AACTTTAAGGCAAATAAGACAGGTNNKATCAACGACCGTATCCTGCAATAT

P856

ERCAS12A-
1067
/5Phos/TTTAAGGCAAATAAGACAGGTTTTNNKAACGACCGTATCCTGCAATATATT

P857

ERCAS12A-
1068
/5Phos/AAGGCAAATAAGACAGGTTTTATCNNKGACCGTATCCTGCAATATATTGCA

P858

ERCAS12A-
1069
/5Phos/GCAAATAAGACAGGTTTTATCAACNNKCGTATCCTGCAATATATTGCAAAA

P859

ERCAS12A-
1070
/5Phos/AATAAGACAGGTTTTATCAACGACNNKATCCTGCAATATATTGCAAAAGAG

P860

ERCAS12A-
1071
/5Phos/AAGACAGGTTTTATCAACGACCGTNNKCTGCAATATATTGCAAAAGAGAAG

P861

ERCAS12A-
1072
/5Phos/ACAGGTTTTATCAACGACCGTATCNNKCAATATATTGCAAAAGAGAAGGAC

P862

ERCAS12A-
1073
/5Phos/GGTTTTATCAACGACCGTATCCTGNNKTATATTGCAAAAGAGAAGGACCTG

P863

ERCAS12A-
1074
/5Phos/TTTATCAACGACCGTATCCTGCAANNKATTGCAAAAGAGAAGGACCTGCAT

P864

ERCAS12A-
1075
/5Phos/ATCAACGACCGTATCCTGCAATATNNKGCAAAAGAGAAGGACCTGCATGTC

P865

ERCAS12A-
1076
/5Phos/AACGACCGTATCCTGCAATATATTNNKAAAGAGAAGGACCTGCATGTCATT

P866

ERCAS12A-
1077
/5Phos/GACCGTATCCTGCAATATATTGCANNKGAGAAGGACCTGCATGTCATTGGG

P867

ERCAS12A-
1078
/5Phos/CGTATCCTGCAATATATTGCAAAANNKAAGGACCTGCATGTCATTGGGATT

P868

ERCAS12A-
1079
/5Phos/ATCCTGCAATATATTGCAAAAGAGNNKGACCTGCATGTCATTGGGATTGAT

P869

ERCAS12A-
1080
/5Phos/CTGCAATATATTGCAAAAGAGAAGNNKCTGCATGTCATTGGGATTGATCGC

P870

ERCAS12A-
1081
/5Phos/CAATATATTGCAAAAGAGAAGGACNNKCATGTCATTGGGATTGATCGCGGT

P871

ERCAS12A-
1082
/5Phos/TATATTGCAAAAGAGAAGGACCTGNNKGTCATTGGGATTGATCGCGGTGAA

P872

ERCAS12A-
1083
/5Phos/ATTGCAAAAGAGAAGGACCTGCATNNKATTGGGATTGATCGCGGTGAACGC

P873

ERCAS12A-
1084
/5Phos/GCAAAAGAGAAGGACCTGCATGTCNNKGGGATTGATCGCGGTGAACGCAAT

P874

ERCAS12A-
1085
/5Phos/AAAGAGAAGGACCTGCATGTCATTNNKATTGATCGCGGTGAACGCAATCTG

P875

ERCAS12A-
1086
/5Phos/GAGAAGGACCTGCATGTCATTGGGNNKGATCGCGGTGAACGCAATCTGATT

P876

ERCAS12A-
1087
/5Phos/AAGGACCTGCATGTCATTGGGATTNNKCGCGGTGAACGCAATCTGATTTAC

P877

ERCAS12A-
1088
/5Phos/GACCTGCATGTCATTGGGATTGATNNKGGTGAACGCAATCTGATTTACGTG

P878

ERCAS12A-
1089
/5Phos/CTGCATGTCATTGGGATTGATCGCNNKGAACGCAATCTGATTTACGTGAGT

P879

ERCAS12A-
1090
/5Phos/CATGTCATTGGGATTGATCGCGGTNNKCGCAATCTGATTTACGTGAGTGTT

P880

ERCAS12A-
1091
/5Phos/GTCATTGGGATTGATCGCGGTGAANNKAATCTGATTTACGTGAGTGTTATT

P881

ERCAS12A-
1092
/5Phos/ATTGGGATTGATCGCGGTGAACGCNNKCTGATTTACGTGAGTGTTATTGAT

P882

ERCAS12A-
1093
/5Phos/GGGATTGATCGCGGTGAACGCAATNNKATTTACGTGAGTGTTATTGATACC

P883

ERCAS12A-
1094
/5Phos/ATTGATCGCGGTGAACGCAATCTGNNKTACGTGAGTGTTATTGATACCTGC

P884

ERCAS12A-
1095
/5Phos/GATCGCGGTGAACGCAATCTGATTNNKGTGAGTGTTATTGATACCTGCGGT

P885

ERCAS12A-
1096
/5Phos/CGCGGTGAACGCAATCTGATTTACNNKAGTGTTATTGATACCTGCGGTAAT

P886

ERCAS12A-
1097
/5Phos/GGTGAACGCAATCTGATTTACGTGNNKGTTATTGATACCTGCGGTAATATT

P887

ERCAS12A-
1098
/5Phos/GAACGCAATCTGATTTACGTGAGTNNKATTGATACCTGCGGTAATATTGTT

P888

ERCAS12A-
1099
/5Phos/CGCAATCTGATTTACGTGAGTGTTNNKGATACCTGCGGTAATATTGTTGAG

P889

ERCAS12A-
1100
/5Phos/AATCTGATTTACGTGAGTGTTATTNNKACCTGCGGTAATATTGTTGAGCAA

P890

ERCAS12A-
1101
/5Phos/CTGATTTACGTGAGTGTTATTGATNNKTGCGGTAATATTGTTGAGCAAAAG

P891

ERCAS12A-
1102
/5Phos/ATTTACGTGAGTGTTATTGATACCNNKGGTAATATTGTTGAGCAAAAGTCT

P892

ERCAS12A-
1103
/5Phos/TACGTGAGTGTTATTGATACCTGCNNKAATATTGTTGAGCAAAAGTCTTTT

P893

ERCAS12A-
1104
/5Phos/GTGAGTGTTATTGATACCTGCGGTNNKATTGTTGAGCAAAAGTCTTTTAAT

P894

ERCAS12A-
1105
/5Phos/AGTGTTATTGATACCTGCGGTAATNNKGTTGAGCAAAAGTCTTTTAATATC

P895

ERCAS12A-
1106
/5Phos/GTTATTGATACCTGCGGTAATATTNNKGAGCAAAAGTCTTTTAATATCGTT

P896

ERCAS12A-
1107
/5Phos/ATTGATACCTGCGGTAATATTGTTNNKCAAAAGTCTTTTAATATCGTTAAT

P897

ERCAS12A-
1108
/5Phos/GATACCTGCGGTAATATTGTTGAGNNKAAGTCTTTTAATATCGTTAATGGT

P898

ERCAS12A-
1109
/5Phos/ACCTGCGGTAATATTGTTGAGCAANNKTCTTTTAATATCGTTAATGGTTAC

P899

ERCAS12A-
1110
/5Phos/TGCGGTAATATTGTTGAGCAAAAGNNKTTTAATATCGTTAATGGTTACGAC

P900

ERCAS12A-
1111
/5Phos/GGTAATATTGTTGAGCAAAAGTCTNNKAATATCGTTAATGGTTACGACTAC

P901

ERCAS12A-
1112
/5Phos/AATATTGTTGAGCAAAAGTCTTTTNNKATCGTTAATGGTTACGACTACCAA

P902

ERCAS12A-
1113
/5Phos/ATTGTTGAGCAAAAGTCTTTTAATNNKGTTAATGGTTACGACTACCAAATT

P903

ERCAS12A-
1114
/5Phos/GTTGAGCAAAAGTCTTTTAATATCNNKAATGGTTACGACTACCAAATTAAG

P904

ERCAS12A-
1115
/5Phos/GAGCAAAAGTCTTTTAATATCGTTNNKGGTTACGACTACCAAATTAAGCTT

P905

ERCAS12A-
1116
/5Phos/CAAAAGTCTTTTAATATCGTTAATNNKTACGACTACCAAATTAAGCTTAAA

P906

ERCAS12A-
1117
/5Phos/AAGTCTTTTAATATCGTTAATGGTNNKGACTACCAAATTAAGCTTAAACAA

P907

ERCAS12A-
1118
/5Phos/TCTTTTAATATCGTTAATGGTTACNNKTACCAAATTAAGCTTAAACAACAG

P908

ERCAS12A-
1119
/5Phos/TTTAATATCGTTAATGGTTACGACNNKCAAATTAAGCTTAAACAACAGGAA

P909

ERCAS12A-
1120
/5Phos/AATATCGTTAATGGTTACGACTACNNKATTAAGCTTAAACAACAGGAAGGT

P910

ERCAS12A-
1121
/5Phos/ATCGTTAATGGTTACGACTACCAANNKAAGCTTAAACAACAGGAAGGTGCC

P911

ERCAS12A-
1122
/5Phos/GTTAATGGTTACGACTACCAAATTNNKCTTAAACAACAGGAAGGTGCCCGC

P912

ERCAS12A-
1123
/5Phos/AATGGTTACGACTACCAAATTAAGNNKAAACAACAGGAAGGTGCCCGCCAG

P913

ERCAS12A-
1124
/5Phos/GGTTACGACTACCAAATTAAGCTTNNKCAACAGGAAGGTGCCCGCCAGATC

P914

ERCAS12A-
1125
/5Phos/TACGACTACCAAATTAAGCTTAAANNKCAGGAAGGTGCCCGCCAGATCGCC

P915

ERCAS12A-
1126
/5Phos/GACTACCAAATTAAGCTTAAACAANNKGAAGGTGCCCGCCAGATCGCCCGT

P916

ERCAS12A-
1127
/5Phos/TACCAAATTAAGCTTAAACAACAGNNKGGTGCCCGCCAGATCGCCCGTAAG

P917

ERCAS12A
1128
/5Phos/CAAATTAAGCTTAAACAACAGGAANNKGCCCGCCAGATCGCCCGTAAGGAG

P918

ERCAS12A-
1129
/5Phos/ATTAAGCTTAAACAACAGGAAGGTNNKCGCCAGATCGCCCGTAAGGAGTGG

P919

ERCAS12A-
1130
/5Phos/AAGCTTAAACAACAGGAAGGTGCCNNKCAGATCGCCCGTAAGGAGTGGAAG

P920

ERCAS12A-
1131
/5Phos/CTTAAACAACAGGAAGGTGCCCGCNNKATCGCCCGTAAGGAGTGGAAGGAA

P921

ERCAS12A-
1132
/5Phos/AAACAACAGGAAGGTGCCCGCCAGNNKGCCCGTAAGGAGTGGAAGGAAATC

P922

ERCAS12A-
1133
/5Phos/CAACAGGAAGGTGCCCGCCAGATCNNKCGTAAGGAGTGGAAGGAAATCGGA

P923

ERCAS12A-
1134
/5Phos/CAGGAAGGTGCCCGCCAGATCGCCNNKAAGGAGTGGAAGGAAATCGGAAAA

P924

ERCAS12A-
1135
/5Phos/GAAGGTGCCCGCCAGATCGCCCGTNNKGAGTGGAAGGAAATCGGAAAAATT

P925

ERCAS12A-
1136
/5Phos/GGTGCCCGCCAGATCGCCCGTAAGNNKTGGAAGGAAATCGGAAAAATTAAG

P926

ERCAS12A-
1137
/5Phos/GCCCGCCAGATCGCCCGTAAGGAGNNKAAGGAAATCGGAAAAATTAAGGAA

P927

ERCAS12A-
1138
/5Phos/CGCCAGATCGCCCGTAAGGAGTGGNNKGAAATCGGAAAAATTAAGGAAATC

P928

ERCAS12A-
1139
/5Phos/CAGATCGCCCGTAAGGAGTGGAAGNNKATCGGAAAAATTAAGGAAATCAAA

P929

ERCAS12A-
1140
/5Phos/ATCGCCCGTAAGGAGTGGAAGGAANNKGGAAAAATTAAGGAAATCAAAGAG

P930

ERCAS12A-
1141
/5Phos/GCCCGTAAGGAGTGGAAGGAAATCNNKAAAATTAAGGAAATCAAAGAGGGA

P931

ERCAS12A-
1142
/5Phos/CGTAAGGAGTGGAAGGAAATCGGANNKATTAAGGAAATCAAAGAGGGATAT

P932

ERCAS12A-
1143
/5Phos/AAGGAGTGGAAGGAAATCGGAAAANNKAAGGAAATCAAAGAGGGATATCTG

P933

ERCAS12A-
1144
/5Phos/GAGTGGAAGGAAATCGGAAAAATTNNKGAAATCAAAGAGGGATATCTGAGC

P934

ERCAS12A-
1145
/5Phos/TGGAAGGAAATCGGAAAAATTAAGNNKATCAAAGAGGGATATCTGAGCTTG

P935

ERCAS12A-
1146
/5Phos/AAGGAAATCGGAAAAATTAAGGAANNKAAAGAGGGATATCTGAGCTTGGTT

P936

ERCAS12A-
1147
/5Phos/GAAATCGGAAAAATTAAGGAAATCNNKGAGGGATATCTGAGCTTGGTTATT

P937

ERCAS12A-
1148
/5Phos/ATCGGAAAAATTAAGGAAATCAAANNKGGATATCTGAGCTTGGTTATTCAT

P938

ERCAS12A-
1149
/5Phos/GGAAAAATTAAGGAAATCAAAGAGNNKTATCTGAGCTTGGTTATTCATGAA

P939

ERCAS12A-
1150
/5Phos/AAAATTAAGGAAATCAAAGAGGGANNKCTGAGCTTGGTTATTCATGAAATC

P940

ERCAS12A-
1151
/5Phos/ATTAAGGAAATCAAAGAGGGATATNNKAGCTTGGTTATTCATGAAATCTCA

P941

ERCAS12A-
1152
/5Phos/AAGGAAATCAAAGAGGGATATCTGNNKTTGGTTATTCATGAAATCTCAAAG

P942

ERCAS12A-
1153
/5Phos/GAAATCAAAGAGGGATATCTGAGCNNKGTTATTCATGAAATCTCAAAGATG

P943

ERCAS12A-
1154
/5Phos/ATCAAAGAGGGATATCTGAGCTTGNNKATTCATGAAATCTCAAAGATGGTT

P944

ERCAS12A-
1155
/5Phos/AAAGAGGGATATCTGAGCTTGGTTNNKCATGAAATCTCAAAGATGGTTATT

P945

ERCAS12A-
1156
/5Phos/GAGGGATATCTGAGCTTGGTTATTNNKGAAATCTCAAAGATGGTTATTAAG

P946

ERCAS12A-
1157
/5Phos/GGATATCTGAGCTTGGTTATTCATNNKATCTCAAAGATGGTTATTAAGTAC

P947

ERCAS12A-
1158
/5Phos/TATCTGAGCTTGGTTATTCATGAANNKTCAAAGATGGTTATTAAGTACAAC

P948

ERCAS12A-
1159
/5Phos/CTGAGCTTGGTTATTCATGAAATCNNKAAGATGGTTATTAAGTACAACGCC

P949

ERCAS12A-
1160
/5Phos/AGCTTGGTTATTCATGAAATCTCANNKATGGTTATTAAGTACAACGCCATC

P950

ERCAS12A-
1161
/5Phos/TTGGTTATTCATGAAATCTCAAAGNNKGTTATTAAGTACAACGCCATCATC

P951

ERCAS12A-
1162
/5Phos/GTTATTCATGAAATCTCAAAGATGNNKATTAAGTACAACGCCATCATCGCT

P952

ERCAS12A-
1163
/5Phos/ATTCATGAAATCTCAAAGATGGTTNNKAAGTACAACGCCATCATCGCTATG

P953

ERCAS12A-
1164
/5Phos/CATGAAATCTCAAAGATGGTTATTNNKTACAACGCCATCATCGCTATGGAA

P954

ERCAS12A-
1165
/5Phos/GAAATCTCAAAGATGGTTATTAAGNNKAACGCCATCATCGCTATGGAAGAC

P955

ERCAS12A-
1166
/5Phos/ATCTCAAAGATGGTTATTAAGTACNNKGCCATCATCGCTATGGAAGACTTA

P956

ERCAS12A-
1167
/5Phos/TCAAAGATGGTTATTAAGTACAACNNKATCATCGCTATGGAAGACTTATCG

P957

ERCAS12A
1168
/5Phos/AAGATGGTTATTAAGTACAACGCCNNKATCGCTATGGAAGACTTATCGTAT

P958

ERCAS12A-
1169
/5Phos/ATGGTTATTAAGTACAACGCCATCNNKGCTATGGAAGACTTATCGTATGGG

P959

ERCAS12A-
1170
/5Phos/GTTATTAAGTACAACGCCATCATCNNKATGGAAGACTTATCGTATGGGTTT

P960

ERCAS12A-
1171
/5Phos/ATTAAGTACAACGCCATCATCGCTNNKGAAGACTTATCGTATGGGTTTAAA

P961

ERCAS12A
1172
/5Phos/AAGTACAACGCCATCATCGCTATGNNKGACTTATCGTATGGGTTTAAAAAG

P962

ERCAS12A-
1173
/5Phos/TACAACGCCATCATCGCTATGGAANNKTTATCGTATGGGTTTAAAAAGGGT

P963

ERCAS12A-
1174
/5Phos/AACGCCATCATCGCTATGGAAGACNNKTCGTATGGGTTTAAAAAGGGTCGT

P964

ERCAS12A-
1175
/5Phos/GCCATCATCGCTATGGAAGACTTANNKTATGGGTTTAAAAAGGGTCGTTTC

P965

ERCAS12A-
1176
/5Phos/ATCATCGCTATGGAAGACTTATCGNNKGGGTTTAAAAAGGGTCGTTTCAAA

P966

ERCAS12A-
1177
/5Phos/ATCGCTATGGAAGACTTATCGTATNNKTTTAAAAAGGGTCGTTTCAAAGTT

P967

ERCAS12A-
1178
/5Phos/GCTATGGAAGACTTATCGTATGGGNNKAAAAAGGGTCGTTTCAAAGTTGAG

P968

ERCAS12A-
1179
/5Phos/ATGGAAGACTTATCGTATGGGTTTNNKAAGGGTCGTTTCAAAGTTGAGCGC

P969

ERCAS12A-
1180
/5Phos/GAAGACTTATCGTATGGGTTTAAANNKGGTCGTTTCAAAGTTGAGCGCCAG

P970

ERCAS12A-
1181
/5Phos/GACTTATCGTATGGGTTTAAAAAGNNKCGTTTCAAAGTTGAGCGCCAGGTC

P971

ERCAS12A-
1182
/5Phos/TTATCGTATGGGTTTAAAAAGGGTNNKTTCAAAGTTGAGCGCCAGGTCTAC

P972

ERCAS12A-
1183
/5Phos/TCGTATGGGTTTAAAAAGGGTCGTNNKAAAGTTGAGCGCCAGGTCTACCAA

P973

ERCAS12A-
1184
/5Phos/TATGGGTTTAAAAAGGGTCGTTTCNNKGTTGAGCGCCAGGTCTACCAAAAG

P974

ERCAS12A-
1185
/5Phos/GGGTTTAAAAAGGGTCGTTTCAAANNKGAGCGCCAGGTCTACCAAAAGTTT

P975

ERCAS12A-
1186
/5Phos/TTTAAAAAGGGTCGTTTCAAAGTTNNKCGCCAGGTCTACCAAAAGTTTGAA

P976

ERCAS12A-
1187
/5Phos/AAAAAGGGTCGTTTCAAAGTTGAGNNKCAGGTCTACCAAAAGTTTGAAACG

P977

ERCAS12A-
1188
/5Phos/AAGGGTCGTTTCAAAGTTGAGCGCNNKGTCTACCAAAAGTTTGAAACGATG

P978

ERCAS12A-
1189
/5Phos/GGTCGTTTCAAAGTTGAGCGCCAGNNKTACCAAAAGTTTGAAACGATGCTT

P979

ERCAS12A-
1190
/5Phos/CGTTTCAAAGTTGAGCGCCAGGTCNNKCAAAAGTTTGAAACGATGCTTATT

P980

ERCAS12A-
1191
/5Phos/TTCAAAGTTGAGCGCCAGGTCTACNNKAAGTTTGAAACGATGCTTATTAAT

P981

ERCAS12A-
1192
/5Phos/AAAGTTGAGCGCCAGGTCTACCAANNKTTTGAAACGATGCTTATTAATAAA

P982

ERCAS12A-
1193
/5Phos/GTTGAGCGCCAGGTCTACCAAAAGNNKGAAACGATGCTTATTAATAAATTA

P983

ERCAS12A-
1194
/5Phos/GAGCGCCAGGTCTACCAAAAGTTTNNKACGATGCTTATTAATAAATTAAAC

P984

ERCAS12A-
1195
/5Phos/CGCCAGGTCTACCAAAAGTTTGAANNKATGCTTATTAATAAATTAAACTAC

P985

ERCAS12A-
1196
/5Phos/CAGGTCTACCAAAAGTTTGAAACGNNKCTTATTAATAAATTAAACTACTTG

P986

ERCAS12A-
1197
/5Phos/GTCTACCAAAAGTTTGAAACGATGNNKATTAATAAATTAAACTACTTGGTT

P987

ERCAS12A-
1198
/5Phos/TACCAAAAGTTTGAAACGATGCTTNNKAATAAATTAAACTACTTGGTTTTC

P988

ERCAS12A-
1199
/5Phos/CAAAAGTTTGAAACGATGCTTATTNNKAAATTAAACTACTTGGTTTTCAAA

P989

ERCAS12A-
1200
/5Phos/AAGTTTGAAACGATGCTTATTAATNNKTTAAACTACTTGGTTTTCAAAGAT

P990

ERCAS12A-
1201
/5Phos/TTTGAAACGATGCTTATTAATAAANNKAACTACTTGGTTTTCAAAGATATC

P991

ERCAS12A-
1202
/5Phos/GAAACGATGCTTATTAATAAATTANNKTACTTGGTTTTCAAAGATATCTCT

P992

ERCAS12A-
1203
/5Phos/ACGATGCTTATTAATAAATTAAACNNKTTGGTTTTCAAAGATATCTCTATT

P993

ERCAS12A-
1204
/5Phos/ATGCTTATTAATAAATTAAACTACNNKGTTTTCAAAGATATCTCTATTACC

P994

ERCAS12A-
1205
/5Phos/CTTATTAATAAATTAAACTACTTGNNKTTCAAAGATATCTCTATTACCGAG

P995

ERCAS12A-
1206
/5Phos/ATTAATAAATTAAACTACTTGGTTNNKAAAGATATCTCTATTACCGAGAAT

P996

ERCAS12A-
1207
/5Phos/AATAAATTAAACTACTTGGTTTTCNNKGATATCTCTATTACCGAGAATGGA

P997

ERCAS12A-
1208
/5Phos/AAATTAAACTACTTGGTTTTCAAANNKATCTCTATTACCGAGAATGGAGGT

P998

ERCAS12A-
1209
/5Phos/TTAAACTACTTGGTTTTCAAAGATNNKTCTATTACCGAGAATGGAGGTTTA

P999

ERCAS12A-
1210
/5Phos/AACTACTTGGTTTTCAAAGATATCNNKATTACCGAGAATGGAGGTTTACTG

P1000

ERCAS12A-
1211
/5Phos/TACTTGGTTTTCAAAGATATCTCTNNKACCGAGAATGGAGGTTTACTGAAA

P1001

ERCAS12A-
1212
/5Phos/TTGGTTTTCAAAGATATCTCTATTNNKGAGAATGGAGGTTTACTGAAAGGA

P1002

ERCAS12A-
1213
/5Phos/GTTTTCAAAGATATCTCTATTACCNNKAATGGAGGTTTACTGAAAGGATAC

P1003

ERCAS12A-
1214
/5Phos/TTCAAAGATATCTCTATTACCGAGNNKGGAGGTTTACTGAAAGGATACCAG

P1004

ERCAS12A-
1215
/5Phos/AAAGATATCTCTATTACCGAGAATNNKGGTTTACTGAAAGGATACCAGTTA

P1005

ERCAS12A-
1216
/5Phos/GATATCTCTATTACCGAGAATGGANNKTTACTGAAAGGATACCAGTTAACG

P1006

ERCAS12A-
1217
/5Phos/ATCTCTATTACCGAGAATGGAGGTNNKCTGAAAGGATACCAGTTAACGTAT

P1007

ERCAS12A-
1218
/5Phos/TCTATTACCGAGAATGGAGGTTTANNKAAAGGATACCAGTTAACGTATATC

P1008

ERCAS12A-
1219
/5Phos/ATTACCGAGAATGGAGGTTTACTGNNKGGATACCAGTTAACGTATATCCCC

P1009

ERCAS12A-
1220
/5Phos/ACCGAGAATGGAGGTTTACTGAAANNKTACCAGTTAACGTATATCCCCGAT

P1010

ERCAS12A-
1221
/5Phos/GAGAATGGAGGTTTACTGAAAGGANNKCAGTTAACGTATATCCCCGATAAG

P1011

ERCAS12A-
1222
/5Phos/AATGGAGGTTTACTGAAAGGATACNNKTTAACGTATATCCCCGATAAGCTT

P1012

ERCAS12A-
1223
/5Phos/GGAGGTTTACTGAAAGGATACCAGNNKACGTATATCCCCGATAAGCTTAAA

P1013

ERCAS12A-
1224
/5Phos/GGTTTACTGAAAGGATACCAGTTANNKTATATCCCCGATAAGCTTAAAAAC

P1014

ERCAS12A-
1225
/5Phos/TTACTGAAAGGATACCAGTTAACGNNKATCCCCGATAAGCTTAAAAACGTG

P1015

ERCAS12A-
1226
/5Phos/CTGAAAGGATACCAGTTAACGTATNNKCCCGATAAGCTTAAAAACGTGGGC

P1016

ERCAS12A-
1227
/5Phos/AAAGGATACCAGTTAACGTATATCNNKGATAAGCTTAAAAACGTGGGCCAT

P1017

ERCAS12A-
1228
/5Phos/GGATACCAGTTAACGTATATCCCCNNKAAGCTTAAAAACGTGGGCCATCAG

P1018

ERCAS12A-
1229
/5Phos/TACCAGTTAACGTATATCCCCGATNNKCTTAAAAACGTGGGCCATCAGTGT

P1019

ERCAS12A-
1230
/5Phos/CAGTTAACGTATATCCCCGATAAGNNKAAAAACGTGGGCCATCAGTGTGGC

P1020

ERCAS12A-
1231
/5Phos/TTAACGTATATCCCCGATAAGCTTNNKAACGTGGGCCATCAGTGTGGCTGT

P1021

ERCAS12A-
1232
/5Phos/ACGTATATCCCCGATAAGCTTAAANNKGTGGGCCATCAGTGTGGCTGTATT

P1022

ERCAS12A-
1233
/5Phos/TATATCCCCGATAAGCTTAAAAACNNKGGCCATCAGTGTGGCTGTATTTTC

P1023

ERCAS12A-
1234
/5Phos/ATCCCCGATAAGCTTAAAAACGTGNNKCATCAGTGTGGCTGTATTTTCTAT

P1024

ERCAS12A-
1235
/5Phos/CCCGATAAGCTTAAAAACGTGGGCNNKCAGTGTGGCTGTATTTTCTATGTG

P1025

ERCAS12A-
1236
/5Phos/GATAAGCTTAAAAACGTGGGCCATNNKTGTGGCTGTATTTTCTATGTGCCC

P1026

ERCAS12A-
1237
/5Phos/AAGCTTAAAAACGTGGGCCATCAGNNKGGCTGTATTTTCTATGTGCCCGCG

P1027

ERCAS12A-
1238
/5Phos/CTTAAAAACGTGGGCCATCAGTGTNNKTGTATTTTCTATGTGCCCGCGGCA

P1028

ERCAS12A-
1239
/5Phos/AAAAACGTGGGCCATCAGTGTGGCNNKATTTTCTATGTGCCCGCGGCATAT

P1029

ERCAS12A-
1240
/5Phos/AACGTGGGCCATCAGTGTGGCTGTNNKTTCTATGTGCCCGCGGCATATACA

P1030

ERCAS12A-
1241
/5Phos/GTGGGCCATCAGTGTGGCTGTATTNNKTATGTGCCCGCGGCATATACATCA

P1031

ERCAS12A-
1242
/5Phos/GGCCATCAGTGTGGCTGTATTTTCNNKGTGCCCGCGGCATATACATCAAAA

P1032

ERCAS12A-
1243
/5Phos/CATCAGTGTGGCTGTATTTTCTATNNKCCCGCGGCATATACATCAAAAATT

P1033

ERCAS12A-
1244
/5Phos/CAGTGTGGCTGTATTTTCTATGTGNNKGCGGCATATACATCAAAAATTGAC

P1034

ERCAS12A-
1245
/5Phos/TGTGGCTGTATTTTCTATGTGCCCNNKGCATATACATCAAAAATTGACCCG

P1035

ERCAS12A-
1246
/5Phos/GGCTGTATTTTCTATGTGCCCGCGNNKTATACATCAAAAATTGACCCGACG

P1036

ERCAS12A-
1247
/5Phos/TGTATTTTCTATGTGCCCGCGGCANNKACATCAAAAATTGACCCGACGACG

P1037

ERCAS12A-
1248
/5Phos/ATTTTCTATGTGCCCGCGGCATATNNKTCAAAAATTGACCCGACGACGGGT

P1038

ERCAS12A-
1249
/5Phos/TTCTATGTGCCCGCGGCATATACANNKAAAATTGACCCGACGACGGGTTTC

P1039

ERCAS12A-
1250
/5Phos/TATGTGCCCGCGGCATATACATCANNKATTGACCCGACGACGGGTTTCGTT

P1040

ERCAS12A-
1251
/5Phos/GTGCCCGCGGCATATACATCAAAANNKGACCCGACGACGGGTTTCGTTAAT

P1041

ERCAS12A-
1252
/5Phos/CCCGCGGCATATACATCAAAAATTNNKCCGACGACGGGTTTCGTTAATATC

P1042

ERCAS12A-
1253
/5Phos/GCGGCATATACATCAAAAATTGACNNKACGACGGGTTTCGTTAATATCTTC

P1043

ERCAS12A-
1254
/5Phos/GCATATACATCAAAAATTGACCCGNNKACGGGTTTCGTTAATATCTTCAAG

P1044

ERCAS12A-
1255
/5Phos/TATACATCAAAAATTGACCCGACGNNKGGTTTCGTTAATATCTTCAAGTTC

P1045

ERCAS12A-
1256
/5Phos/ACATCAAAAATTGACCCGACGACGNNKTTCGTTAATATCTTCAAGTTCAAA

P1046

ERCAS12A-
1257
/5Phos/TCAAAAATTGACCCGACGACGGGTNNKGTTAATATCTTCAAGTTCAAAGAT

P1047

ERCAS12A-
1258
/5Phos/AAAATTGACCCGACGACGGGTTTCNNKAATATCTTCAAGTTCAAAGATCTG

P1048

ERCAS12A-
1259
/5Phos/ATTGACCCGACGACGGGTTTCGTTNNKATCTTCAAGTTCAAAGATCTGACA

P1049

ERCAS12A-
1260
/5Phos/GACCCGACGACGGGTTTCGTTAATNNKTTCAAGTTCAAAGATCTGACAGTT

P1050

ERCAS12A-
1261
/5Phos/CCGACGACGGGTTTCGTTAATATCNNKAAGTTCAAAGATCTGACAGTTGAT

P1051

ERCAS12A-
1262
/5Phos/ACGACGGGTTTCGTTAATATCTTCNNKTTCAAAGATCTGACAGTTGATGCA

P1052

ERCAS12A-
1263
/5Phos/ACGGGTTTCGTTAATATCTTCAAGNNKAAAGATCTGACAGTTGATGCAAAA

P1053

ERCAS12A-
1264
/5Phos/GGTTTCGTTAATATCTTCAAGTTCNNKGATCTGACAGTTGATGCAAAACGC

P1054

ERCAS12A-
1265
/5Phos/TTCGTTAATATCTTCAAGTTCAAANNKCTGACAGTTGATGCAAAACGCGAA

P1055

ERCAS12A-
1266
/5Phos/GTTAATATCTTCAAGTTCAAAGATNNKACAGTTGATGCAAAACGCGAATTC

P1056

ERCAS12A-
1267
/5Phos/AATATCTTCAAGTTCAAAGATCTGNNKGTTGATGCAAAACGCGAATTCATT

P1057

ERCAS12A-
1268
/5Phos/ATCTTCAAGTTCAAAGATCTGACANNKGATGCAAAACGCGAATTCATTAAG

P1058

ERCAS12A-
1269
/5Phos/TTCAAGTTCAAAGATCTGACAGTTNNKGCAAAACGCGAATTCATTAAGAAG

P1059

ERCAS12A-
1270
/5Phos/AAGTTCAAAGATCTGACAGTTGATNNKAAACGCGAATTCATTAAGAAGTTC

P1060

ERCAS12A-
1271
/5Phos/TTCAAAGATCTGACAGTTGATGCANNKCGCGAATTCATTAAGAAGTTCGAT

P1061

ERCAS12A-
1272
/5Phos/AAAGATCTGACAGTTGATGCAAAANNKGAATTCATTAAGAAGTTCGATAGT

P1062

ERCAS12A-
1273
/5Phos/GATCTGACAGTTGATGCAAAACGCNNKTTCATTAAGAAGTTCGATAGTATT

P1063

ERCAS12A-
1274
/5Phos/CTGACAGTTGATGCAAAACGCGAANNKATTAAGAAGTTCGATAGTATTCGC

P1064

ERCAS12A-
1275
/5Phos/ACAGTTGATGCAAAACGCGAATTCNNKAAGAAGTTCGATAGTATTCGCTAT

P1065

ERCAS12A-
1276
/5Phos/GTTGATGCAAAACGCGAATTCATTNNKAAGTTCGATAGTATTCGCTATGAC

P1066

ERCAS12A-
1277
/5Phos/GATGCAAAACGCGAATTCATTAAGNNKTTCGATAGTATTCGCTATGACTCC

P1067

ERCAS12A-
1278
/5Phos/GCAAAACGCGAATTCATTAAGAAGNNKGATAGTATTCGCTATGACTCCGAG

P1068

ERCAS12A-
1279
/5Phos/AAACGCGAATTCATTAAGAAGTTCNNKAGTATTCGCTATGACTCCGAGAAA

P1069

ERCAS12A-
1280
/5Phos/CGCGAATTCATTAAGAAGTTCGATNNKATTCGCTATGACTCCGAGAAAAAT

P1070

ERCAS12A-
1281
/5Phos/GAATTCATTAAGAAGTTCGATAGTNNKCGCTATGACTCCGAGAAAAATTTG

P1071

ERCAS12A-
1282
/5Phos/TTCATTAAGAAGTTCGATAGTATTNNKTATGACTCCGAGAAAAATTTGTTT

P1072

ERCAS12A-
1283
/5Phos/ATTAAGAAGTTCGATAGTATTCGCNNKGACTCCGAGAAAAATTTGTTTTGC

P1073

ERCAS12A-
1284
/5Phos/AAGAAGTTCGATAGTATTCGCTATNNKTCCGAGAAAAATTTGTTTTGCTTC

P1074

ERCAS12A-
1285
/5Phos/AAGTTCGATAGTATTCGCTATGACNNKGAGAAAAATTTGTTTTGCTTCACT

P1075

ERCAS12A-
1286
/5Phos/TTCGATAGTATTCGCTATGACTCCNNKAAAAATTTGTTTTGCTTCACTTTT

P1076

ERCAS12A-
1287
/5Phos/GATAGTATTCGCTATGACTCCGAGNNKAATTTGTTTTGCTTCACTTTTGAC

P1077

ERCAS12A-
1288
/5Phos/AGTATTCGCTATGACTCCGAGAAANNKTTGTTTTGCTTCACTTTTGACTAC

P1078

ERCAS12A-
1289
/5Phos/ATTCGCTATGACTCCGAGAAAAATNNKTTTTGCTTCACTTTTGACTACAAT

P1079

ERCAS12A-
1290
/5Phos/CGCTATGACTCCGAGAAAAATTTGNNKTGCTTCACTTTTGACTACAATAAC

P1080

ERCAS12A-
1291
/5Phos/TATGACTCCGAGAAAAATTTGTTTNNKTTCACTTTTGACTACAATAACTTT

P1081

ERCAS12A-
1292
/5Phos/GACTCCGAGAAAAATTTGTTTTGCNNKACTTTTGACTACAATAACTTTATT

P1082

ERCAS12A-
1293
/5Phos/TCCGAGAAAAATTTGTTTTGCTTCNNKTTTGACTACAATAACTTTATTACT

P1083

ERCAS12A-
1294
/5Phos/GAGAAAAATTTGTTTTGCTTCACTNNKGACTACAATAACTTTATTACTCAG

P1084

ERCAS12A-
1295
/5Phos/AAAAATTTGTTTTGCTTCACTTTTNNKTACAATAACTTTATTACTCAGAAT

P1085

ERCAS12A-
1296
/5Phos/AATTTGTTTTGCTTCACTTTTGACNNKAATAACTTTATTACTCAGAATACA

P1086

ERCAS12A-
1297
/5Phos/TTGTTTTGCTTCACTTTTGACTACNNKAACTTTATTACTCAGAATACAGTC

P1087

ERCAS12A-
1298
/5Phos/TTTTGCTTCACTTTTGACTACAATNNKTTTATTACTCAGAATACAGTCATG

P1088

ERCAS12A-
1299
/5Phos/TGCTTCACTTTTGACTACAATAACNNKATTACTCAGAATACAGTCATGAGT

P1089

ERCAS12A-
1300
/5Phos/TTCACTTTTGACTACAATAACTTTNNKACTCAGAATACAGTCATGAGTAAA

P1090

ERCAS12A-
1301
/5Phos/ACTTTTGACTACAATAACTTTATTNNKCAGAATACAGTCATGAGTAAAAGC

P1091

ERCAS12A-
1302
/5Phos/TTTGACTACAATAACTTTATTACTNNKAATACAGTCATGAGTAAAAGCTCC

P1092

ERCAS12A-
1303
/5Phos/GACTACAATAACTTTATTACTCAGNNKACAGTCATGAGTAAAAGCTCCTGG

P1093

ERCAS12A-
1304
/5Phos/TACAATAACTTTATTACTCAGAATNNKGTCATGAGTAAAAGCTCCTGGTCG

P1094

ERCAS12A-
1305
/5Phos/AATAACTTTATTACTCAGAATACANNKATGAGTAAAAGCTCCTGGTCGGTT

P1095

ERCAS12A-
1306
/5Phos/AACTTTATTACTCAGAATACAGTCNNKAGTAAAAGCTCCTGGTCGGTTTAC

P1096

ERCAS12A-
1307
/5Phos/TTTATTACTCAGAATACAGTCATGNNKAAAAGCTCCTGGTCGGTTTACACG

P1097

ERCAS12A-
1308
/5Phos/ATTACTCAGAATACAGTCATGAGTNNKAGCTCCTGGTCGGTTTACACGTAC

P1098

ERCAS12A-
1309
/5Phos/ACTCAGAATACAGTCATGAGTAAANNKTCCTGGTCGGTTTACACGTACGGG

P1099

ERCAS12A-
1310
/5Phos/CAGAATACAGTCATGAGTAAAAGCNNKTGGTCGGTTTACACGTACGGGGTA

P1100

ERCAS12A-
1311
/5Phos/AATACAGTCATGAGTAAAAGCTCCNNKTCGGTTTACACGTACGGGGTACGC

P1101

ERCAS12A-
1312
/5Phos/ACAGTCATGAGTAAAAGCTCCTGGNNKGTTTACACGTACGGGGTACGCATC

P1102

ERCAS12A-
1313
/5Phos/GTCATGAGTAAAAGCTCCTGGTCGNNKTACACGTACGGGGTACGCATCAAA

P1103

ERCAS12A-
1314
/5Phos/ATGAGTAAAAGCTCCTGGTCGGTTNNKACGTACGGGGTACGCATCAAACGT

P1104

ERCAS12A-
1315
/5Phos/AGTAAAAGCTCCTGGTCGGTTTACNNKTACGGGGTACGCATCAAACGTCGT

P1105

ERCAS12A-
1316
/5Phos/AAAAGCTCCTGGTCGGTTTACACGNNKGGGGTACGCATCAAACGTCGTTTT

P1106

ERCAS12A-
1317
/5Phos/AGCTCCTGGTCGGTTTACACGTACNNKGTACGCATCAAACGTCGTTTTGTG

P1107

ERCAS12A-
1318
/5Phos/TCCTGGTCGGTTTACACGTACGGGNNKCGCATCAAACGTCGTTTTGTGAAT

P1108

ERCAS12A-
1319
/5Phos/TGGTCGGTTTACACGTACGGGGTANNKATCAAACGTCGTTTTGTGAATGGT

P1109

ERCAS12A-
1320
/5Phos/TCGGTTTACACGTACGGGGTACGCNNKAAACGTCGTTTTGTGAATGGTCGT

P1110

ERCAS12A-
1321
/5Phos/GTTTACACGTACGGGGTACGCATCNNKCGTCGTTTTGTGAATGGTCGTTTT

P1111

ERCAS12A-
1322
/5Phos/TACACGTACGGGGTACGCATCAAANNKCGTTTTGTGAATGGTCGTTTTAGT

P1112

ERCAS12A-
1323
/5Phos/ACGTACGGGGTACGCATCAAACGTNNKTTTGTGAATGGTCGTTTTAGTAAT

P1113

ERCAS12A-
1324
/5Phos/TACGGGGTACGCATCAAACGTCGTNNKGTGAATGGTCGTTTTAGTAATGAG

P1114

ERCAS12A-
1325
/5Phos/GGGGTACGCATCAAACGTCGTTTTNNKAATGGTCGTTTTAGTAATGAGTCG

P1115

ERCAS12A-
1326
/5Phos/GTACGCATCAAACGTCGTTTTGTGNNKGGTCGTTTTAGTAATGAGTCGGAT

P1116

ERCAS12A-
1327
/5Phos/CGCATCAAACGTCGTTTTGTGAATNNKCGTTTTAGTAATGAGTCGGATACA

P1117

ERCAS12A-
1328
/5Phos/ATCAAACGTCGTTTTGTGAATGGTNNKTTTAGTAATGAGTCGGATACAATT

P1118

ERCAS12A-
1329
/5Phos/AAACGTCGTTTTGTGAATGGTCGTNNKAGTAATGAGTCGGATACAATTGAT

P1119

ERCAS12A-
1330
/5Phos/CGTCGTTTTGTGAATGGTCGTTTTNNKAATGAGTCGGATACAATTGATATT

P1120

ERCAS12A-
1331
/5Phos/CGTTTTGTGAATGGTCGTTTTAGTNNKGAGTCGGATACAATTGATATTACC

P1121

ERCAS12A-
1332
/5Phos/TTTGTGAATGGTCGTTTTAGTAATNNKTCGGATACAATTGATATTACCAAG

P1122

ERCAS12A-
1333
/5Phos/GTGAATGGTCGTTTTAGTAATGAGNNKGATACAATTGATATTACCAAGGAT

P1123

ERCAS12A-
1334
/5Phos/AATGGTCGTTTTAGTAATGAGTCGNNKACAATTGATATTACCAAGGATATG

P1124

ERCAS12A-
1335
/5Phos/GGTCGTTTTAGTAATGAGTCGGATNNKATTGATATTACCAAGGATATGGAG

P1125

ERCAS12A-
1336
/5Phos/CGTTTTAGTAATGAGTCGGATACANNKGATATTACCAAGGATATGGAGAAG

P1126

ERCAS12A-
1337
/5Phos/TTTAGTAATGAGTCGGATACAATTNNKATTACCAAGGATATGGAGAAGACT

P1127

ERCAS12A-
1338
/5Phos/AGTAATGAGTCGGATACAATTGATNNKACCAAGGATATGGAGAAGACTCTG

P1128

ERCAS12A-
1339
/5Phos/AATGAGTCGGATACAATTGATATTNNKAAGGATATGGAGAAGACTCTGGAA

P1129

ERCAS12A-
1340
/5Phos/GAGTCGGATACAATTGATATTACCNNKGATATGGAGAAGACTCTGGAAATG

P1130

ERCAS12A-
1341
/5Phos/TCGGATACAATTGATATTACCAAGNNKATGGAGAAGACTCTGGAAATGACA

P1131

ERCAS12A-
1342
/5Phos/GATACAATTGATATTACCAAGGATNNKGAGAAGACTCTGGAAATGACAGAC

P1132

ERCAS12A-
1343
/5Phos/ACAATTGATATTACCAAGGATATGNNKAAGACTCTGGAAATGACAGACATC

P1133

ERCAS12A-
1344
/5Phos/ATTGATATTACCAAGGATATGGAGNNKACTCTGGAAATGACAGACATCAAT

P1134

ERCAS12A-
1345
/5Phos/GATATTACCAAGGATATGGAGAAGNNKCTGGAAATGACAGACATCAATTGG

P1135

ERCAS12A-
1346
/5Phos/ATTACCAAGGATATGGAGAAGACTNNKGAAATGACAGACATCAATTGGCGT

P1136

ERCAS12A-
1347
/5Phos/ACCAAGGATATGGAGAAGACTCTGNNKATGACAGACATCAATTGGCGTGAT

P1137

ERCAS12A-
1348
/5Phos/AAGGATATGGAGAAGACTCTGGAANNKACAGACATCAATTGGCGTGATGGA

P1138

ERCAS12A-
1349
/5Phos/GATATGGAGAAGACTCTGGAAATGNNKGACATCAATTGGCGTGATGGACAT

P1139

ERCAS12A-
1350
/5Phos/ATGGAGAAGACTCTGGAAATGACANNKATCAATTGGCGTGATGGACATGAT

P1140

ERCAS12A-
1351
/5Phos/GAGAAGACTCTGGAAATGACAGACNNKAATTGGCGTGATGGACATGATTTG

P1141

ERCAS12A-
1352
/5Phos/AAGACTCTGGAAATGACAGACATCNNKTGGCGTGATGGACATGATTTGCGT

P1142

ERCAS12A-
1353
/5Phos/ACTCTGGAAATGACAGACATCAATNNKCGTGATGGACATGATTTGCGTCAG

P1143

ERCAS12A-
1354
/5Phos/CTGGAAATGACAGACATCAATTGGNNKGATGGACATGATTTGCGTCAGGAC

P1144

ERCAS12A-
1355
/5Phos/GAAATGACAGACATCAATTGGCGTNNKGGACATGATTTGCGTCAGGACATT

P1145

ERCAS12A-
1356
/5Phos/ATGACAGACATCAATTGGCGTGATNNKCATGATTTGCGTCAGGACATTATC

P1146

ERCAS12A-
1357
/5Phos/ACAGACATCAATTGGCGTGATGGANNKGATTTGCGTCAGGACATTATCGAC

P1147

ERCAS12A-
1358
/5Phos/GACATCAATTGGCGTGATGGACATNNKTTGCGTCAGGACATTATCGACTAT

P1148

ERCAS12A-
1359
/5Phos/ATCAATTGGCGTGATGGACATGATNNKCGTCAGGACATTATCGACTATGAG

P1149

ERCAS12A-
1360
/5Phos/AATTGGCGTGATGGACATGATTTGNNKCAGGACATTATCGACTATGAGATT

P1150

ERCAS12A-
1361
/5Phos/TGGCGTGATGGACATGATTTGCGTNNKGACATTATCGACTATGAGATTGTG

P1151

ERCAS12A-
1362
/5Phos/CGTGATGGACATGATTTGCGTCAGNNKATTATCGACTATGAGATTGTGCAG

P1152

ERCAS12A-
1363
/5Phos/GATGGACATGATTTGCGTCAGGACNNKATCGACTATGAGATTGTGCAGCAT

P1153

ERCAS12A-
1364
/5Phos/GGACATGATTTGCGTCAGGACATTNNKGACTATGAGATTGTGCAGCATATT

P1154

ERCAS12A-
1365
/5Phos/CATGATTTGCGTCAGGACATTATCNNKTATGAGATTGTGCAGCATATTTTC

P1155

ERCAS12A-
1366
/5Phos/GATTTGCGTCAGGACATTATCGACNNKGAGATTGTGCAGCATATTTTCGAA

P1156

ERCAS12A-
1367
/5Phos/TTGCGTCAGGACATTATCGACTATNNKATTGTGCAGCATATTTTCGAAATT

P1157

ERCAS12A-
1368
/5Phos/CGTCAGGACATTATCGACTATGAGNNKGTGCAGCATATTTTCGAAATTTTT

P1158

ERCAS12A-
1369
/5Phos/CAGGACATTATCGACTATGAGATTNNKCAGCATATTTTCGAAATTTTTCGC

P1159

ERCAS12A-
1370
/5Phos/GACATTATCGACTATGAGATTGTGNNKCATATTTTCGAAATTTTTCGCCTG

P1160

ERCAS12A-
1371
/5Phos/ATTATCGACTATGAGATTGTGCAGNNKATTTTCGAAATTTTTCGCCTGACG

P1161

ERCAS12A-
1372
/5Phos/ATCGACTATGAGATTGTGCAGCATNNKTTCGAAATTTTTCGCCTGACGGTG

P1162

ERCAS12A-
1373
/5Phos/GACTATGAGATTGTGCAGCATATTNNKGAAATTTTTCGCCTGACGGTGCAA

P1163

ERCAS12A-
1374
/5Phos/TATGAGATTGTGCAGCATATTTTCNNKATTTTTCGCCTGACGGTGCAAATG

P1164

ERCAS12A-
1375
/5Phos/GAGATTGTGCAGCATATTTTCGAANNKTTTCGCCTGACGGTGCAAATGCGT

P1165

ERCAS12A-
1376
/5Phos/ATTGTGCAGCATATTTTCGAAATTNNKCGCCTGACGGTGCAAATGCGTAAT

P1166

ERCAS12A-
1377
/5Phos/GTGCAGCATATTTTCGAAATTTTTNNKCTGACGGTGCAAATGCGTAATTCC

P1167

ERCAS12A-
1378
/5Phos/CAGCATATTTTCGAAATTTTTCGCNNKACGGTGCAAATGCGTAATTCCTTA

P1168

ERCAS12A-
1379
/5Phos/CATATTTTCGAAATTTTTCGCCTGNNKGTGCAAATGCGTAATTCCTTAAGC

P1169

ERCAS12A-
1380
/5Phos/ATTTTCGAAATTTTTCGCCTGACGNNKCAAATGCGTAATTCCTTAAGCGAA

P1170

ERCAS12A-
1381
/5Phos/TTCGAAATTTTTCGCCTGACGGTGNNKATGCGTAATTCCTTAAGCGAACTG

P1171

ERCAS12A-
1382
/5Phos/GAAATTTTTCGCCTGACGGTGCAANNKCGTAATTCCTTAAGCGAACTGGAA

P1172

ERCAS12A-
1383
/5Phos/ATTTTTCGCCTGACGGTGCAAATGNNKAATTCCTTAAGCGAACTGGAAGAT

P1173

ERCAS12A-
1384
/5Phos/TTTCGCCTGACGGTGCAAATGCGTNNKTCCTTAAGCGAACTGGAAGATCGC

P1174

ERCAS12A-
1385
/5Phos/CGCCTGACGGTGCAAATGCGTAATNNKTTAAGCGAACTGGAAGATCGCGAC

P1175

ERCAS12A-
1386
/5Phos/CTGACGGTGCAAATGCGTAATTCCNNKAGCGAACTGGAAGATCGCGACTAT

P1176

ERCAS12A-
1387
/5Phos/ACGGTGCAAATGCGTAATTCCTTANNKGAACTGGAAGATCGCGACTATGAC

P1177

ERCAS12A-
1388
/5Phos/GTGCAAATGCGTAATTCCTTAAGCNNKCTGGAAGATCGCGACTATGACCGC

P1178

ERCAS12A-
1389
/5Phos/CAAATGCGTAATTCCTTAAGCGAANNKGAAGATCGCGACTATGACCGCTTA

P1179

ERCAS12A-
1390
/5Phos/ATGCGTAATTCCTTAAGCGAACTGNNKGATCGCGACTATGACCGCTTAATT

P1180

ERCAS12A-
1391
/5Phos/CGTAATTCCTTAAGCGAACTGGAANNKCGCGACTATGACCGCTTAATTTCT

P1181

ERCAS12A-
1392
/5Phos/AATTCCTTAAGCGAACTGGAAGATNNKGACTATGACCGCTTAATTTCTCCT

P1182

ERCAS12A-
1393
/5Phos/TCCTTAAGCGAACTGGAAGATCGCNNKTATGACCGCTTAATTTCTCCTGTG

P1183

ERCAS12A-
1394
/5Phos/TTAAGCGAACTGGAAGATCGCGACNNKGACCGCTTAATTTCTCCTGTGCTG

P1184

ERCAS12A-
1395
/5Phos/AGCGAACTGGAAGATCGCGACTATNNKCGCTTAATTTCTCCTGTGCTGAAT

P1185

ERCAS12A-
1396
/5Phos/GAACTGGAAGATCGCGACTATGACNNKTTAATTTCTCCTGTGCTGAATGAG

P1186

ERCAS12A-
1397
/5Phos/CTGGAAGATCGCGACTATGACCGCNNKATTTCTCCTGTGCTGAATGAGAAC

P1187

ERCAS12A-
1398
/5Phos/GAAGATCGCGACTATGACCGCTTANNKTCTCCTGTGCTGAATGAGAACAAC

P1188

ERCAS12A-
1399
/5Phos/GATCGCGACTATGACCGCTTAATTNNKCCTGTGCTGAATGAGAACAACATC

P1189

ERCAS12A-
1400
/5Phos/CGCGACTATGACCGCTTAATTTCTNNKGTGCTGAATGAGAACAACATCTTC

P1190

ERCAS12A-
1401
/5Phos/GACTATGACCGCTTAATTTCTCCTNNKCTGAATGAGAACAACATCTTCTAT

P1191

ERCAS12A-
1402
/5Phos/TATGACCGCTTAATTTCTCCTGTGNNKAATGAGAACAACATCTTCTATGAC

P1192

ERCAS12A-
1403
/5Phos/GACCGCTTAATTTCTCCTGTGCTGNNKGAGAACAACATCTTCTATGACTCA

P1193

ERCAS12A-
1404
/5Phos/CGCTTAATTTCTCCTGTGCTGAATNNKAACAACATCTTCTATGACTCAGCG

P1194

ERCAS12A-
1405
/5Phos/TTAATTTCTCCTGTGCTGAATGAGNNKAACATCTTCTATGACTCAGCGAAA

P1195

ERCAS12A-
1406
/5Phos/ATTTCTCCTGTGCTGAATGAGAACNNKATCTTCTATGACTCAGCGAAAGCC

P1196

ERCAS12A-
1407
/5Phos/TCTCCTGTGCTGAATGAGAACAACNNKTTCTATGACTCAGCGAAAGCCGGG

P1197

ERCAS12A-
1408
/5Phos/CCTGTGCTGAATGAGAACAACATCNNKTATGACTCAGCGAAAGCCGGGGAC

P1198

ERCAS12A-
1409
/5Phos/GTGCTGAATGAGAACAACATCTTCNNKGACTCAGCGAAAGCCGGGGACGCT

P1199

ERCAS12A-
1410
/5Phos/CTGAATGAGAACAACATCTTCTATNNKTCAGCGAAAGCCGGGGACGCTTTG

P1200

ERCAS12A-
1411
/5Phos/AATGAGAACAACATCTTCTATGACNNKGCGAAAGCCGGGGACGCTTTGCCA

P1201

ERCAS12A-
1412
/5Phos/GAGAACAACATCTTCTATGACTCANNKAAAGCCGGGGACGCTTTGCCAAAG

P1202

ERCAS12A-
1413
/5Phos/AACAACATCTTCTATGACTCAGCGNNKGCCGGGGACGCTTTGCCAAAGGAT

P1203

ERCAS12A-
1414
/5Phos/AACATCTTCTATGACTCAGCGAAANNKGGGGACGCTTTGCCAAAGGATGCA

P1204

ERCAS12A-
1415
/5Phos/ATCTTCTATGACTCAGCGAAAGCCNNKGACGCTTTGCCAAAGGATGCAGAT

P1205

ERCAS12A-
1416
/5Phos/TTCTATGACTCAGCGAAAGCCGGGNNKGCTTTGCCAAAGGATGCAGATGCT

P1206

ERCAS12A-
1417
/5Phos/TATGACTCAGCGAAAGCCGGGGACNNKTTGCCAAAGGATGCAGATGCTAAC

P1207

ERCAS12A-
1418
/5Phos/GACTCAGCGAAAGCCGGGGACGCTNNKCCAAAGGATGCAGATGCTAACGGT

P1208

ERCAS12A-
1419
/5Phos/TCAGCGAAAGCCGGGGACGCTTTGNNKAAGGATGCAGATGCTAACGGTGCA

P1209

ERCAS12A-
1420
/5Phos/GCGAAAGCCGGGGACGCTTTGCCANNKGATGCAGATGCTAACGGTGCATAT

P1210

ERCAS12A-
1421
/5Phos/AAAGCCGGGGACGCTTTGCCAAAGNNKGCAGATGCTAACGGTGCATATTGT

P1211

ERCAS12A-
1422
/5Phos/GCCGGGGACGCTTTGCCAAAGGATNNKGATGCTAACGGTGCATATTGTATT

P1212

ERCAS12A-
1423
/5Phos/GGGGACGCTTTGCCAAAGGATGCANNKGCTAACGGTGCATATTGTATTGCT

P1213

ERCAS12A-
1424
/5Phos/GACGCTTTGCCAAAGGATGCAGATNNKAACGGTGCATATTGTATTGCTCTG

P1214

ERCAS12A-
1425
/5Phos/GCTTTGCCAAAGGATGCAGATGCTNNKGGTGCATATTGTATTGCTCTGAAA

P1215

ERCAS12A-
1426
/5Phos/TTGCCAAAGGATGCAGATGCTAACNNKGCATATTGTATTGCTCTGAAAGGC

P1216

ERCAS12A-
1427
/5Phos/CCAAAGGATGCAGATGCTAACGGTNNKTATTGTATTGCTCTGAAAGGCCTT

P1217

ERCAS12A-
1428
/5Phos/AAGGATGCAGATGCTAACGGTGCANNKTGTATTGCTCTGAAAGGCCTTTAC

P1218

ERCAS12A-
1429
/5Phos/GATGCAGATGCTAACGGTGCATATNNKATTGCTCTGAAAGGCCTTTACGAG

P1219

ERCAS12A-
1430
/5Phos/GCAGATGCTAACGGTGCATATTGTNNKGCTCTGAAAGGCCTTTACGAGATC

P1220

ERCAS12A-
1431
/5Phos/GATGCTAACGGTGCATATTGTATTNNKCTGAAAGGCCTTTACGAGATCAAA

P1221

ERCAS12A-
1432
/5Phos/GCTAACGGTGCATATTGTATTGCTNNKAAAGGCCTTTACGAGATCAAACAA

P1222

ERCAS12A-
1433
/5Phos/AACGGTGCATATTGTATTGCTCTGNNKGGCCTTTACGAGATCAAACAAATT

P1223

ERCAS12A-
1434
/5Phos/GGTGCATATTGTATTGCTCTGAAANNKCTTTACGAGATCAAACAAATTACT

P1224

ERCAS12A-
1435
/5Phos/GCATATTGTATTGCTCTGAAAGGCNNKTACGAGATCAAACAAATTACTGAA

P1225

ERCAS12A-
1436
/5Phos/TATTGTATTGCTCTGAAAGGCCTTNNKGAGATCAAACAAATTACTGAAAAC

P1226

ERCAS12A-
1437
/5Phos/TGTATTGCTCTGAAAGGCCTTTACNNKATCAAACAAATTACTGAAAACTGG

P1227

ERCAS12A-
1438
/5Phos/ATTGCTCTGAAAGGCCTTTACGAGNNKAAACAAATTACTGAAAACTGGAAG

P1228

ERCAS12A-
1439
/5Phos/GCTCTGAAAGGCCTTTACGAGATCNNKCAAATTACTGAAAACTGGAAGGAA

P1229

ERCAS12A-
1440
/5Phos/CTGAAAGGCCTTTACGAGATCAAANNKATTACTGAAAACTGGAAGGAAGAT

P1230

ERCAS12A-
1441
/5Phos/AAAGGCCTTTACGAGATCAAACAANNKACTGAAAACTGGAAGGAAGATGGA

P1231

ERCAS12A-
1442
/5Phos/GGCCTTTACGAGATCAAACAAATTNNKGAAAACTGGAAGGAAGATGGAAAG

P1232

ERCAS12A-
1443
/5Phos/CTTTACGAGATCAAACAAATTACTNNKAACTGGAAGGAAGATGGAAAGTTT

P1233

ERCAS12A-
1444
/5Phos/TACGAGATCAAACAAATTACTGAANNKTGGAAGGAAGATGGAAAGTTTTCC

P1234

ERCAS12A-
1445
/5Phos/GAGATCAAACAAATTACTGAAAACNNKAAGGAAGATGGAAAGTTTTCCCGT

P1235

ERCAS12A-
1446
/5Phos/ATCAAACAAATTACTGAAAACTGGNNKGAAGATGGAAAGTTTTCCCGTGAT

P1236

ERCAS12A-
1447
/5Phos/AAACAAATTACTGAAAACTGGAAGNNKGATGGAAAGTTTTCCCGTGATAAA

P1237

ERCAS12A-
1448
/5Phos/CAAATTACTGAAAACTGGAAGGAANNKGGAAAGTTTTCCCGTGATAAATTG

P1238

ERCAS12A-
1449
/5Phos/ATTACTGAAAACTGGAAGGAAGATNNKAAGTTTTCCCGTGATAAATTGAAG

P1239

ERCAS12A-
1450
/5Phos/ACTGAAAACTGGAAGGAAGATGGANNKTTTTCCCGTGATAAATTGAAGATC

P1240

ERCAS12A-
1451
/5Phos/GAAAACTGGAAGGAAGATGGAAAGNNKTCCCGTGATAAATTGAAGATCTCT

P1241

ERCAS12A-
1452
/5Phos/AACTGGAAGGAAGATGGAAAGTTTNNKCGTGATAAATTGAAGATCTCTAAT

P1242

ERCAS12A-
1453
/5Phos/TGGAAGGAAGATGGAAAGTTTTCCNNKGATAAATTGAAGATCTCTAATAAG

P1243

ERCAS12A-
1454
/5Phos/AAGGAAGATGGAAAGTTTTCCCGTNNKAAATTGAAGATCTCTAATAAGGAC

P1244

ERCAS12A-
1455
/5Phos/GAAGATGGAAAGTTTTCCCGTGATNNKTTGAAGATCTCTAATAAGGACTGG

P1245

ERCAS12A-
1456
/5Phos/GATGGAAAGTTTTCCCGTGATAAANNKAAGATCTCTAATAAGGACTGGTTT

P1246

ERCAS12A-
1457
/5Phos/GGAAAGTTTTCCCGTGATAAATTGNNKATCTCTAATAAGGACTGGTTTGAC

P1247

ERCAS12A-
1458
/5Phos/AAGTTTTCCCGTGATAAATTGAAGNNKTCTAATAAGGACTGGTTTGACTTT

P1248

ERCAS12A-
1459
/5Phos/TTTTCCCGTGATAAATTGAAGATCNNKAATAAGGACTGGTTTGACTTTATC

P1249

ERCAS12A-
1460
/5Phos/TCCCGTGATAAATTGAAGATCTCTNNKAAGGACTGGTTTGACTTTATCCAA

P1250

ERCAS12A-
1461
/5Phos/CGTGATAAATTGAAGATCTCTAATNNKGACTGGTTTGACTTTATCCAAAAT

P1251

ERCAS12A-
1462
/5Phos/GATAAATTGAAGATCTCTAATAAGNNKTGGTTTGACTTTATCCAAAATAAA

P1252

ERCAS12A-
1463
/5Phos/AAATTGAAGATCTCTAATAAGGACNNKTTTGACTTTATCCAAAATAAACGT

P1253

ERCAS12A-
1464
/5Phos/TTGAAGATCTCTAATAAGGACTGGNNKGACTTTATCCAAAATAAACGTTAT

P1254

ERCAS12A-
1465
/5Phos/AAGATCTCTAATAAGGACTGGTTTNNKTTTATCCAAAATAAACGTTATTTA

P1255

ERCAS12A-
1466
/5Phos/ATCTCTAATAAGGACTGGTTTGACNNKATCCAAAATAAACGTTATTTAGGT

P1256

ERCAS12A-
1467
/5Phos/TCTAATAAGGACTGGTTTGACTTTNNKCAAAATAAACGTTATTTAGGTCGT

P1257

ERCAS12A-
1468
/5Phos/AATAAGGACTGGTTTGACTTTATCNNKAATAAACGTTATTTAGGTCGTAGC

P1258

ERCAS12A-
1469
/5Phos/AAGGACTGGTTTGACTTTATCCAANNKAAACGTTATTTAGGTCGTAGCAGT

P1259

ERCAS12A-
1470
/5Phos/GACTGGTTTGACTTTATCCAAAATNNKCGTTATTTAGGTCGTAGCAGTGAT

P1260

ERCAS12A-
1471
/5Phos/TGGTTTGACTTTATCCAAAATAAANNKTATTTAGGTCGTAGCAGTGATGAT

P1261

ERCAS12A-
1472
/5Phos/TTTGACTTTATCCAAAATAAACGTNNKTTAGGTCGTAGCAGTGATGATGAA

P1262

ERCAS12A-
1473
/5Phos/GACTTTATCCAAAATAAACGTTATNNKGGTCGTAGCAGTGATGATGAAGCA

P1263

Example 3
Editing Efficiency of ErCas12a Variants in Human Cells

This example evaluates the DNA cleavage activity of 20 ErCas12a point mutations in the context of E. coli (FIG. 7). The cleavage activity of each mutant was quantified by calculating the survival rate of E. coli with or without the induction of toxic reporter gene. Compared to WT-ERCAS12A, 31 out of 40 point mutations showed improved DNA cleavage, thus further validating the result of sequencing based, high-throughput phenotype measurement in Example 1.

Eight representative ERCAS12A point mutations were selected for further characterization in human cells. These point mutations included 135L, G39Q, 140Q, S469K, Y507S, K516P, T826F and Y907V. Site directed mutagenesis was performed to introduce specific point mutations on the expression vector using primers listed in Table 4. Each protein was expressed in E. coli BL21DE3 cells, sequentially purified using nickel-affinity and heparin chromatography. To evaluate the genome editing efficiency of each variant, ERCAS12A-RNPs were assembled using 6 crRNAs targeting human HPRT gene (Table 5). The assembled RNPs (50 nM or 200 nM) were delivered into HEK293 cells by Lonza nucleofection (SF buffer, DS-150 program), and the editing efficiency was measured by T7 endonuclease I assay (T7EI) 48-hours post delivery (FIG. 8A-B). To facilitate the comparison, the efficiency of variant at each target was normalized to WT-ERCAS12A and presented as fold-improvement over WT (FIG. 8C-D). The single mutant variants showed improved activity over WT at one or multiple sites, especially at low RNP concentration (50 nM, FIG. 8B, D).

TABLE 4

Primers for Site Directed Mutagenesis of ErCas12a

SEQ ID

Mutation
NO
DNA Primers (5′→3′)

I35L Fwd
1474
ACTGAGACAACACAGCAGTTCCTGGTTAAAAACGGCATCATTAAAGAAGATG

I35L Rev
1475
CATCTTCTTTAATGATGCCGTTTTTAACCAGGAACTGCTGTGTTGTCTCAGT

G39Q Fwd
1476
CACAGCAGTTCATCGTTAAAAACCAGATCATTAAAGAAGATGAGCTGCG

G39Q Rev
1477
CGCAGCTCATCTTCTTTAATGATCTGGTTTTTAACGATGAACTGCTGTG

I40Q Fwd
1478
CAGTTCATCGTTAAAAACGGCCAGATTAAAGAAGATGAGCTGCGC

I40Q Rev
1479
GCGCAGCTCATCTTCTTTAATCTGGCCGTTTTTAACGATGAACTG

S469K Fwd
1480
ACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTAGTAGA

S469K Rev
1481
TCTACTAACTCCTCTGTCATAAACACTTTGCACCAATGGAAGGCGT

Y507S Fwd
1482
GATTTCACTGTATAACTTAGTCCGTAATAGCGTCACTCAGAAGCCGTACTC

Y507S Rev
1483
GAGTACGGCTTCTGAGTGACGCTATTACGGACTAAGTTATACAGTGAAATC

K516P Fwd
1484
TCAGAAGCCGTACTCAACTCCGAAGATCAAGTTAAATTTTGGAATTCCGA

K516P Rev
1485
TCGGAATTCCAAAATTTAACTTGATCTTCGGAGTTGAGTACGGCTTCTGA

T826F Fwd
1486
GTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTAC

T826F Rev
1487
GTAGCGATAGTCTTTCACAATGTTAAATGCTGCTTCGTGGTGAC

Y907V Fwd
1488
AGCAAAAGTCTTTTAATATCGTTAATGGTGTGGACTACCAAATTAAGCTTAAACAA

CAGG

Y907V Rev
1489
CCTGTTGTTTAAGCTTAATTTGGTAGTCCACACCATTAACGATATTAAAAGACTTT

TGCT

TABLE 5

crRNA and Primer Sequences for T7E1 Assay

SEQ ID

crRNA
NO
crRNA RNA Sequences (5′→3′)

crRNA-1
1490
UUAAUUUCUACUCUUGUAGAUCUUGGGUGUGUUAAAAGUGAC

crRNA-2
1491
UUAAUUUCUACUCUUGUAGAUAUCCGUGCUGAGUGUACCAUG

crRNA-3
1492
UUAAUUUCUACUCUUGUAGAUUAAUGCCCUGUAGUCUCUCUG

crRNA-4
1493
UUAAUUUCUACUCUUGUAGAUUAAUUAACAGCUUGCUGGUGA

crRNA-5
1494
UUAAUUUCUACUCUUGUAGAUUGUGAAAUGGCUUAUAAUUGC

crRNA-6
1495
UUAAUUUCUACUCUUGUAGAUUUGUAGGAUAUGCCCUUGACU

DNA Primer Sequences (5′→3′)

HPRT7 Fwd
1496
AAGAATGTTGTGATAAAAGGTGATGCT

HPRT7 Rev
1497
ACACATCCATGGGACTTCTGCCTC

Example 4
Editing Efficiency of ErCas12 Variants in Human Cells

This example evaluates the editing efficiency of 42 additional ErCas12a point mutations selected based on previous E. coli screen data. Site directed mutagenesis was performed to introduce specific point mutations on the expression vector using primers listed in Table 6. Purified plasmids were lipofected into human HEK293 cells with 2 crRNAs targeting human HPRT gene (Table 7). The editing efficiency of HPRT was measured by T7EI assay 2 days post-delivery. To facilitate the comparison, the efficiency of variant at each target was normalized to WT ERCAS12A and presented as fold-improvement over WT (FIG. 9). 21 of 42 mutants improved the on-targeting efficiency of ERCAS12A when delivered as plasmid (FIG. 8, Table 7). Four point mutations, including S158G, D303R, F465R, and F840L, were selected for further characterization as RNP. 3 out of 4 mutations (D303R, F465R, F840L) improved the on-target editing efficiency in nearly all target sites (FIG. 8B-C).

TABLE 6

Primers for Site Directed Mutagenesis of ERCAS12A

SEQ ID

Mutation
NO
Fwd Primers (5′→3′)

Q18V Fwd
1498
ATCGGCATCAGCAGCCTGGTGAAAACCCTGCGGAACGCT

Q18V Rev
1499
AGCGTTCCGCAGGGTTTTCACCAGGCTGCTGATGCCGAT

D56R Fwd
1500
CGAGAACCGGCAGATCCTGAAGCGGATCATGGACGACTACTACCGGG

D56R Rev
1501
CCCGGTAGTAGTCGTCCATGATCCGCTTCAGGATCTGCCGGTTCTCG

D59A Fwd
1502
GCAGATCCTGAAGGACATCATGGCCGACTACTACCGGGGCTTCATCA

D59A Rev
1503
TGATGAAGCCCCGGTAGTAGTCGGCCATGATGTCCTTCAGGATCTGC

S158G Fwd
1504
AGAAAACCCAAGTCATCAAGCTGTTCGGCAGATTCGCCACCAGCTTCAA

S158G Rev
1505
TTGAAGCTGGTGGCGAATCTGCCGAACAGCTTGATGACTTGGGTTTTCT

K165Y Fwd
1506
GCAGATTCGCCACCAGCTTCTACGACTACTTCAAGAACCGGGCCAAC

K165Y Rev
1507
GTTGGCCCGGTTCTTGAAGTAGTCGTAGAAGCTGGTGGCGAATCTGC

D178R Fwd
1508
GCCAACTGCTTCAGCGCCCGGGATATCAGCAGCAGCTCCTGC

D178R Rev
1509
GCAGGAGCTGCTGCTGATATCCCGGGCGCTGAAGCAGTTGGC

S181V Fwd
1510
GCTTCAGCGCCGACGATATCGTGAGCAGCTCCTGCCACAGA

S181V Rev
1511
TCTGTGGCAGGAGCTGCTCACGATATCGTCGGCGCTGAAGC

A290K Fwd
1512
GCTGCACAAGCAGATTCTGTGTATCAAGGACACCTCCTACGAGGTGCC

A290K Rev
1513
GGCACCTCGTAGGAGGTGTCCTTGATACACAGAATCTGCTTGTGCAGC

V296R Fwd
1514
TCGCCGACACCTCCTACGAGCGGCCCTACAAGTTCGAGAGCGAC

V296R Rev
1515
GTCGCTCTCGAACTTGTAGGGCCGCTCGTAGGAGGTGTCGGCGA

P297V Fwd
1516
CCGACACCTCCTACGAGGTGGTGTACAAGTTCGAGAGCGACGAAGAG

P297V Rev
1517
CTCTTCGTCGCTCTCGAACTTGTACACCACCTCGTAGGAGGTGTCGG

P297R Fwd
1518
CCGACACCTCCTACGAGGTGCGGTACAAGTTCGAGAGCGACGAAGAG

P297R Rev
1519
CTCTTCGTCGCTCTCGAACTTGTACCGCACCTCGTAGGAGGTGTCGG

Y298R Fwd
1520
CACCTCCTACGAGGTGCCCCGGAAGTTCGAGAGCGACGAAGAGG

Y298R Rev
1521
CCTCTTCGTCGCTCTCGAACTTCCGGGGCACCTCGTAGGAGGTG

D303R Fwd
1522
GTGCCCTACAAGTTCGAGAGCCGGGAAGAGGTGTACCAGAGCGT

D303R Rev
1523
ACGCTCTGGTACACCTCTTCCCGGCTCTCGAACTTGTAGGGCAC

E305K Fwd
1524
CCTACAAGTTCGAGAGCGACGAAAAGGTGTACCAGAGCGTGAAC

E305K Rev
1525
GTTCACGCTCTGGTACACCTTTTCGTCGCTCTCGAACTTGTAGG

Y307K Fwd
1526
TTCGAGAGCGACGAAGAGGTGAAGCAGAGCGTGAACGGCTTC

Y307K Rev
1527
GAAGCCGTTCACGCTCTGCTTCACCTCTTCGTCGCTCTCGAA

Q353R Fwd
1528
AAGTTCTACGAGAGCGTGTCCCGGAAAACCTACAGAGACTGGGAGAC

Q353R Rev
1529
GTCTCCCAGTCTCTGTAGGTTTTCCGGGACACGCTCTCGTAGAACTT

F465R Fwd
1530
TGGACGTGATCATGAACGCCCGGCACTGGTGCAGCGTGTTC

F465R Rev
1531
GAACACGCTGCACCAGTGCCGGGCGTTCATGATCACGTCCA

V470K Fwd
1532
GCCTTCCACTGGTGCAGCAAGTTCATGACCGAGGAACTGGTG

V470K Rev
1533
CACCAGTTCCTCGGTCATGAACTTGCTGCACCAGTGGAAGGC

Y507R Fwd
1534
GTACAACCTCGTGCGGAACCGGGTGACCCAGAAGCCTTACAG

Y507R Rev
1535
CTGTAAGGCTTCTGGGTCACCCGGTTCCGCACGAGGTTGTAC

P512V Fwd
1536
CGGAACTACGTGACCCAGAAGGTGTACAGCACCAAGAAGATCAAGCT

P512V Rev
1537
AGCTTGATCTTCTTGGTGCTGTACACCTTCTGGGTCACGTAGTTCCG

Y513R Fwd
1538
CTACGTGACCCAGAAGCCTCGGAGCACCAAGAAGATCAAGCTG

Y513R Rev
1539
CAGCTTGATCTTCTTGGTGCTCCGAGGCTTCTGGGTCACGTAG

S514R Fwd
1540
CTACGTGACCCAGAAGCCTTACCGGACCAAGAAGATCAAGCTGAACTTC

S514R Rev
1541
GAAGTTCAGCTTGATCTTCTTGGTCCGGTAAGGCTTCTGGGTCACGTAG

K516R Fwd
1542
CCCAGAAGCCTTACAGCACCCGGAAGATCAAGCTGAACTTCGGC

K516R Rev
1543
GCCGAAGTTCAGCTTGATCTTCCGGGTGCTGTAAGGCTTCTGGG

G553M Fwd
1544
GCGGGACAATCTGTACTACCTGATGATCTTCAACGCCAAGAACAAGC

G553M Rev
1545
GCTTGTTCTTGGCGTTGAAGATCATCAGGTAGTACAGATTGTCCCGC

A541K Fwd
1546
AAGAGCAAAGAGTACAGCAACAACGTGATCATCCTGATGCGGGACAA

A541K Rev
1547
TTGTCCCGCATCAGGATGATCACGTTGTTGCTGTACTCTTTGCTCTT

D562K Fwd
1548
TCAACGCCAAGAACAAGCCCAAGAAGAAGATTATCGAGGGCAACAC

D562K Rev
1549
GTGTTGCCCTCGATAATCTTCTTCTTGGGCTTGTTCTTGGCGTTGA

N583V Fwd
1550
GGCGACTACAAGAAGATGATCTATGTGCTGCTGCCTGGGCCG

N583V Rev
1551
CGGCCCAGGCAGCAGCACATAGATCATCTTCTTGTAGTCGCC

L585K Fwd
1552
GCGACTACAAGAAGATGATCTATAACCTGAAGCCTGGGCCGAACAAGATG

L585K Rev
1553
CATCTTGTTCGGCCCAGGCTTCAGGTTATAGATCATCTTCTTGTAGTCGC

F653R Fwd
1554
CCATCCATCCAGAGTGGAAGAATCGGGGCTTCGACTTCAGCGAC

F653R Rev
1555
GTCGCTGAAGTCGAAGCCCCGATTCTTCCACTCTGGATGGATGG

K712R Fwd
1556
CAGATCTACAACAAAGACTTCAGCAAGCGGTCCACCGGCAACGACAA

K712R Rev
1557
TTGTCGTTGCCGGTGGACCGCTTGCTGAAGTCTTTGTTGTAGATCTG

N716R Fwd
1558
AGCAAGAAGTCCACCGGCCGGGACAACCTGCACACCATGTAC

N716R Rev
1559
GTACATGGTGTGCAGGTTGTCCCGGCCGGTGGACTTCTTGCT

A744N Fwd
1560
CTGAAGCTGAACGGCGAGAACGAGATTTTTTTCAGAAAGTCCAGCATC

A744N Rev
1561
GATGCTGGACTTTCTGAAAAAAATCTCGTTCTCGCCGTTCAGCTTCAG

A744T Fwd
1562
CTGAAGCTGAACGGCGAGACCGAGATTTTTTTCAGAAAGTCCAGCATC

A744T Rev
1563
GATGCTGGACTTTCTGAAAAAAATCTCGGTCTCGCCGTTCAGCTTCAG

F748A Fwd
1564
CGGCGAGGCCGAGATTTTTGCCAGAAAGTCCAGCATCAAGAACC

F748A Rev
1565
GGTTCTTGATGCTGGACTTTCTGGCAAAAATCTCGGCCTCGCCG

S751A Fwd
1566
CGAGGCCGAGATTTTTTTCAGAAAGGCCAGCATCAAGAACCCCATCAT

S751A Rev
1567
ATGATGGGGTTCTTGATGCTGGCCTTTCTGAAAAAAATCTCGGCCTCG

I757K Fwd
1568
AGTCCAGCATCAAGAACCCCAAGATCCACAAAAAGGGCAGCAT

I757K Rev
1569
ATGCTGCCCTTTTTGTGGATCTTGGGGTTCTTGATGCTGGACT

E771R Fwd
1570
CATCCTGGTCAACCGGACATACCGGGCCGAAGAGAAGGATCAGTTCG

E771R Rev
1571
CGAACTGATCCTTCTCTTCGGCCCGGTATGTCCGGTTGACCAGGATG

T826K Fwd
1572
CACCACGAAGCCGCCAAGAATATCGTGAAGGACTACCGGT

T826K Rev
1573
ACCGGTAGTCCTTCACGATATTCTTGGCGGCTTCGTGGTG

F840L Fwd
1574
ACTATCGCTACACTTACGATAAGTACCTGTTACATATGCCTATTACGATTAACTT

TAAGG

F840L Rev
1575
CCTTAAAGTTAATCGTAATAGGCATATGTAACAGGTACTTATCGTAAGTGTAGCG

ATAGT

E935R Fwd
1576
GGAAAGAGATCGGCAAGATCAAACGGATCAAAGAGGGCTACCTGAGC

E935R Rev
1577
GCTCAGGTAGCCCTCTTTGATCCGTTTGATCTTGCCGATCTCTTTCC

I936R Fwd
1578
GAAAGAGATCGGCAAGATCAAAGAACGGAAAGAGGGCTACCTGAGCC

I936R Rev
1579
GGCTCAGGTAGCCCTCTTTCCGTTCTTTGATCTTGCCGATCTCTTTC

K1019V Fwd
1580
CTGACCTACATTCCCGACGTGCTGAAAAACGTGGGCCACCA

K1019V Rev
1581
TGGTGGCCCACGTTTTTCAGCACGTCGGGAATGTAGGTCAG

TABLE 7

crRNA and Primer Sequences for T7E1 Assay

SEQ ID

crRNA
NO
crRNA RNA Sequences (5′→3′)

crRNA-1
1490
UUAAUUUCUACUCUUGUAGAUCUUGGGUGUGUUAAAAGUGAC

crRNA-2
1491
UUAAUUUCUACUCUUGUAGAUAUCCGUGCUGAGUGUACCAUG

crRNA-3
1492
UUAAUUUCUACUCUUGUAGAUUAAUGCCCUGUAGUCUCUCUG

crRNA-4
1493
UUAAUUUCUACUCUUGUAGAUUAAUUAACAGCUUGCUGGUGA

crRNA-5
1494
UUAAUUUCUACUCUUGUAGAUUGUGAAAUGGCUUAUAAUUGC

crRNA-6
1495
UUAAUUUCUACUCUUGUAGAUUUGUAGGAUAUGCCCUUGACU

DNA Primer Sequences (5'->3')

HPRT7 Fwd
1496
AAGAATGTTGTGATAAAAGGTGATGCT

HPRT7 Rev
1497
ACACATCCATGGGACTTCTGCCTC

With a large collection of single point mutations with enhanced activity available, mutations were combined or “stacked” to further improve the activity of ERCAS12A. The point mutations described in previous examples were stacked in 9 combinations and the editing efficiency of these ERCAS12A variants were evaluated in human cells as RNPs (FIG. 9A-B). Notably, all stacked variants enhanced the on-target editing efficiency of ERCAS12A over one or multiple target sites. Among these, a variant with double mutations (S469K/T826F) had superior performance.

Example 5
Editing Efficiency of ErCas12 Variants in Human Cell Lines

The editing efficiency of another 20 ERCAS12A point mutations was evaluated. The additional 20 ERCAS12A point mutations were selected based on the E. coli screening data. Site directed mutagenesis was performed to introduce specific point mutations on the expression vector using primers listed in Table 8. Purified plasmids were lipofected into human HEK293 cells with 8 crRNAs targeting human therapeutically relevant genes (Table 9). The editing efficiency of each gene was measured by NGS 3 days post-delivery (FIG. 10A-H).

TABLE 8

Primers for Site Directed Mutagenesis of ERCAS12A

SEQ ID

Mutation
NO
Fwd Primers (5′→3′)

D303C Fwd
1582
GCCCTACAAGTTCGAGAGCTGCGAAGAGGTGTACCAGAGCG

D303C Rev
1583
CGCTCTGGTACACCTCTTCGCAGCTCTCGAACTTGTAGGGC

L444W Fwd
1584
AAGTACAACCCAGAGATCCACTGGGTGGAAAGCGAGCTGAAG

L444W Rev
1585
CTTCAGCTCGCTTTCCACCCAGTGGATCTCTGGGTTGTACTT

Y513G Fwd
1586
CGTGACCCAGAAGCCTGGCAGCACCAAGAAGATCAAGCT

Y513G Rev
1587
AGCTTGATCTTCTTGGTGCTGCCAGGCTTCTGGGTCACG

S514P Fwd
1588
CGTGACCCAGAAGCCTTACCCCACCAAGAAGATCAAGCTGAACT

S514P Rev
1589
AGTTCAGCTTGATCTTCTTGGTGGGGTAAGGCTTCTGGGTCACG

K517P Fwd
1590
GAAGCCTTACAGCACCAAGCCCATCAAGCTGAACTTCGGC

K517P Rev
1591
GCCGAAGTTCAGCTTGATGGGCTTGGTGCTGTAAGGCTTC

I518P Fwd
1592
GCCTTACAGCACCAAGAAGCCCAAGCTGAACTTCGGCATCC

I518P Rev
1593
GGATGCCGAAGTTCAGCTTGGGCTTCTTGGTGCTGTAAGGC

L520M Fwd
1594
CTTACAGCACCAAGAAGATCAAGATGAACTTCGGCATCCCCAC

L520M Rev
1595
GTGGGGATGCCGAAGTTCATCTTGATCTTCTTGGTGCTGTAAG

F522A Fwd
1596
CACCAAGAAGATCAAGCTGAACGCCGGCATCCCCACACTGG

F522A Rev
1597
CCAGTGTGGGGATGCCGGCGTTCAGCTTGATCTTCTTGGTG

N556E Fwd
1598
CTGTACTACCTGGGCATCTTCGAGGCCAAGAACAAGCCCG

N556E Rev
1599
CGGGCTTGTTCTTGGCCTCGAAGATGCCCAGGTAGTACAG

E731P Fwd
1600
GAAGAACCTGTTCAGCGAGCCCAACCTCAAGGACATCGTGC

E731P Rev
1601
GCACGATGTCCTTGAGGTTGGGCTCGCTGAACAGGTTCTTC

N732T Fwd
1602
AACCTGTTCAGCGAGGAAACCCTCAAGGACATCGTGCTGA

N732T Rev
1603
TCAGCACGATGTCCTTGAGGGTTTCCTCGCTGAACAGGTT

A744N Fwd
1604
GAAGCTGAACGGCGAGAACGAGATTTTTTTCAGAAAGTCCAGCA

A744N Rev
1605
TGCTGGACTTTCTGAAAAAAATCTCGTTCTCGCCGTTCAGCTTC

K754G Fwd
1606
GATTTTTTTCAGAAAGTCCAGCATCGGCAACCCCATCATCCACAAAAAG

K754G Rev
1607
CTTTTTGTGGATGATGGGGTTGCCGATGCTGGACTTTCTGAAAAAAATC

I757W Fwd
1608
TCCAGCATCAAGAACCCCTGGATCCACAAAAAGGGCAGCA

I757W Rev
1609
TGCTGCCCTTTTTGTGGATCCAGGGGTTCTTGATGCTGGA

N787A Fwd
1610
CATCCAGATTGTGCGGAAGGCCATCCCCGAGAACATCTATCAAG

N787A Rev
1611
CTTGATAGATGTTCTCGGGGATGGCCTTCCGCACAATCTGGATG

Y832R Fwd
1612
CCACCAATATCGTGAAGGACCGGCGGTATACCTACGATAAGTACTTCC

Y832R Rev
1613
GGAAGTACTTATCGTAGGTATACCGCCGGTCCTTCACGATATTGGTGG

Y836K Fwd
1614
GTGAAGGACTACCGGTATACCAAGGATAAGTACTTCCTGCACATGCC

Y836K Rev
1615
GGCATGTGCAGGAAGTACTTATCCTTGGTATACCGGTAGTCCTTCAC

Y839L Fwd
1616
GACTACCGGTATACCTACGATAAGCTGTTCCTGCACATGCCCA

Y839L Rev
1617
TGGGCATGTGCAGGAACAGCTTATCGTAGGTATACCGGTAGTC

Y907I Fwd
1618
GCTTCAACATCGTGAATGGCATCGACTACCAGATCAAACTGAAGCA

Y907I Rev
1619
TGCTTCAGTTTGATCTGGTAGTCGATGCCATTCACGATGTTGAAGC

K974G Fwd
1620
CTTCAAGAAAGGCCGCTTCGGCGTGGAAAGACAGGTCTACCAG

K974G Rev
1621
CTGGTAGACCTGTCTTTCCACGCCGAAGCGGCCTTTCTTGAAG

D303C Fwd
1582
GCCCTACAAGTTCGAGAGCTGCGAAGAGGTGTACCAGAGCG

D303C Rev
1583
CGCTCTGGTACACCTCTTCGCAGCTCTCGAACTTGTAGGGC

L444W Fwd
1584
AAGTACAACCCAGAGATCCACTGGGTGGAAAGCGAGCTGAAG

L444W Rev
1585
CTTCAGCTCGCTTTCCACCCAGTGGATCTCTGGGTTGTACTT

Y513G Fwd
1586
CGTGACCCAGAAGCCTGGCAGCACCAAGAAGATCAAGCT

Y513G Rev
1587
AGCTTGATCTTCTTGGTGCTGCCAGGCTTCTGGGTCACG

S514P Fwd
1588
CGTGACCCAGAAGCCTTACCCCACCAAGAAGATCAAGCTGAACT

S514P Rev
1589
AGTTCAGCTTGATCTTCTTGGTGGGGTAAGGCTTCTGGGTCACG

K517P Fwd
1590
GAAGCCTTACAGCACCAAGCCCATCAAGCTGAACTTCGGC

K517P Rev
1591
GCCGAAGTTCAGCTTGATGGGCTTGGTGCTGTAAGGCTTC

I518P Fwd
1592
GCCTTACAGCACCAAGAAGCCCAAGCTGAACTTCGGCATCC

I518P Rev
1593
GGATGCCGAAGTTCAGCTTGGGCTTCTTGGTGCTGTAAGGC

L520M Fwd
1594
CTTACAGCACCAAGAAGATCAAGATGAACTTCGGCATCCCCAC

L520M Rev
1595
GTGGGGATGCCGAAGTTCATCTTGATCTTCTTGGTGCTGTAAG

F522A Fwd
1596
CACCAAGAAGATCAAGCTGAACGCCGGCATCCCCACACTGG

F522A Rev
1597
CCAGTGTGGGGATGCCGGCGTTCAGCTTGATCTTCTTGGTG

N556E Fwd
1598
CTGTACTACCTGGGCATCTTCGAGGCCAAGAACAAGCCCG

N556E Rev
1599
CGGGCTTGTTCTTGGCCTCGAAGATGCCCAGGTAGTACAG

E731P Fwd
1600
GAAGAACCTGTTCAGCGAGCCCAACCTCAAGGACATCGTGC

E731P Rev
1601
GCACGATGTCCTTGAGGTTGGGCTCGCTGAACAGGTTCTTC

N732T Fwd
1602
AACCTGTTCAGCGAGGAAACCCTCAAGGACATCGTGCTGA

N732T Rev
1603
TCAGCACGATGTCCTTGAGGGTTTCCTCGCTGAACAGGTT

A744N Fwd
1604
GAAGCTGAACGGCGAGAACGAGATTTTTTTCAGAAAGTCCAGCA

A744N Rev
1605
TGCTGGACTTTCTGAAAAAAATCTCGTTCTCGCCGTTCAGCTTC

K754G Fwd
1606
GATTTTTTTCAGAAAGTCCAGCATCGGCAACCCCATCATCCACAAAAAG

K754G Rev
1607
CTTTTTGTGGATGATGGGGTTGCCGATGCTGGACTTTCTGAAAAAAATC

I757W Fwd
1608
TCCAGCATCAAGAACCCCTGGATCCACAAAAAGGGCAGCA

I757W Rev
1609
TGCTGCCCTTTTTGTGGATCCAGGGGTTCTTGATGCTGGA

N787A Fwd
1610
CATCCAGATTGTGCGGAAGGCCATCCCCGAGAACATCTATCAAG

N787A Rev
1611
CTTGATAGATGTTCTCGGGGATGGCCTTCCGCACAATCTGGATG

Y832R Fwd
1612
CCACCAATATCGTGAAGGACCGGCGGTATACCTACGATAAGTACTTCC

Y832R Rev
1613
GGAAGTACTTATCGTAGGTATACCGCCGGTCCTTCACGATATTGGTGG

Y836K Fwd
1614
GTGAAGGACTACCGGTATACCAAGGATAAGTACTTCCTGCACATGCC

Y836K Rev
1615
GGCATGTGCAGGAAGTACTTATCCTTGGTATACCGGTAGTCCTTCAC

Y839L Fwd
1616
GACTACCGGTATACCTACGATAAGCTGTTCCTGCACATGCCCA

Y839L Rev
1617
TGGGCATGTGCAGGAACAGCTTATCGTAGGTATACCGGTAGTC

Y907I Fwd
1618
GCTTCAACATCGTGAATGGCATCGACTACCAGATCAAACTGAAGCA

Y907I Rev
1619
TGCTTCAGTTTGATCTGGTAGTCGATGCCATTCACGATGTTGAAGC

K974G Fwd
1620
CTTCAAGAAAGGCCGCTTCGGCGTGGAAAGACAGGTCTACCAG

K974G Rev
1621
CTGGTAGACCTGTCTTTCCACGCCGAAGCGGCCTTTCTTGAAG

TABLE 9

crRNA for Editing Experiments and NGS Primer Sequences

SEQ ID

Target
NO
crRNA RNA Sequences (5′→3′)

AAVS1
1622
UUUUUAAUUUCUACUCUUGUAGAUGACCCCUAAGGGAAGAAUGAG

PD1
1623
UUUUUAAUUUCUACUCUUGUAGAUGCACGAAGCUCUCCGAUGUGU

TGFBR2
1624
UUUUUAAUUUCUACUCUUGUAGAUUGAUGUGAGAUUUUCCACCUG

TRAC1
1625
UUUUUAAUUUCUACUCUUGUAGAUCAGAUACGAACCUAAACUUUC

B2M-12
1626
UUUUUAAUUUCUACUCUUGUAGAUUCACAGCCCAAGAUAGUUAAG

CIITA
1627
UUUUUAAUUUCUACUCUUGUAGAUCCGGCCUUUUUACCUUGGGGC

CISH
1628
UUUUUAAUUUCUACUCUUGUAGAUACUGACAGCGUGAACAGGUAG

TRAC2
1629
UUUUUAAUUUCUACUCUUGUAGAUGAGUCUCUCAGCUGGUACACG

SEQ ID

Primer
NO
NGS DNA Primers (5′→3′)

AAVS1 Fwd
1630
acactctttccctacacgacgctcttccgatctGCTGTCACACTCCAGTT

AAVS1 Rev
1631
gtgactggagttcagacgtgtgctcttccgatctACCCAGTGTCCACAAG

PD1 Fwd
1632
acactctttccctacacgacgctcttccgatctAGCTTGTCCGTCTGGTT

PD1 Rev
1633
gtgactggagttcagacgtgtgctcttccgatctAGACTCCCCAGACAGG

TGFBR2 Fwd
1634
acactctttccctacacgacgctcttccgatctCATGATAGTCACTGACAACAAC

G

TGFBR2 Rev
1635
gtgactggagttcagacgtgtgctcttccgatctCTTACCATACAGCCACACAGA

TRAC1 Fwd
1636
acactctttccctacacgacgctcttccgatctACATGCAAGCCCATAACC

TRAC1 Rev
1637
gtgactggagttcagacgtgtgctcttccgatctGGTTTGCTCTCAGAGCTTA

B2M-12 Fwd
1638
acactctttccctacacgacgctcttccgatctCATCCGACATTGAAGTTGACTT

A

B2M-12 Rev
1639
gtgactggagttcagacgtgtgctcttccgatctCTACTCATACACAACTTTCAG

CA

CIITA Fwd
1640
acactctttccctacacgacgctcttccgatctTGGGATTCCTACACAATGC

CIITA Rev
1641
gtgactggagttcagacgtgtgctcttccgatctCCCATCCCCAATTCAGAATG

CISH Fwd
1642
acactctttccctacacgacgctcttccgatctCACAAGGCTGACCACATC

CISH Rev
1643
gtgactggagttcagacgtgtgctcttccgatctCAGAAGGCACGTTCTTAGT

TRAC2 Fwd
1644
acactctttccctacacgacgctcttccgatctATCCTCTTGTCCCACAGATATC

TRAC2 Rev
1645
gtgactggagttcagacgtgtgctcttccgatctCCATAGACCTCATGTCTAGCA

Example 6
Editing Efficiency of ERCAS12A Variants in Human Cells

Evaluation of the editing efficiency of the ERCAS12A Y513G point mutation that was selected from plasmid screening was performed. Site directed mutagenesis was performed to introduce the point mutation on the protein expression vector using primers listed in Table 10. Purified protein was tested as RNP in human HEK293 cells with 7 crRNAs targeting human therapeutically relevant genes (Table 11). The editing efficiency of each gene was measured by NGS 2 days post-delivery (FIG. 11).

Site directed mutagenesis was then used to add additional mutations to ERCAS12A Y513G, Y832R, and F840L, and to make S514P, on the protein expression vector using primers listed in Table 10. Purified protein was tested as RNP in human HEK293 cells with 4 crRNAs targeting human therapeutically relevant genes (Table 11). The editing efficiency of each gene was measured by NGS 2 days post-delivery (FIG. 12A-D). This experiment also included Acidaminococcus sp. Cas12a M537R/F870L (AsCas12a-Ultra) as a positive control, and with and without electroporation enhancer.

TABLE 10

Primers for Site Directed Mutagenesis of ERCAS12A

SEQ ID

Primer
NO
DNA Primers (5′→3′)

Y513G Fwd
1646
ACGTCACTCAGAAGCCGGGCTCAACTAAAAAGATCAAGTTAAATTTTGGAATT

Y513G Rev
1647
AATTCCAAAATTTAACTTGATCTTTTTAGTTGAGCCCGGCTTCTGAGTGACGT

S514P Fwd
1648
CGTCACTCAGAAGCCGTACCCGACTAAAAAGATCAAGTTAAATTTTGGAATTCC

S514P Rev
1649
GGAATTCCAAAATTTAACTTGATCTTTTTAGTCGGGTACGGCTTCTGAGTGACG

Y832R Fwd
1650
CAGCAACAAACATTGTGAAAGACCGTCGCTACACTTACGATAAGTACTTTTTAC

Y832R Rev
1651
GTAAAAAGTACTTATCGTAAGTGTAGCGACGGTCTTTCACAATGTTTGTTGCTG

F840L Fwd
1652
ACTATCGCTACACTTACGATAAGTACCTGTTACATATGCCTATTACGATTAACT

TTAAGG

F840L Rev
1653
CCTTAAAGTTAATCGTAATAGGCATATGTAACAGGTACTTATCGTAAGTGTAGC

GATAGT

TABLE 11

crRNA for Editing Experiments and NGS Primer Sequences

SEQ ID

Target
NO
crRNA RNA Sequences (5′→3′)

AAVS1
1622
UUUUUAAUUUCUACUCUUGUAGAUGACCCCUAAGGGAAGAAUGAG

PD1
1623
UUUUUAAUUUCUACUCUUGUAGAUGCACGAAGCUCUCCGAUGUGU

TGFBR2
1624
UUUUUAAUUUCUACUCUUGUAGAUUGAUGUGAGAUUUUCCACCUG

TRAC1
1625
UUUUUAAUUUCUACUCUUGUAGAUCAGAUACGAACCUAAACUUUC

B2M-12
1626
UUUUUAAUUUCUACUCUUGUAGAUUCACAGCCCAAGAUAGUUAAG

CIITA
1627
UUUUUAAUUUCUACUCUUGUAGAUCCGGCCUUUUUACCUUGGGGC

CISH
1628
UUUUUAAUUUCUACUCUUGUAGAUACUGACAGCGUGAACAGGUAG

TRAC2
1629
UUUUUAAUUUCUACUCUUGUAGAUGAGUCUCUCAGCUGGUACACG

SEQ ID

Primer
NO
NGS DNA Primers (5′→3′)

AAVS1 Fwd
1630
acactctttccctacacgacgctcttccgatctGCTGTCACACTCCAGTT

AAVS1 Rev
1631
gtgactggagttcagacgtgtgctcttccgatctACCCAGTGTCCACAAG

PD1 Fwd
1632
acactctttccctacacgacgctcttccgatctAGCTTGTCCGTCTGGTT

PD1 Rev
1633
gtgactggagttcagacgtgtgctcttccgatctAGACTCCCCAGACAGG

TGFBR2 Fwd
1634
acactctttccctacacgacgctcttccgatctCATGATAGTCACTGACAACAAC

G

TGFBR2 Rev
1635
gtgactggagttcagacgtgtgctcttccgatctCTTACCATACAGCCACACAGA

TRAC1 Fwd
1636
acactctttccctacacgacgctcttccgatctACATGCAAGCCCATAACC

TRAC1 Rev
1637
gtgactggagttcagacgtgtgctcttccgatctGGTTTGCTCTCAGAGCTTA

B2M-12 Fwd
1638
acactctttccctacacgacgctcttccgatctCATCCGACATTGAAGTTGACTT

A

B2M-12 Rev
1639
gtgactggagttcagacgtgtgctcttccgatctCTACTCATACACAACTTTCAG

CA

CIITA Fwd
1640
acactctttccctacacgacgctcttccgatctTGGGATTCCTACACAATGC

CIITA Rev
1641
gtgactggagttcagacgtgtgctcttccgatctCCCATCCCCAATTCAGAATG

CISH Fwd
1642
acactctttccctacacgacgctcttccgatctCACAAGGCTGACCACATC

CISH Rev
1643
gtgactggagttcagacgtgtgctcttccgatctCAGAAGGCACGTTCTTAGT

TRAC2 Fwd
1644
acactctttccctacacgacgctcttccgatctATCCTCTTGTCCCACAGATATC

TRAC2 Rev
1645
gtgactggagttcagacgtgtgctcttccgatctCCATAGACCTCATGTCTAGCA

Example 7

Another mutation, K516P, was stacked with the best ERCAS12A stacked mutants identified in previous examples (e.g., D303R/S469K/K516P/T826F/F840L). Site directed mutagenesis was used to add this point mutation and the primers are listed in Table 12. The protein was expressed in E. coli BL21 DE3 cells, sequentially purified using nickel-affinity, heparin chromatography and hydroxy-apatite chromatography. To evaluate the genome editing efficiency of each variant, ERCAS12A mutant-RNPs were assembled using crRNAs found in Table 13. The assembled RNPs (2 μM) were delivered into HEK293 cells by nucleofection, and the percent indels was measured by NGS 48-hours post-delivery (FIG. 13). Each of the stacked mutants (D303R/S469K/T826F/F840L; F465R/S469K/T826F/F840L; D303R/S469K/K516P/T826F/F840L) improved activity over ERCAS12A WT at these sites. Stacking these beneficial mutations led to more potent variants of ERCAS12A than any individual point mutations.

TABLE 12

Primers for Site Directed Mutagenesis of ERCAS12A

SEQ ID

Primer
NO
DNA Primers (5′→3′)

K516P Fwd
1654
CTCAGAAGCCGTACTCAACTCCGAAGATCAAGTTAAATTTTGGAATTCCGA

K516P Rev
1655
TCGGAATTCCAAAATTTAACTTGATCTTCGGAGTTGAGTACGGCTTCTGAG

TABLE 13

crRNA for Editing Experiments and NGS Primer Sequences

SEQ ID

Name
NO
crRNA RNA Sequences (5′→3′)

crRNA-1
1656
UUAAUUUCUACUCUUGUAGAUGACCCCUAAGGGAAGAAUGA

crRNA-2
1657
UUAAUUUCUACUCUUGUAGAUCCGAUAUUCCUCAGGUACUC

crRNA-3
1658
UUAAUUUCUACUCUUGUAGAUACUUUCCAUUCUCUGCUGGA

crRNA-4
1659
UUAAUUUCUACUCUUGUAGAUCUGAAUUGCUAUGUGUCUGG

crRNA-5
1660
UUAAUUUCUACUCUUGUAGAUAUCCAUCCGACAUUGAAGUU

crRNA-6
1661
UUAAUUUCUACUCUUGUAGAUUCACAGCCCAAGAUAGUUAA

crRNA-7
1662
UUAAUUUCUACUCUUGUAGAUCCGGCCUUUUUACCUUGGGG

crRNA-8
1663
UUAAUUUCUACUCUUGUAGAUACUGACAGCGUGAACAGGUA

crRNA-9
1664
UUAAUUUCUACUCUUGUAGAUGCACGAAGCUCUCCGAUGUG

crRNA-10
1665
UUAAUUUCUACUCUUGUAGAUUGAUGUGAGAUUUUCCACCU

crRNA-11
1666
UUAAUUUCUACUCUUGUAGAUCAGAUACGAACCUAAACUUU

crRNA-12
1667
UUAAUUUCUACUCUUGUAGAUGAGUCUCUCAGCUGGUACAC

The nucleotide and polypeptide sequences of selected ERCAS12A variants are provided in Table 14.

TABLE 14

Polynucleotide and Polypeptide Sequences of ERCAS12A Variants

ErCas12a-Q18V-NT (SEQ ID NO: 3)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGGTGAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Q18V-AA (SEQ ID NO: 4)

MNNGTNNFQNFIGISSLVKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I35L-NT (SEQ ID NO: 5)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCCTGGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I35L-AA (SEQ ID NO: 6)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-G39Q-NT (SEQ ID NO: 7)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACCAGATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-G39Q-AA (SEQ ID NO: 8)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNQIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-G39K-NT (SEQ ID NO: 9)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACAAAATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-G39K-AA (SEQ ID NO: 10)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNKIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I40Q-NT (SEQ ID NO: 11)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCCAGATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I40Q-AA (SEQ ID NO: 12)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGQIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-D56R-NT (SEQ ID NO: 13)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGCGT

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D56R-AA (SEQ ID NO: 14)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKRIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-D56P-NT (SEQ ID NO: 15)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGCCG

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D56P-AA (SEQ ID NO: 16)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKPIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D59R-NT (SEQ ID NO: 17)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGCGTGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D59R-AA (SEQ ID NO: 18)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMRDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D59A-NT (SEQ ID NO: 19)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGCGGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D59A-AA (SEQ ID NO: 20)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMADYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-S158G-NT (SEQ ID NO: 21)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTGGCCGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S158G-AA (SEQ ID NO: 22)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFGRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDESDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K165Y-NT (SEQ ID NO: 23)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTTATGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K165Y-AA (SEQ ID NO: 24)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFYDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S181V-NT (SEQ ID NO: 25)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTGTGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S181V-AA (SEQ ID NO: 26)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDIVSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-A290K-NT (SEQ ID NO: 27)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCAAAGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-A290K-AA (SEQ ID NO: 28)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIKDTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-D291N-NT (SEQ ID NO: 29)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCACCACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D291N-AA (SEQ ID NO: 30)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIANTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-V296R-NT (SEQ ID NO: 31)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAACGTCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-V296R-AA (SEQ ID NO: 32)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYERPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-P297V-NT (SEQ ID NO: 33)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCGTGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-P297V-AA (SEQ ID NO: 34)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVVYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-P297R-NT (SEQ ID NO: 35)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCGTTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-P297R-AA (SEQ ID NO: 36)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVRYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-P297F-NT (SEQ ID NO: 37)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCTTTTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-P297F-AA (SEQ ID NO: 38)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMESAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVFYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDESDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D303R-NT (SEQ ID NO: 39)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCCGTGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D303R-AA (SEQ ID NO: 40)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMESAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESREEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D303C-NT (SEQ ID NO: 41)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCTGCGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D303C-AA (SEQ ID NO: 42)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESCEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-E305K-NT (SEQ ID NO: 43)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAAAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-E305K-AA (SEQ ID NO: 44)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEKVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y307K-NT (SEQ ID NO: 45)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTAAAACAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y307K-AA (SEQ ID NO: 46)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVKQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Q353R-NT (SEQ ID NO: 47)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCGTAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Q353R-AA (SEQ ID NO: 48)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMESAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSRKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-L444W-NT (SEQ ID NO: 49)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATTGGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-L444W-AA (SEQ ID NO: 50)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHWVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-F465R-NT (SEQ ID NO: 51)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCCGTCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-F465R-AA (SEQ ID NO: 52)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNARHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S469K-NT (SEQ ID NO: 53)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S469K-AA (SEQ ID NO: 54)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-V470K-NT (SEQ ID NO: 55)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCAAATTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-V470K-AA (SEQ ID NO: 56)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSKFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-1498K-NT (SEQ ID NO: 57)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGAAATCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-1498K-AA (SEQ ID NO: 58)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVKSLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYENDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y507W-NT (SEQ ID NO: 59)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTGGGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y507W-AA (SEQ ID NO: 60)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNWVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDESDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-Y507S-NT (SEQ ID NO: 61)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATAGCGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y507S-AA (SEQ ID NO: 62)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNSVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-Y507R-NT (SEQ ID NO: 63)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATCGTGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y507R-AA (SEQ ID NO: 64)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNRVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Q510Y-NT (SEQ ID NO: 65)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTTATAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Q510Y-AA (SEQ ID NO: 66)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLKPYSTKKIKLNFGIPTLADGWSKSKEY

SNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGPNKMIPKVFLSSKTGVETYKPSAYILEGYKQNKH

IKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYREVELQGYKIDWTYISEKDIDLLQEKGQLYLFQIY

NKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNPIIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIP

ENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYFLHMPITINFKANKTGFINDRILQYIAKEKDLHV

IGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIARKEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNA

IIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLLKGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKI

DPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQNTVMSKSSWSVYTYGVRIKRRFVNGRESNESDT

IDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSLSELEDRDYDRLISPVLNENNIFYDSAKAGDALP

KDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNKRYLQKSITEINELVSNYKLCSDDNIKAETYIHE

ISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLVRNY

VTY

ErCas12a-Q510V-NT (SEQ ID NO: 67)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTGTGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Q510V-AA (SEQ ID NO: 68)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTVKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Q510K-NT (SEQ ID NO: 69)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTAAAAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Q510K-AA (SEQ ID NO: 70)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTKKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-P512V-NT (SEQ ID NO: 71)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGGTGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-P512V-AA (SEQ ID NO: 72)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKVYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-P512G-NT (SEQ ID NO: 73)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGGGCTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-P512G-AA (SEQ ID NO: 74)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKGYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y513S-NT (SEQ ID NO: 75)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGAGCTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y513S-AA (SEQ ID NO: 76)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPSSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-Y513R-NT (SEQ ID NO: 77)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGCGTTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y513R-AA (SEQ ID NO: 78)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPRSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y513L-NT (SEQ ID NO: 79)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGCTGTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y513L-AA (SEQ ID NO: 80)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPLSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y513G-NT (SEQ ID NO: 81)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGGGCTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y513G-AA (SEQ ID NO: 82)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPGSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRESNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S514R-NT (SEQ ID NO: 83)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACCGTACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S514R-AA (SEQ ID NO: 84)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYRTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-S514P-NT (SEQ ID NO: 85)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACCCGACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S514P-AA (SEQ ID NO: 86)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYPTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-T515P-NT (SEQ ID NO: 87)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCACCGAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-T515P-AA (SEQ ID NO: 88)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSPKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K516R-NT (SEQ ID NO: 89)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTCGTAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K516R-AA (SEQ ID NO: 90)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTRKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-K516P-NT (SEQ ID NO: 91)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTCCGAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K516P-AA (SEQ ID NO: 92)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTPKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K517P-NT (SEQ ID NO: 93)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAACCGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K517P-AA (SEQ ID NO: 94)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKPIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDESDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I518P-NT (SEQ ID NO: 95)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGCCGAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I518P-AA (SEQ ID NO: 96)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKPKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-L520M-NT (SEQ ID NO: 97)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGATGAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-L520M-AA (SEQ ID NO: 98)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKMNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-F522A-NT (SEQ ID NO: 99)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATGCGGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-F522A-AA (SEQ ID NO: 100)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMESAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNAGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-A541V-NT (SEQ ID NO: 101)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGTGATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-A541V-AA (SEQ ID NO: 102)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNVIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-G553M-NT (SEQ ID NO: 103)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGATGATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-G553M-AA (SEQ ID NO: 104)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLMIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-N556R-NT (SEQ ID NO: 105)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCCGTGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-N556R-AA (SEQ ID NO: 106)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFRAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-N556E-NT (SEQ ID NO: 107)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCGAAGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-N556E-AA (SEQ ID NO: 108)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFEAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-D562V-NT (SEQ ID NO: 109)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGTGAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D562V-AA (SEQ ID NO: 110)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPVKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D562K-NT (SEQ ID NO: 111)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCAAAAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D562K-AA (SEQ ID NO: 112)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPKKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D562I-NT (SEQ ID NO: 113)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCATTAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D562I-AA (SEQ ID NO: 114)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPIKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-N583V-NT (SEQ ID NO: 115)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATGTGCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-N583V-AA (SEQ ID NO: 116)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYVLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-L585K-NT (SEQ ID NO: 117)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTAAACCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-L585K-AA (SEQ ID NO: 118)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLKPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-F653R-NT (SEQ ID NO: 119)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATCGTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-F653R-AA (SEQ ID NO: 120)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNRGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K712R-NT (SEQ ID NO: 121)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAACGTTCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K712R-AA (SEQ ID NO: 122)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKRSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-N716R-NT (SEQ ID NO: 123)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCCGTGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-N716R-AA (SEQ ID NO: 124)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGRDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-E731P-NT (SEQ ID NO: 125)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAACCGAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-E731P-AA (SEQ ID NO: 126)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMESAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEPNLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKEDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-N732T-NT (SEQ ID NO: 127)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAACCTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-N732T-AA (SEQ ID NO: 128)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTS

LFEKMEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSR

FATSFKDYFKNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYE

KYGEFITQEGISFYNDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLD

NISSKHIVERLRKIGDNYNGYNLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQ

KSITEINELVSNYKLCSDDNIKAETYIHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTE

ELVDKDNNFYAELEEIYDEIYPVISLYNLVRNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGI

FNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGPNKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFC

HDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYREVELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKK

STGNDNLHTMYLKNLFSEETLKDIVLKLNGEAEIFFRKSSIKNPIIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIY

QELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYFLHMPITINFKANKTGFINDRILQYIAKEKDL

HVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIARKEWKEIGKIKEIKEGYLSLVIHEISKM

VIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLLKGYQLTYIPDKLKNVGHQCGCI

FYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQNTVMSKSSWSVYTYGV

RIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSLSELEDRDYDR

LISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNKRYL

ErCas12a-A744T-NT (SEQ ID NO: 129)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGACCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-A744T-AA (SEQ ID NO: 130)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGETEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-A744P-NT (SEQ ID NO: 131)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGCCGGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-A744P-AA (SEQ ID NO: 132)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEPEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYENDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-A744N-NT (SEQ ID NO: 133)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGAACGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-A744N-AA (SEQ ID NO: 134)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGENEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-F748A-NT (SEQ ID NO: 135)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTGCGCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-F748A-AA (SEQ ID NO: 136)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFARKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S751A-NT (SEQ ID NO: 137)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAGCGTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S751A-AA (SEQ ID NO: 138)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKASIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSENIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K754G-NT (SEQ ID NO: 139)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTGGCAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K754G-AA (SEQ ID NO: 140)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIGNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-I757W-NT (SEQ ID NO: 141)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

TGGATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I757W-AA (SEQ ID NO: 142)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

WIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I757K-NT (SEQ ID NO: 143)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

AAAATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I757K-AA (SEQ ID NO: 144)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

KIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-E771R-NT (SEQ ID NO: 145)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATCGTGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-E771R-AA (SEQ ID NO: 146)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYRAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-N787A-NT (SEQ ID NO: 147)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGGCGATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-N787A-AA (SEQ ID NO: 148)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKAIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-A825R-NT (SEQ ID NO: 149)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCACGTACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-A825R-AA (SEQ ID NO: 150)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEARTNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-T826K-NT (SEQ ID NO: 151)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAAAAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-T826K-AA (SEQ ID NO: 152)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAKNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-T826F-NT (SEQ ID NO: 153)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-T826F-AA (SEQ ID NO: 154)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y832R-NT (SEQ ID NO: 155)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACCGTCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y832R-AA (SEQ ID NO: 156)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDRRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-Y836Q-NT (SEQ ID NO: 157)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTCAGGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y836Q-AA (SEQ ID NO: 158)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTQDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y836K-NT (SEQ ID NO: 159)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTAAAGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y836K-AA (SEQ ID NO: 160)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTKDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y839L-NT (SEQ ID NO: 161)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGCTGTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y839L-AA (SEQ ID NO: 162)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKLF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-F840L-NT (SEQ ID NO: 163)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACCTG

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-F840L-AA (SEQ ID NO: 164)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y907V-NT (SEQ ID NO: 165)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTGTGGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y907V-AA (SEQ ID NO: 166)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGVDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y907I-NT (SEQ ID NO: 167)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTATTGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y907I-AA (SEQ ID NO: 168)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGIDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-Q916Y-NT (SEQ ID NO: 169)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAATATGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Q916Y-AA (SEQ ID NO: 170)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQYEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-E935R-NT (SEQ ID NO: 171)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGCGTATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-E935R-AA (SEQ ID NO: 172)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKRIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I936R-NT (SEQ ID NO: 173)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAACGTAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I936R-AA (SEQ ID NO: 174)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKERKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K974G-NT (SEQ ID NO: 175)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCGGCGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K974G-AA (SEQ ID NO: 176)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFGVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-K1019V-NT (SEQ ID NO: 177)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATGTGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-K1019V-AA (SEQ ID NO: 178)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDVLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I35L/S469K-NT (SEQ ID NO: 179)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCCTGGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I35L/S469K-AA (SEQ ID NO: 180)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I35L/T826F-NT (SEQ ID NO: 181)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCCTGGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I35L/T826F-AA (SEQ ID NO: 182)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVEMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYENDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-140Q/S469K-NT (SEQ ID NO: 183)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCCAGATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I40Q/S469K-AA (SEQ ID NO: 184)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGQIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-I40Q/T826F-NT (SEQ ID NO: 185)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCCAGATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I40Q/T826F-AA (SEQ ID NO: 186)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGQIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKEDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S469K/K516P-NT (SEQ ID NO: 187)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTCCGAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S469K/K516P-AA (SEQ ID NO: 188)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVEMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTPKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S469K/T826F-NT (SEQ ID NO: 189)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S469K/T826F-AA (SEQ ID NO: 190)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y513G/Y832R-NT (SEQ ID NO: 191)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGGGCTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACCGTCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y513G/Y832R-AA (SEQ ID NO: 192)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPGSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDRRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSENIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-Y513G/F840L-NT (SEQ ID NO: 193)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAGCGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGGGCTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCAACAAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACCTG

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-Y513G/F840L-AA (SEQ ID NO: 194)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCSVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPGSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDESDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAATNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I35L/S469K/T826F-NT (SEQ ID NO: 195)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCCTGGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I35L/S469K/T826F-AA (SEQ ID NO: 196)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-I40Q/S469K/T826F-NT (SEQ ID NO: 197)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCCAGATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-I40Q/S469K/T826F-AA (SEQ ID NO: 198)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGQIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

ErCas12a-S469K/K516P/T826F-NT (SEQ ID NO: 199)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTCCGAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACTTT

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S469K/K516P/T826F-AA (SEQ ID NO: 200)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLESRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTPKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYF

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-S469K/T826F/F840L-NT (SEQ ID NO: 201)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACCTG

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-S469K/T826F/F840L-AA (SEQ ID NO: 202)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D303R/S469K/T826F/F840L-NT (SEQ ID NO: 203)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCCGTGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCTTCCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACCTG

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D303R/S469K/T826F/F840L-AA (SEQ ID NO: 204)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCESADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESREEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-F465R/S469K/T826F/F840L-NT (SEQ ID NO: 205)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCGACGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCCGTCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACCTG

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-F465R/S469K/T826F/F840L-AA (SEQ ID NO: 206)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESDEEVYQSVNGFLDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNARHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D303R/F465R/S469K/T826F/F840L-NT (SEQ ID NO: 207)

atgAATAACGGTACCAATAACTTTCAAAATTTTATCGGGATCTCCTCGTTGCAAAAAACTCTTCGTAATGCTTTAATCCCGACT

GAGACAACACAGCAGTTCATCGTTAAAAACGGCATCATTAAAGAAGATGAGCTGCGCGGCGAGAACCGTCAGATTTTAAAGGAC

ATCATGGACGACTATTACCGTGGGTTCATCTCAGAAACATTAAGCAGCATCGATGATATCGACTGGACCTCACTGTTTGAGAAG

ATGGAGATTCAACTTAAGAATGGAGACAACAAAGACACTTTAATTAAGGAGCAGACGGAATATCGTAAAGCTATCCATAAAAAA

TTTGCAAACGATGACCGTTTCAAAAATATGTTTTCCGCAAAATTGATTAGCGACATTTTACCTGAATTCGTAATCCATAACAAC

AACTATTCCGCATCTGAAAAAGAGGAGAAGACCCAAGTTATCAAACTTTTTTCACGTTTCGCGACATCTTTTAAAGACTACTTT

AAGAATCGTGCGAATTGTTTCAGTGCAGATGATATTTCGTCTAGTTCATGTCACCGTATCGTGAATGACAACGCTGAGATTTTC

TTTTCTAATGCGTTGGTGTATCGCCGCATCGTAAAGTCCTTGTCGAATGATGATATCAACAAAATTAGTGGTGACATGAAAGAC

TCCCTGAAAGAAATGTCATTAGAGGAGATTTACAGCTACGAAAAATACGGAGAGTTCATCACTCAGGAAGGTATCTCCTTTTAC

AACGATATTTGCGGCAAAGTTAATAGTTTCATGAATCTGTATTGTCAGAAGAATAAGGAAAACAAGAATCTGTACAAGTTACAA

AAGCTTCATAAACAGATCTTGTGCATCGCCGACACGTCTTATGAAGTCCCGTATAAATTTGAAAGCCGTGAAGAAGTATATCAG

AGTGTGAATGGATTCTTGGACAATATCTCATCAAAACACATTGTTGAACGTTTGCGCAAAATCGGCGACAACTATAACGGATAC

AACTTGGATAAAATTTATATCGTATCAAAATTTTATGAATCAGTATCCCAAAAGACTTACCGTGATTGGGAGACTATTAATACG

GCGTTAGAAATCCATTACAACAACATTCTGCCGGGTAACGGTAAATCGAAAGCGGACAAAGTGAAGAAGGCGGTTAAGAACGAT

CTTCAGAAGTCTATCACGGAGATTAATGAGCTGGTTTCCAATTACAAGTTGTGCTCGGATGATAACATCAAAGCTGAAACCTAT

ATCCATGAAATTTCTCATATTCTTAATAACTTCGAGGCACAAGAATTAAAATATAACCCAGAAATTCATCTGGTTGAGAGCGAA

CTTAAAGCTTCAGAATTAAAAAATGTTTTGGACGTAATTATGAACGCCCGTCATTGGTGCAAAGTGTTTATGACAGAGGAGTTA

GTAGATAAGGATAACAACTTTTATGCAGAATTGGAAGAGATCTATGATGAGATCTATCCAGTGATTTCACTGTATAACTTAGTC

CGTAATTACGTCACTCAGAAGCCGTACTCAACTAAAAAGATCAAGTTAAATTTTGGAATTCCGACATTGGCGGATGGTTGGAGT

AAGTCTAAGGAATACTCAAATAACGCTATTATCTTAATGCGTGACAACCTGTATTACTTGGGTATTTTCAATGCCAAAAATAAG

CCCGATAAAAAAATCATTGAAGGGAATACTTCAGAAAATAAGGGAGATTACAAAAAAATGATTTATAACCTTTTGCCAGGCCCA

AACAAAATGATCCCCAAGGTGTTTTTGAGTAGTAAGACGGGAGTAGAAACATATAAGCCATCAGCGTACATTCTTGAAGGGTAC

AAACAAAATAAACACATCAAGAGCAGCAAGGACTTTGACATTACATTTTGTCATGACTTAATCGACTACTTTAAAAATTGCATT

GCCATTCACCCTGAGTGGAAAAATTTTGGTTTCGACTTCAGTGATACCTCTACCTATGAAGATATCTCTGGTTTTTATCGTGAG

GTAGAACTTCAGGGTTACAAGATCGATTGGACGTACATTTCCGAGAAAGACATTGATCTTTTACAAGAGAAGGGTCAGCTTTAC

TTATTTCAGATCTACAATAAAGACTTTTCGAAAAAATCCACGGGCAACGACAACCTTCATACAATGTACTTAAAGAACCTGTTC

TCTGAAGAAAACTTGAAAGATATCGTGCTTAAATTGAATGGCGAGGCCGAGATTTTTTTCCGTAAAAGCTCGATTAAGAATCCC

ATCATCCACAAGAAAGGCAGCATTCTGGTAAATCGTACCTATGAAGCGGAGGAAAAGGACCAATTTGGTAATATTCAGATTGTC

CGTAAGAACATCCCTGAAAATATCTATCAAGAATTATATAAATATTTCAACGATAAGTCGGACAAGGAACTTAGCGATGAAGCC

GCGAAGTTGAAAAATGTTGTTGGTCACCACGAAGCAGCATTTAACATTGTGAAAGACTATCGCTACACTTACGATAAGTACCTG

TTACATATGCCTATTACGATTAACTTTAAGGCAAATAAGACAGGTTTTATCAACGACCGTATCCTGCAATATATTGCAAAAGAG

AAGGACCTGCATGTCATTGGGATTGATCGCGGTGAACGCAATCTGATTTACGTGAGTGTTATTGATACCTGCGGTAATATTGTT

GAGCAAAAGTCTTTTAATATCGTTAATGGTTACGACTACCAAATTAAGCTTAAACAACAGGAAGGTGCCCGCCAGATCGCCCGT

AAGGAGTGGAAGGAAATCGGAAAAATTAAGGAAATCAAAGAGGGATATCTGAGCTTGGTTATTCATGAAATCTCAAAGATGGTT

ATTAAGTACAACGCCATCATCGCTATGGAAGACTTATCGTATGGGTTTAAAAAGGGTCGTTTCAAAGTTGAGCGCCAGGTCTAC

CAAAAGTTTGAAACGATGCTTATTAATAAATTAAACTACTTGGTTTTCAAAGATATCTCTATTACCGAGAATGGAGGTTTACTG

AAAGGATACCAGTTAACGTATATCCCCGATAAGCTTAAAAACGTGGGCCATCAGTGTGGCTGTATTTTCTATGTGCCCGCGGCA

TATACATCAAAAATTGACCCGACGACGGGTTTCGTTAATATCTTCAAGTTCAAAGATCTGACAGTTGATGCAAAACGCGAATTC

ATTAAGAAGTTCGATAGTATTCGCTATGACTCCGAGAAAAATTTGTTTTGCTTCACTTTTGACTACAATAACTTTATTACTCAG

AATACAGTCATGAGTAAAAGCTCCTGGTCGGTTTACACGTACGGGGTACGCATCAAACGTCGTTTTGTGAATGGTCGTTTTAGT

AATGAGTCGGATACAATTGATATTACCAAGGATATGGAGAAGACTCTGGAAATGACAGACATCAATTGGCGTGATGGACATGAT

TTGCGTCAGGACATTATCGACTATGAGATTGTGCAGCATATTTTCGAAATTTTTCGCCTGACGGTGCAAATGCGTAATTCCTTA

AGCGAACTGGAAGATCGCGACTATGACCGCTTAATTTCTCCTGTGCTGAATGAGAACAACATCTTCTATGACTCAGCGAAAGCC

GGGGACGCTTTGCCAAAGGATGCAGATGCTAACGGTGCATATTGTATTGCTCTGAAAGGCCTTTACGAGATCAAACAAATTACT

GAAAACTGGAAGGAAGATGGAAAGTTTTCCCGTGATAAATTGAAGATCTCTAATAAGGACTGGTTTGACTTTATCCAAAATAAA

CGTTATTTA

ErCas12a-D303R/F465R/S469K/T826F/F840L-AA (SEQ ID NO: 208)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESREEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNARHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTKKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTFDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWFDFIQNK

RYL

ErCas12a-D303R/S469K/K516P/T826F/F840L-NT (SEQ ID NO: 209)

atgAATAACGGCACCAACAACTTCCAGAACTTTATTGGTATTAGCAGCCTGCAGAAAACCCTGCGTAATGCACTGATTCCGACC

GAAACCACACAGCAGTTTATTGTGAAAAACGGCATCATCAAAGAAGATGAACTTCGCGGAGAAAATCGCCAGATTCTGAAAGAT

ATCATGGATGATTATTATCGCGGTTTTATCAGCGAAACCCTGAGCAGCATTGATGATATTGATTGGACCAGCCTGTTCGAGAAA

ATGGAAATCCAGCTGAAAAATGGCGATAACAAAGACACCCTGATTAAAGAACAGACGGAATATCGTAAAGCCATCCACAAAAAG

TTCGCCAATGATGACCGCTTTAAAAACATGTTCAGCGCAAAACTGATTAGCGATATTCTGCCGGAATTTGTGATCCACAATAAT

AACTATAGCGCCAGCGAGAAAGAAGAGAAAACCCAGGTTATTAAACTGTTTAGCCGTTTTGCGACCAGCTTCAAAGATTACTTC

AAAAATCGTGCCAACTGCTTCAGCGCAGATGATATTAGCAGTAGCAGCTGTCATCGTATTGTGAATGATAACGCCGAAATCTTT

TTCAGCAATGCCCTGGTTTATCGTCGCATTGTTAAAAGCCTGAGCAACGACGACATCAACAAAATCAGCGGTGATATGAAAGAT

AGCCTGAAAGAAATGAGCCTGGAAGAAATCTATAGCTACGAAAAATATGGCGAGTTCATTACCCAAGAAGGCATCAGCTTTTAT

AACGATATTTGCGGCAAAGTGAACAGCTTTATGAATCTGTATTGCCAGAAAAACAAAGAGAACAAAAACCTGTACAAACTGCAG

AAACTGCATAAGCAGATTCTGTGTATTGCAGATACCAGCTATGAAGTGCCGTATAAATTCGAAAGTCGTGAAGAAGTGTACCAG

AGCGTTAATGGTTTTCTGGATAACATCAGCAGCAAACACATTGTTGAACGTCTGCGTAAAATTGGCGACAACTATAATGGCTAT

AACCTGGACAAAATCTACATCGTGAGCAAATTCTATGAAAGCGTGAGCCAGAAAACATATCGTGATTGGGAAACAATTAATACC

GCCTTGGAGATCCACTACAATAACATTCTGCCTGGTAATGGTAAAAGCAAAGCCGACAAAGTTAAAAAGGCCGTGAAAAATGAT

CTGCAGAAATCCATTACCGAAATCAACGAACTGGTCAGCAACTATAAACTGTGTTCCGATGATAATATCAAGGCCGAAACCTAT

ATTCACGAAATCTCCCATATCCTGAACAACTTTGAAGCACAAGAGCTGAAATATAACCCGGAAATTCATCTGGTTGAAAGCGAA

CTGAAAGCCAGTGAACTGAAAAACGTTCTGGATGTGATTATGAACGCCTTTCATTGGTGTAAAGTTTTTATGACCGAAGAACTG

GTGGATAAGGATAACAACTTTTATGCCGAACTGGAAGAAATCTACGATGAAATTTATCCGGTGATCAGCCTGTATAATCTGGTG

CGTAATTATGTGACCCAGAAACCGTATAGCACCCCGAAGATCAAACTGAACTTTGGTATTCCGACACTGGCAGATGGTTGGAGC

AAAAGTAAAGAATATAGCAACAACGCCATTATCCTGATGCGCGATAATCTGTACTATCTGGGTATCTTTAACGCCAAAAACAAA

CCGGACAAGAAAATCATTGAAGGCAATACCAGCGAAAACAAAGGCGACTATAAGAAAATGATTTATAATCTGCTGCCTGGTCCG

AACAAAATGATTCCGAAAGTTTTTCTGAGCAGTAAGACCGGTGTGGAAACATATAAACCGAGCGCATATATTCTGGAAGGCTAC

AAACAGAACAAGCATATCAAAAGCAGCAAAGACTTTGATATCACCTTTTGCCATGATCTGATCGACTATTTCAAAAACTGCATT

GCCATTCATCCGGAATGGAAAAATTTCGGCTTTGATTTCAGCGATACCAGCACCTATGAAGATATTAGCGGTTTTTATCGTGAA

GTGGAACTGCAGGGCTATAAAATCGATTGGACCTATATTAGCGAGAAGGATATTGATCTGCTGCAAGAAAAAGGTCAGCTGTAC

CTGTTTCAGATCTACAACAAAGACTTCAGCAAGAAAAGCACCGGCAATGATAATCTGCATACCATGTATCTGAAGAACCTGTTT

AGCGAAGAGAACCTGAAAGACATTGTGCTGAAACTGAATGGTGAAGCCGAGATTTTCTTTCGCAAATCCAGCATTAAAAACCCG

ATCATCCATAAAAAGGGTAGCATTCTGGTTAATCGTACGTATGAAGCCGAAGAGAAAGATCAGTTTGGCAACATTCAGATTGTG

CGTAAAAACATCCCGGAAAACATCTATCAAGAGCTGTACAAATACTTTAACGATAAAAGCGATAAAGAACTGTCCGATGAAGCA

GCGAAACTGAAGAATGTTGTTGGTCATCATGAAGCCGCATTTAACATTGTGAAAGATTATCGTTACACCTATGATAAATACCTG

CTGCACATGCCGATCACCATCAACTTTAAAGCAAATAAGACGGGTTTCATCAACGATCGCATTCTGCAGTATATCGCCAAAGAA

AAAGACCTGCATGTGATTGGTATTGATCGTGGTGAACGCAATCTGATTTATGTGAGCGTTATTGATACCTGCGGCAATATTGTG

GAACAGAAAAGCTTTAATATCGTGAACGGCTATGACTACCAGATCAAGCTGAAACAGCAAGAAGGTGCACGTCAGATTGCACGT

AAAGAATGGAAAGAAATCGGCAAAATCAAAGAAATTAAAGAAGGCTATCTGAGCCTGGTGATTCATGAGATTAGCAAAATGGTG

ATCAAGTATAACGCCATCATTGCCATGGAAGATCTGAGTTATGGTTTTAAGAAAGGTCGCTTTAAGGTGGAACGTCAGGTGTAT

CAGAAATTTGAAACCATGCTGATCAACAAACTGAATTATCTGGTGTTCAAGGACATCAGCATTACGGAAAATGGTGGTCTGCTG

AAAGGTTATCAGCTGACGTATATTCCGGATAAGCTTAAAAATGTGGGTCATCAGTGTGGCTGCATCTTTTATGTTCCGGCAGCC

TATACCAGCAAAATTGATCCGACCACCGGTTTTGTGAACATCTTCAAATTCAAAGATCTGACCGTTGATGCCAAACGCGAATTC

ATCAAAAAGTTTGATAGCATCCGTTACGACTCCGAGAAAAACCTTTTCTGCTTTACCTTCGACTACAACAACTTTATTACCCAG

AATACGGTGATGAGCAAAAGCAGCTGGTCAGTTTATACCTATGGTGTGCGTATTAAACGTCGCTTTGTGAATGGTCGTTTTAGC

AATGAAAGCGACACCATCGATATCACCAAAGATATGGAAAAGACCCTGGAAATGACCGATATTAACTGGCGTGATGGTCATGAT

CTGCGTCAGGATATTATCGATTATGAAATCGTGCAGCACATCTTTGAAATCTTTCGTCTGACCGTGCAGATGCGTAATAGCCTG

AGCGAGCTGGAAGATCGCGATTATGATCGTCTGATTAGTCCGGTTCTGAACGAAAACAACATCTTCTATGATAGCGCAAAAGCC

GGTGATGCACTGCCGAAAGATGCCGATGCAAATGGTGCATATTGTATTGCACTGAAAGGCCTGTATGAAATCAAGCAGATTACC

GAGAATTGGAAAGAGGATGGTAAATTCAGCCGTGATAAACTGAAAATTAGCAACAAGGATTGGTTCGATTTCATTCAGAACAAA

CGCTACCTG

ErCas12a-D303R/S469K/K516P/T826F/F840L-AA (SEQ ID NO: 210)

MNNGTNNFQNFIGISSLQKTLRNALIPTETTQQFIVKNGIIKEDELRGENRQILKDIMDDYYRGFISETLSSIDDIDWTSLFEK

MEIQLKNGDNKDTLIKEQTEYRKAIHKKFANDDRFKNMFSAKLISDILPEFVIHNNNYSASEKEEKTQVIKLFSRFATSFKDYF

KNRANCFSADDISSSSCHRIVNDNAEIFFSNALVYRRIVKSLSNDDINKISGDMKDSLKEMSLEEIYSYEKYGEFITQEGISFY

NDICGKVNSFMNLYCQKNKENKNLYKLQKLHKQILCIADTSYEVPYKFESREEVYQSVNGELDNISSKHIVERLRKIGDNYNGY

NLDKIYIVSKFYESVSQKTYRDWETINTALEIHYNNILPGNGKSKADKVKKAVKNDLQKSITEINELVSNYKLCSDDNIKAETY

IHEISHILNNFEAQELKYNPEIHLVESELKASELKNVLDVIMNAFHWCKVFMTEELVDKDNNFYAELEEIYDEIYPVISLYNLV

RNYVTQKPYSTPKIKLNFGIPTLADGWSKSKEYSNNAIILMRDNLYYLGIFNAKNKPDKKIIEGNTSENKGDYKKMIYNLLPGP

NKMIPKVFLSSKTGVETYKPSAYILEGYKQNKHIKSSKDFDITFCHDLIDYFKNCIAIHPEWKNFGFDFSDTSTYEDISGFYRE

VELQGYKIDWTYISEKDIDLLQEKGQLYLFQIYNKDFSKKSTGNDNLHTMYLKNLFSEENLKDIVLKLNGEAEIFFRKSSIKNP

IIHKKGSILVNRTYEAEEKDQFGNIQIVRKNIPENIYQELYKYFNDKSDKELSDEAAKLKNVVGHHEAAFNIVKDYRYTYDKYL

LHMPITINFKANKTGFINDRILQYIAKEKDLHVIGIDRGERNLIYVSVIDTCGNIVEQKSFNIVNGYDYQIKLKQQEGARQIAR

KEWKEIGKIKEIKEGYLSLVIHEISKMVIKYNAIIAMEDLSYGFKKGRFKVERQVYQKFETMLINKLNYLVFKDISITENGGLL

KGYQLTYIPDKLKNVGHQCGCIFYVPAAYTSKIDPTTGFVNIFKFKDLTVDAKREFIKKFDSIRYDSEKNLFCFTEDYNNFITQ

NTVMSKSSWSVYTYGVRIKRRFVNGRFSNESDTIDITKDMEKTLEMTDINWRDGHDLRQDIIDYEIVQHIFEIFRLTVQMRNSL

SELEDRDYDRLISPVLNENNIFYDSAKAGDALPKDADANGAYCIALKGLYEIKQITENWKEDGKFSRDKLKISNKDWEDFIQNK

RYL

EUBACTERIUM RECTALE CAS12A MUTANTS

Information

Publication Number

Date Filed

Date Published

Inventors

Original Assignees

CPC

International Classifications

Abstract

Description

Claims

CROSS-REFERENCE TO RELATED APPLICATIONS

Provisional Applications (1)