(Circular) (Six-base) MAPSORT of: pF1KA1793.seq Check: 4278 from: 1 to: 6575 pF1KA1793 6575 bp With 182 enzymes: SgfI * June 20, 2008 10:21 .. AarI CACCTGCnnnn'nnnn_ Cuts at: 2163 2593 2163 Size: 430 6145 AatII G_ACGT'C Cuts at: 943 3565 943 Size: 2622 3953 AccI GT'mk_AC Cuts at: 3613 3613 Size: 6575 Acc65I G'GTAC_C Cuts at: 3591 3591 Size: 6575 AclI AA'CG_TT Cuts at: 6415 6415 Size: 6575 AcuI CTGAAGnnnnnnnnnnnnnn_nn' Cuts at: 944 1046 1228 1276 1534 2657 2800 4400 4832 Size: 102 182 48 258 1123 143 1600 432 Cuts at: 4832 5579 5847 944 Size: 747 268 1672 Fragments arranged by size: 1672 1600 1123 747 432 268 258 182 143 102 48 AfeI AGC'GCT Cuts at: 2650 2650 Size: 6575 AflII C'TTAA_G Cuts at: 514 514 Size: 6575 AflIII A'CryG_T Cuts at: 1405 1607 3927 5031 6008 1405 Size: 202 2320 1104 977 1972 Fragments arranged by size: 2320 1972 1104 977 202 AgeI A'CCGG_T Cuts at: 4963 4963 Size: 6575 AlwNI CAG_nnn'CTG Cuts at: 303 564 792 1220 1380 1443 2169 2609 3313 Size: 261 228 428 160 63 726 440 704 Cuts at: 3313 3880 5447 303 Size: 567 1567 1431 Fragments arranged by size: 1567 1431 726 704 567 440 428 261 228 160 63 ApaI G_GGCC'C Cuts at: 834 834 Size: 6575 ApaLI G'TGCA_C Cuts at: 5345 5345 Size: 6575 ApoI r'AATT_y Cuts at: 3579 5928 6184 3579 Size: 2349 256 3970 Fragments arranged by size: 3970 2349 256 AseI AT'TA_AT Cuts at: 20 20 Size: 6575 AsiSI GCG_AT'CGC Cuts at: 104 104 Size: 6575 AvaI C'yCGr_G Cuts at: 332 3041 3595 332 Size: 2709 554 3312 Fragments arranged by size: 3312 2709 554 BaeI ACnnnnGTAyCnnnnnnn_nnnnn' Cuts at: 2387 2420 2387 Size: 33 6542 BamHI G'GATC_C Cuts at: 3600 3600 Size: 6575 BanI G'GyrC_C Cuts at: 3591 4005 4253 4288 3591 Size: 414 248 35 5878 Fragments arranged by size: 5878 414 248 35 BanII G_rGCy'C Cuts at: 297 383 730 834 1142 1271 1304 1664 1808 Size: 86 347 104 308 129 33 360 144 Cuts at: 1808 2210 2999 3589 4619 297 Size: 402 789 590 1030 2253 Fragments arranged by size: 2253 1030 789 590 402 360 347 308 144 129 104 86 33 BbeI G_GCGC'C Cuts at: 4009 4257 4009 Size: 248 6327 BbsI GAAGACnn'nnnn_ Cuts at: 662 1940 2645 3389 662 Size: 1278 705 744 3848 Fragments arranged by size: 3848 1278 744 705 BcgI CGAnnnnnnTGCnnnnnnnnnn_nn' Cuts at: 90 124 2608 2642 90 Size: 34 2484 34 4023 Fragments arranged by size: 4023 2484 34 34 BciVI GTATCCnnnnn_n' Cuts at: 1485 3114 4473 5234 6575 1485 Size: 1629 1359 761 1341 1485 Fragments arranged by size: 1629 1485 1359 1341 761 BclI T'GATC_A Cuts at: 2678 4095 2678 Size: 1417 5158 BfrBI ATG'CAT Cuts at: 1038 1038 Size: 6575 BglI GCCn_nnn'nGGC Cuts at: 415 2194 2500 415 Size: 1779 306 4490 Fragments arranged by size: 4490 1779 306 BglII A'GATC_T Cuts at: 873 4090 6127 873 Size: 3217 2037 1321 Fragments arranged by size: 3217 2037 1321 BlpI GC'TnA_GC Cuts at: 1544 2630 3299 3687 1544 Size: 1086 669 388 4432 Fragments arranged by size: 4432 1086 669 388 Bme1580I G_kGCm'C Cuts at: 834 4200 4293 5349 834 Size: 3366 93 1056 2060 Fragments arranged by size: 3366 2060 1056 93 BmrI ACTGGGnnnn_n' Cuts at: 3787 3868 4203 6347 3787 Size: 81 335 2144 4015 Fragments arranged by size: 4015 2144 335 81 BmtI G_CTAG'C Cuts at: 403 2924 403 Size: 2521 4054 BplI GAGnnnnnCTCnnnnnnnn_nnnnn' Cuts at: 378 714 4942 378 Size: 336 4228 2011 Fragments arranged by size: 4228 2011 336 BpmI CTGGAGnnnnnnnnnnnnnn_nn' Cuts at: 301 343 406 898 1372 1612 1642 1669 1831 Size: 42 63 492 474 240 30 27 162 Cuts at: 1831 2986 3178 301 Size: 1155 192 3698 Fragments arranged by size: 3698 1155 492 474 240 192 162 63 42 30 27 Bpu10I CC'TnA_GC Cuts at: 296 3007 296 Size: 2711 3864 BpuEI CTTGAGnnnnnnnnnnnnnn_nn' Cuts at: 1126 1648 1888 2350 2452 3160 3751 4630 5122 Size: 522 240 462 102 708 591 879 492 Cuts at: 5122 5420 1126 Size: 298 2281 Fragments arranged by size: 2281 879 708 591 522 492 462 298 240 102 BsaI GGTCTCn'nnnn_ Cuts at: 322 757 3136 6279 322 Size: 435 2379 3143 618 Fragments arranged by size: 3143 2379 618 435 BsaAI yAC'GTr Cuts at: 3928 3928 Size: 6575 BsaBI GATnn'nnATC Cuts at: 6 691 4089 6126 6 Size: 685 3398 2037 455 Fragments arranged by size: 3398 2037 685 455 BsaHI Gr'CG_yC Cuts at: 940 2186 3407 3562 4006 4254 4956 6452 940 Size: 1246 1221 155 444 248 702 1496 1063 Fragments arranged by size: 1496 1246 1221 1063 702 444 248 155 BsaWI w'CCGG_w Cuts at: 739 1683 3764 4285 4963 5237 5384 5795 739 Size: 944 2081 521 678 274 147 411 1519 Fragments arranged by size: 2081 1519 944 678 521 411 274 147 BsaXI ACnnnnnCTCCnnnnnnn_nnn' Cuts at: 365 395 1129 1159 1195 1225 1348 1378 1444 Size: 30 734 30 36 30 123 30 66 Cuts at: 1444 1474 365 Size: 30 5466 Fragments arranged by size: 5466 734 123 66 36 30 30 30 30 30 BseRI GAGGAGnnnnnnnn_nn' Cuts at: 166 187 298 340 490 1126 1129 1216 1465 Size: 21 111 42 150 636 3 87 249 Cuts at: 1465 1836 1849 2254 3007 3229 3244 3253 3385 Size: 371 13 405 753 222 15 9 132 Cuts at: 3385 3409 3412 166 Size: 24 3 3329 Fragments arranged by size: 3329 753 636 405 371 249 222 150 132 111 87 42 24 21 15 13 9 3 3 BseYI C'CCAG_C Cuts at: 138 410 2365 4000 4758 5335 5831 138 Size: 272 1955 1635 758 577 496 882 Fragments arranged by size: 1955 1635 882 758 577 496 272 BsgI GTGCAGnnnnnnnnnnnnnn_nn' Cuts at: 229 728 1366 1846 2505 2595 2716 2974 229 Size: 499 638 480 659 90 121 258 3830 Fragments arranged by size: 3830 659 638 499 480 258 121 90 BsiEI CG_ry'CG Cuts at: 104 4117 4163 5371 104 Size: 4013 46 1208 1308 Fragments arranged by size: 4013 1308 1208 46 BsiHKAI G_wGCw'C Cuts at: 730 1142 1152 1271 1304 1664 1808 2210 2999 Size: 412 10 119 33 360 144 402 789 Cuts at: 2999 3589 4367 4557 5349 730 Size: 590 778 190 792 1956 Fragments arranged by size: 1956 792 789 778 590 412 402 360 190 144 119 33 10 BsmI GAATG_Cn' Cuts at: 668 2833 3057 3081 668 Size: 2165 224 24 4162 Fragments arranged by size: 4162 2165 224 24 BsmBI CGTCTCn'nnnn_ Cuts at: 1528 3553 1528 Size: 2025 4550 Bsp1286I G_dGCh'C Cuts at: 297 383 730 834 1142 1152 1271 1304 1664 Size: 86 347 104 308 10 119 33 360 Cuts at: 1664 1808 2210 2999 3589 4200 4293 4367 4557 Size: 144 402 789 590 611 93 74 190 Cuts at: 4557 4619 5349 297 Size: 62 730 1523 Fragments arranged by size: 1523 789 730 611 590 402 360 347 308 190 144 119 104 93 86 74 62 33 10 BspEI T'CCGG_A Cuts at: 1683 5795 1683 Size: 4112 2463 BspHI T'CATG_A Cuts at: 1319 5751 1319 Size: 4432 2143 BspMI ACCTGCnnnn'nnnn_ Cuts at: 2163 2593 2634 3625 3839 4141 4522 2163 Size: 430 41 991 214 302 381 4216 Fragments arranged by size: 4216 991 430 381 302 214 41 BsrBI CCG'CTC Cuts at: 50 1596 2244 4867 4921 6405 6570 50 Size: 1546 648 2623 54 1484 165 55 Fragments arranged by size: 2623 1546 1484 648 165 55 54 BsrDI GCAATG_nn' Cuts at: 1524 2376 4487 1524 Size: 852 2111 3612 Fragments arranged by size: 3612 2111 852 BsrFI r'CCGG_y Cuts at: 1367 2188 4573 4754 4963 1367 Size: 821 2385 181 209 2979 Fragments arranged by size: 2979 2385 821 209 181 BssSI C'ACGA_G Cuts at: 4846 5204 4846 Size: 358 6217 BstAPI GCAn_nnn'nTGC Cuts at: 792 1380 1443 2169 2694 2828 3952 792 Size: 588 63 726 525 134 1124 3415 Fragments arranged by size: 3415 1124 726 588 525 134 63 BstBI TT'CG_AA Cuts at: 4936 4936 Size: 6575 BstEII G'GTnAC_C Cuts at: 3102 3102 Size: 6575 BstYI r'GATC_y Cuts at: 1 437 554 627 873 2030 3309 3600 4067 Size: 436 117 73 246 1157 1279 291 467 Cuts at: 4067 4090 4425 4671 5683 5769 5781 6127 1 Size: 23 335 246 1012 86 12 346 449 Fragments arranged by size: 1279 1157 1012 467 449 436 346 335 291 246 246 117 86 73 23 12 Bsu36I CC'TnA_GG Cuts at: 1486 3314 1486 Size: 1828 4747 BtgI C'CryG_G Cuts at: 2843 2843 Size: 6575 BtgZI GCGATGnnnnnnnnnn'nnnn_ Cuts at: 1081 4525 4706 4869 1081 Size: 3444 181 163 2787 Fragments arranged by size: 3444 2787 181 163 BtsI GCAGTG_nn' Cuts at: 228 1465 1720 1804 1873 1879 2986 3283 3818 Size: 1237 255 84 69 6 1107 297 535 Cuts at: 3818 228 Size: 2985 Fragments arranged by size: 2985 1237 1107 535 297 255 84 69 6 DraI TTT'AAA Cuts at: 3574 3574 Size: 6575 DrdI GACnn_nn'nnGTC Cuts at: 4281 5139 4281 Size: 858 5717 EaeI y'GGCC_r Cuts at: 2762 4160 4334 4725 4752 6097 2762 Size: 1398 174 391 27 1345 3240 Fragments arranged by size: 3240 1398 1345 391 174 27 EagI C'GGCC_G Cuts at: 4160 4160 Size: 6575 EarI CTCTTCn'nnn_ Cuts at: 152 158 164 176 718 1022 1076 1850 2033 Size: 6 6 12 542 304 54 774 183 Cuts at: 2033 2242 2338 2506 2548 2587 2780 3010 3347 Size: 209 96 168 42 39 193 230 337 Cuts at: 3347 3362 3386 3420 4598 4808 152 Size: 15 24 34 1178 210 1919 Fragments arranged by size: 1919 1178 774 542 337 304 230 210 209 193 183 168 96 54 42 39 34 24 15 12 6 6 EciI GGCGGAnnnnnnnnn_nn' Cuts at: 2989 4783 5093 5239 6385 2989 Size: 1794 310 146 1146 3179 Fragments arranged by size: 3179 1794 1146 310 146 EcoICRI GAG'CTC Cuts at: 728 1140 1269 1302 1662 1806 2208 2997 3587 Size: 412 129 33 360 144 402 789 590 Cuts at: 3587 728 Size: 3716 Fragments arranged by size: 3716 789 590 412 402 360 144 129 33 Eco57MI CTGrAGnnnnnnnnnnnnnn_nn' Cuts at: 301 343 406 898 944 1046 1228 1276 1372 Size: 42 63 492 46 102 182 48 96 Cuts at: 1372 1534 1612 1642 1669 1831 2657 2800 2986 Size: 162 78 30 27 162 826 143 186 Cuts at: 2986 3178 4400 4832 5579 5847 301 Size: 192 1222 432 747 268 1029 Fragments arranged by size: 1222 1029 826 747 492 432 268 192 186 182 162 162 143 102 96 78 63 48 46 42 30 27 EcoNI CCTnn'n_nnAGG Cuts at: 622 817 2802 3512 622 Size: 195 1985 710 3685 Fragments arranged by size: 3685 1985 710 195 EcoO109I rG'GnC_Cy Cuts at: 831 1629 2166 3714 831 Size: 798 537 1548 3692 Fragments arranged by size: 3692 1548 798 537 EcoRI G'AATT_C Cuts at: 3579 3579 Size: 6575 FalI AAGnnnnnCTTnnnnnnnn_nnnnn' Cuts at: 4594 4594 Size: 6575 FspI TGC'GCA Cuts at: 4356 4356 Size: 6575 HaeII r_GCGC'y Cuts at: 410 2652 4009 4257 5279 6258 410 Size: 2242 1357 248 1022 979 727 Fragments arranged by size: 2242 1357 1022 979 727 248 Hin4I GAynnnnnvTCnnnnnnnn_nnnnn' Cuts at: 248 280 546 578 598 630 1195 1227 1527 Size: 32 266 32 20 32 565 32 300 Cuts at: 1527 1559 3397 3429 3550 3582 4417 4447 4449 Size: 32 1838 32 121 32 835 30 2 Cuts at: 4449 4479 248 Size: 30 2344 Fragments arranged by size: 2344 1838 835 565 300 266 121 32 32 32 32 32 32 32 30 30 20 2 HincII GTy'rAC Cuts at: 243 3614 243 Size: 3371 3204 KasI G'GCGC_C Cuts at: 4005 4253 4005 Size: 248 6327 KpnI G_GTAC'C Cuts at: 3595 3595 Size: 6575 MluI A'CGCG_T Cuts at: 6008 6008 Size: 6575 MmeI TCCrACnnnnnnnnnnnnnnnnnn_nn' Cuts at: 1058 2060 5246 5430 1058 Size: 1002 3186 184 2203 Fragments arranged by size: 3186 2203 1002 184 MscI TGG'CCA Cuts at: 2764 4336 2764 Size: 1572 5003 MslI CAynn'nnrTG Cuts at: 356 602 1610 4564 4691 356 Size: 246 1008 2954 127 2240 Fragments arranged by size: 2954 2240 1008 246 127 MspA1I CmG'CkG Cuts at: 309 1284 1443 1566 1620 1647 1785 1848 1953 Size: 975 159 123 54 27 138 63 105 Cuts at: 1953 2292 2609 2736 2907 2964 2976 3686 4000 Size: 339 317 127 171 57 12 710 314 Cuts at: 4000 4360 5373 5618 309 Size: 360 1013 245 1266 Fragments arranged by size: 1266 1013 975 710 360 339 317 314 245 171 159 138 127 123 105 63 57 54 27 12 NaeI GCC'GGC Cuts at: 1369 4756 1369 Size: 3387 3188 NarI GG'CG_CC Cuts at: 4006 4254 4006 Size: 248 6327 NcoI C'CATG_G Cuts at: 2843 2843 Size: 6575 NgoMIV G'CCGG_C Cuts at: 1367 4754 1367 Size: 3387 3188 NheI G'CTAG_C Cuts at: 399 2920 399 Size: 2521 4054 NsiI A_TGCA'T Cuts at: 1040 1040 Size: 6575 NspI r_CATG'y Cuts at: 1409 1611 1971 3628 4128 5035 1409 Size: 202 360 1657 500 907 2949 Fragments arranged by size: 2949 1657 907 500 360 202 PciI A'CATG_T Cuts at: 1405 1607 5031 1405 Size: 202 3424 2949 Fragments arranged by size: 3424 2949 202 PflMI CCAn_nnn'nTGG Cuts at: 1891 1891 Size: 6575 PfoI T'CCnGG_A Cuts at: 383 1808 2069 383 Size: 1425 261 4889 Fragments arranged by size: 4889 1425 261 PmeI GTTT'AAAC Cuts at: 3574 3574 Size: 6575 PpiI GAACnnnnnCTCnnnnnnnn_nnnnn' Cuts at: 5714 5746 5714 Size: 32 6543 PpuMI rG'GwC_Cy Cuts at: 1629 1629 Size: 6575 PshAI GACnn'nnGTC Cuts at: 2189 2189 Size: 6575 PspOMI G'GGCC_C Cuts at: 830 830 Size: 6575 PstI C_TGCA'G Cuts at: 308 623 788 839 920 1133 1232 1376 1448 Size: 315 165 51 81 213 99 144 72 Cuts at: 1448 1460 1481 1790 1853 1952 2174 2258 2690 Size: 12 21 309 63 99 222 84 432 Cuts at: 2690 2885 2906 2981 3622 4307 308 Size: 195 21 75 641 685 2576 Fragments arranged by size: 2576 685 641 432 315 309 222 213 195 165 144 99 99 84 81 75 72 63 51 21 21 12 PvuI CG_AT'CG Cuts at: 104 104 Size: 6575 PvuII CAG'CTG Cuts at: 309 1284 1443 1566 1620 1647 1785 1848 1953 Size: 975 159 123 54 27 138 63 105 Cuts at: 1953 2292 2609 2736 2907 2964 2976 4000 4360 Size: 339 317 127 171 57 12 1024 360 Cuts at: 4360 309 Size: 2524 Fragments arranged by size: 2524 1024 975 360 339 317 171 159 138 127 123 105 63 57 54 27 12 RsrII CG'GwC_CG Cuts at: 4770 4770 Size: 6575 SacI G_AGCT'C Cuts at: 730 1142 1271 1304 1664 1808 2210 2999 3589 Size: 412 129 33 360 144 402 789 590 Cuts at: 3589 730 Size: 3716 Fragments arranged by size: 3716 789 590 412 402 360 144 129 33 SalI G'TCGA_C Cuts at: 3612 3612 Size: 6575 SapI GCTCTTCn'nnn_ Cuts at: 718 1022 1076 1850 2033 2338 2780 4598 4808 Size: 304 54 774 183 305 442 1818 210 Cuts at: 4808 718 Size: 2485 Fragments arranged by size: 2485 1818 774 442 305 304 210 183 54 SbfI CC_TGCA'GG Cuts at: 623 839 2174 3622 623 Size: 216 1335 1448 3576 Fragments arranged by size: 3576 1448 1335 216 ScaI AGT'ACT Cuts at: 6092 6092 Size: 6575 SfcI C'TryA_G Cuts at: 33 304 373 619 784 835 916 1087 1129 Size: 271 69 246 165 51 81 171 42 Cuts at: 1129 1228 1372 1444 1456 1477 1786 1849 1948 Size: 99 144 72 12 21 309 63 99 Cuts at: 1948 2170 2254 2686 2881 2902 2977 3618 4303 Size: 222 84 432 195 21 75 641 685 Cuts at: 4303 5296 5487 33 Size: 993 191 1121 Fragments arranged by size: 1121 993 685 641 432 309 271 246 222 195 191 171 165 144 99 99 84 81 75 72 69 63 51 42 21 21 12 SfoI GGC'GCC Cuts at: 4007 4255 4007 Size: 248 6327 SgfI GCG_AT'CGC Cuts at: 104 104 Size: 6575 SmaI CCC'GGG Cuts at: 334 3043 3597 334 Size: 2709 554 3312 Fragments arranged by size: 3312 2709 554 SmlI C'TyrA_G Cuts at: 514 1141 1663 1903 2329 2467 3139 3730 4645 Size: 627 522 240 426 138 672 591 915 Cuts at: 4645 5137 5399 514 Size: 492 262 1690 Fragments arranged by size: 1690 915 672 627 591 522 492 426 262 240 138 SpeI A'CTAG_T Cuts at: 67 67 Size: 6575 SphI G_CATG'C Cuts at: 1971 3628 4128 1971 Size: 1657 500 4418 Fragments arranged by size: 4418 1657 500 StuI AGG'CCT Cuts at: 1669 2503 3021 3518 1669 Size: 834 518 497 4726 Fragments arranged by size: 4726 834 518 497 StyI C'CwwG_G Cuts at: 365 450 1962 2843 3709 4063 365 Size: 85 1512 881 866 354 2877 Fragments arranged by size: 2877 1512 881 866 354 85 TaqII GACCGAnnnnnnnnn_nn' Cuts at: 1207 2458 4158 4291 4750 4959 1207 Size: 1251 1700 133 459 209 2823 Fragments arranged by size: 2823 1700 1251 459 209 133 TatI w'GTAC_w Cuts at: 1754 3170 6090 1754 Size: 1416 2920 2239 Fragments arranged by size: 2920 2239 1416 TstI CACnnnnnnTCCnnnnnnn_nnnnn' Cuts at: 1129 1161 1348 1380 1444 1476 6092 6124 1129 Size: 32 187 32 64 32 4616 32 1580 Fragments arranged by size: 4616 1580 187 64 32 32 32 32 Tth111I GACn'n_nGTC Cuts at: 3564 4372 3564 Size: 808 5767 XbaI T'CTAG_A Cuts at: 3606 3606 Size: 6575 XcmI CCAnnnn_n'nnnnTGG Cuts at: 114 2148 3153 4070 114 Size: 2034 1005 917 2619 Fragments arranged by size: 2619 2034 1005 917 XmaI C'CCGG_G Cuts at: 332 3041 3595 332 Size: 2709 554 3312 Fragments arranged by size: 3312 2709 554 XmnI GAAnn'nnTTC Cuts at: 668 668 Size: 6575 ZraI GAC'GTC Cuts at: 941 3563 941 Size: 2622 3953 Enzymes that do cut: AarI AatII AccI Acc65I AclI AcuI AfeI AflII AflIII AgeI AlwNI ApaI ApaLI ApoI AseI AsiSI AvaI BaeI BamHI BanI BanII BbeI BbsI BcgI BciVI BclI BfrBI BglI BglII BlpI Bme1580I BmrI BmtI BplI BpmI Bpu10I BpuEI BsaI BsaAI BsaBI BsaHI BsaWI BsaXI BseRI BseYI BsgI BsiEI BsiHKAI BsmI BsmBI Bsp1286I BspEI BspHI BspMI BsrBI BsrDI BsrFI BssSI BstAPI BstBI BstEII BstYI Bsu36I BtgI BtgZI BtsI DraI DrdI EaeI EagI EarI EciI EcoICRI Eco57MI EcoNI EcoO109I EcoRI FalI FspI HaeII Hin4I HincII KasI KpnI MluI MmeI MscI MslI MspA1I NaeI NarI NcoI NgoMIV NheI NsiI NspI PciI PflMI PfoI PmeI PpiI PpuMI PshAI PspOMI PstI PvuI PvuII RsrII SacI SalI SapI SbfI ScaI SfcI SfoI SgfI SmaI SmlI SpeI SphI StuI StyI TaqII TatI TstI Tth111I XbaI XcmI XmaI XmnI ZraI Enzymes that do not cut: AhdI AleI AlfI AloI AscI AvrII BbvCI BmgBI BsiWI BsrGI BssHII BstXI BstZ17I ClaI CspCI DraIII EcoRV FseI FspAI HindIII HpaI MfeI NdeI NotI NruI PacI PasI PmlI PsiI PspXI PsrI SacII SanDI SexAI SfiI SgrAI SnaBI SrfI SspI SwaI XhoI