Addgene p(bhsp68-Cas9) - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: p(bhsp68-Cas9)


Map View

lac operator M13/pUC Reverse (8279 .. 8301) CAP binding site BspQI - SapI (8085) L4440 (8066 .. 8083) pBR322ori-F (7813 .. 7832) AlwNI (7559) AhdI (7080) NmeAIII (6933) Amp-R (6511 .. 6530) pBRforEco (6102 .. 6120) pGEX 3' (6042 .. 6064) PfoI (6041) pRS-marker (5923 .. 5942) F1ori-F (5748 .. 5769) DraIII (5684) BtgZI (5676) F1ori-R (5538 .. 5557) M13 Forward (5222 .. 5239) XcmI (5219) AflII (5187) SfiI (5175) SnaBI (4927) BsrGI (4852) SacII (4846) BsiWI (4831) NotI (4801) M13 Reverse (8298 .. 8314) Bsu36I (51) PspOMI (69) ApaI (73) BglII (109) XbaI (611) AgeI (632) BstZ17I (859) BmgBI (864) SwaI (1165) BsgI (1267) BstAPI (1425) BfuAI - BspMI (1713) NheI (1743) BmtI (1747) AleI (1891) SphI (1905) BsmI (2248) Acc65I (2418) KpnI (2422) p(bhsp68-Cas9) 8323 bp

Sequence View

Bsu36I 5ʹ 3ʹ 60 g a a t t g g c g c g c c t a g g c c g g c c g a a t t c g a a t g g c c a t g g g a c g t c g a c c t g a g g t a a t c t t a a c c g c g c g g a t c c g g c c g g c t t a a g c t t a c c g g t a c c c t g c a g c t g g a c t c c a t t a

BglIIPspOMIApaI 120 t a t a a c c c g g g c c c t a t a t a t g g a t c c a a t t g c a a t g a t c a t c a t g a c a g a t c t g c g c g c a t a t t g g g c c c g g g a t a t a t a c c t a g g t t a a c g t t a c t a g t a g t a c t g t c t a g a c g c g c g

180 g a t c g a t a t c c g t t t c a t a t a t a a g c g c g g t c t c g c g g c g c g t t g t c a g t g a a t t t g a a t c t a g c t a t a g g c a a a g t a t a t a t t c g c g c c a g a g c g c c g c g c a a c a g t c a c t t a a a c t t a

240 c a a g c g a a g c g a a a g c a a c a a a g c t a a g c a a g c g a c t c a a a g c a a a t t t t c a a a g c g a c t g t t c g c t t c g c t t t c g t t g t t t c g a t t c g t t c g c t g a g t t t c g t t t a a a a g t t t c g c t g a

300 t t a a c a a a g t t g a g t g a a t t a t t t a c t a g t g a a t t c a a a g t c g t c g a t a a a t a a g t t t c c a a t t g t t t c a a c t c a c t t a a t a a a t g a t c a c t t a a g t t t c a g c a g c t a t t t a t t c a a a g g

360 c a t t t t t a t t c c c c a g c c a g g a a g t t a g t t t c a a t a g t t t t g t a a t t t c a a c g a a a c t c a g t a a a a a t a a g g g g t c g g t c c t t c a a t c a a a g t t a t c a a a a c a t t a a a g t t g c t t t g a g t

420 M t t t g a t t t c g t a c t a a t t t t c c a c a t c t c t a t t t t c t g c c c g c a g a a t a a t c c a a a a t g c a a a c t a a a g c a t g a t t a a a a g g t g t a g a g a t a a a a g a c g g g c g t c t t a t t a g g t t t t a c g

480 130 PKKKRKV (in frame with SV40 NLS) EDDPVV SV40 NLS QIGDPTPPKKKRKVEDDPVV a g a t c g g g g a t c c c a c c c c a c c c a a g a a g a a g c g c a a g g t g g a g g a c g a t c c c g t c g t t t t c t a g c c c c t a g g g t g g g g t g g g t t c t t c t t c g c g t t c c a c c t c c t g c t a g g g c a g c a a a

540 (in frame with SV40 NLS) LQRRDWENPGVTQLNRLAAH LQRRDWENPGVTQLNRLAAH t a c a a c g t c g t g a c t g g g a a a a c c c t g g c g t t a c c c a a c t t a a t c g c c t t g c a g c a c a t c a t g t t g c a g c a c t g a c c c t t t t g g g a c c g c a a t g g g t t g a a t t a g c g g a a c g t c g t g t a g

600 (in frame with SV40 NLS) PPFASWRNSEEARTDRPSQQ PPFASWRNSEEARTDRPSQQ c c c c t t t c g c c a g c t g g c g t a a t a g c g a a g a g g c c c g c a c c g a t c g c c c t t c c c a a c a g t g g g g a a a g c g g t c g a c c g c a t t a t c g c t t c t c c g g g c g t g g c t a g c g g g a a g g g t t g t c a

AgeIXbaI 660 (in frame with SV40 NLS) LRSTLEDPRDPPVAT 1 5 MDKKY Cas9 1 5 MDKKY Cas9 Kozak sequence LRSTLEDPRDPPVATMDKKY t g c g g t c g a c t c t a g a g g a t c c c c g g g a t c c a c c g g t c g c c a c c a t g g a t a a g a a a t a c t a c g c c a g c t g a g a t c t c c t a g g g g c c c t a g g t g g c c a g c g g t g g t a c c t a t t c t t t a t g a

720 10 15 20 25 SIGLDIGTNSVGWAVITDEY Cas9 10 15 20 25 SIGLDIGTNSVGWAVITDEY Cas9 SIGLDIGTNSVGWAVITDEY c a a t a g g c t t a g a t a t c g g c a c a a a t a g c g t c g g a t g g g c g g t g a t c a c t g a t g a a t a t a g t t a t c c g a a t c t a t a g c c g t g t t t a t c g c a g c c t a c c c g c c a c t a g t g a c t a c t t a t a t

780 30 35 40 45 KVPSKKFKVLGNTDRHSIKK Cas9 30 35 40 45 KVPSKKFKVLGNTDRHSIKK Cas9 KVPSKKFKVLGNTDRHSIKK a g g t t c c g t c t a a a a a g t t c a a g g t t c t g g g a a a t a c a g a c c g c c a c a g t a t c a a a a a a a t c c a a g g c a g a t t t t t c a a g t t c c a a g a c c c t t t a t g t c t g g c g g t g t c a t a g t t t t t t t

840 50 55 60 65 NLIGALLFDSGETAEATRLK Cas9 50 55 60 65 NLIGALLFDSGETAEATRLK Cas9 NLIGALLFDSGETAEATRLK a t c t t a t a g g g g c t c t t t t a t t t g a c a g t g g a g a g a c a g c g g a a g c g a c t c g t c t c a a a c t a g a a t a t c c c c g a g a a a a t a a a c t g t c a c c t c t c t g t c g c c t t c g c t g a g c a g a g t t t g

BstZ17IBmgBI 900 70 75 80 85 RTARRRYTRRKNRICYLQEI Cas9 70 75 80 85 RTARRRYTRRKNRICYLQEI Cas9 RTARRRYTRRKNRICYLQEI g g a c a g c t c g t a g a a g g t a t a c a c g t c g g a a g a a t c g t a t t t g t t a t c t a c a g g a g a t t t c c t g t c g a g c a t c t t c c a t a t g t g c a g c c t t c t t a g c a t a a a c a a t a g a t g t c c t c t a a a

960 90 95 100 105 FSNEMAKVDDSFFHRLEESF Cas9 90 95 100 105 FSNEMAKVDDSFFHRLEESF Cas9 FSNEMAKVDDSFFHRLEESF t t t c a a a t g a g a t g g c g a a a g t a g a t g a t a g t t t c t t t c a t c g a c t t g a a g a g t c t t t t t a a a g t t t a c t c t a c c g c t t t c a t c t a c t a t c a a a g a a a g t a g c t g a a c t t c t c a g a a a a a

1020 110 115 120 125 LVEEDKKHERHPIFGNIVDE Cas9 110 115 120 125 LVEEDKKHERHPIFGNIVDE Cas9 LVEEDKKHERHPIFGNIVDE t g g t g g a a g a a g a c a a g a a g c a t g a a c g t c a t c c t a t t t t t g g a a a t a t a g t a g a t g a a g a c c a c c t t c t t c t g t t c t t c g t a c t t g c a g t a g g a t a a a a a c c t t t a t a t c a t c t a c t t c

1080 130 135 140 145 VAYHEKYPTIYHLRKKLVDS Cas9 130 135 140 145 VAYHEKYPTIYHLRKKLVDS Cas9 VAYHEKYPTIYHLRKKLVDS t t g c t t a t c a t g a g a a a t a t c c a a c t a t c t a t c a t c t g c g a a a a a a a t t g g t a g a t t c t a a a c g a a t a g t a c t c t t t a t a g g t t g a t a g a t a g t a g a c g c t t t t t t t a a c c a t c t a a g a t

1140 150 155 160 165 TDKADLRLIYLALAHMIKFR Cas9 150 155 160 165 TDKADLRLIYLALAHMIKFR Cas9 TDKADLRLIYLALAHMIKFR c t g a t a a a g c g g a t t t g c g c t t a a t c t a t t t g g c c t t a g c g c a t a t g a t t a a g t t t c g t g g a c t a t t t c g c c t a a a c g c g a a t t a g a t a a a c c g g a a t c g c g t a t a c t a a t t c a a a g c a c

SwaI 1200 170 175 180 185 GHFLIEGDLNPDNSDVDKLF Cas9 170 175 180 185 GHFLIEGDLNPDNSDVDKLF Cas9 GHFLIEGDLNPDNSDVDKLF g t c a t t t t t t g a t t g a g g g a g a t t t a a a t c c t g a t a a t a g t g a t g t g g a c a a a c t a t t t a c a g t a a a a a a c t a a c t c c c t c t a a a t t t a g g a c t a t t a t c a c t a c a c c t g t t t g a t a a a t

1260 190 195 200 205 IQLVQTYNQLFEENPINASG Cas9 190 195 200 205 IQLVQTYNQLFEENPINASG Cas9 IQLVQTYNQLFEENPINASG t c c a g t t g g t a c a a a c c t a c a a t c a a t t a t t t g a a g a a a a c c c t a t t a a c g c a a g t g g a g a g g t c a a c c a t g t t t g g a t g t t a g t t a a t a a a c t t c t t t t g g g a t a a t t g c g t t c a c c t c

BsgI 1320 210 215 220 225 VDAKAILSARLSKSRRLENL Cas9 210 215 220 225 VDAKAILSARLSKSRRLENL Cas9 VDAKAILSARLSKSRRLENL t a g a t g c t a a a g c g a t t c t t t c t g c a c g a t t g a g t a a a t c a a g a c g a t t a g a a a a t c t c a a t c t a c g a t t t c g c t a a g a a a g a c g t g c t a a c t c a t t t a g t t c t g c t a a t c t t t t a g a g t

1380 230 235 240 245 IAQLPGEKKNGLFGNLIALS Cas9 230 235 240 245 IAQLPGEKKNGLFGNLIALS Cas9 IAQLPGEKKNGLFGNLIALS t t g c t c a g c t c c c c g g t g a g a a g a a a a a t g g c t t a t t t g g g a a t c t c a t t g c t t t g t c a t a a c g a g t c g a g g g g c c a c t c t t c t t t t t a c c g a a t a a a c c c t t a g a g t a a c g a a a c a g t a

BstAPI 1440 250 255 260 265 LGLTPNFKSNFDLAEDAKLQ Cas9 250 255 260 265 LGLTPNFKSNFDLAEDAKLQ Cas9 LGLTPNFKSNFDLAEDAKLQ t g g g t t t g a c c c c t a a t t t t a a a t c a a a t t t t g a t t t g g c a g a a g a t g c t a a a t t a c a g c a c c c a a a c t g g g g a t t a a a a t t t a g t t t a a a a c t a a a c c g t c t t c t a c g a t t t a a t g t c g

1500 270 275 280 285 LSKDTYDDDLDNLLAQIGDQ Cas9 270 275 280 285 LSKDTYDDDLDNLLAQIGDQ Cas9 LSKDTYDDDLDNLLAQIGDQ t t t c a a a a g a t a c t t a c g a t g a t g a t t t a g a t a a t t t a t t g g c g c a a a t t g g a g a t c a a t a a a g t t t t c t a t g a a t g c t a c t a c t a a a t c t a t t a a a t a a c c g c g t t t a a c c t c t a g t t a

1560 290 295 300 305 YADLFLAAKNLSDAILLSDI Cas9 290 295 300 305 YADLFLAAKNLSDAILLSDI Cas9 YADLFLAAKNLSDAILLSDI a t g c t g a t t t g t t t t t g g c a g c t a a g a a t t t a t c a g a t g c t a t t t t a c t t t c a g a t a t c c t a c g a c t a a a c a a a a a c c g t c g a t t c t t a a a t a g t c t a c g a t a a a a t g a a a g t c t a t a g g

1620 310 315 320 325 LRVNTEITKAPLSASMIKRY Cas9 310 315 320 325 LRVNTEITKAPLSASMIKRY Cas9 LRVNTEITKAPLSASMIKRY t a a g a g t a a a t a c t g a a a t a a c t a a g g c t c c c c t a t c a g c t t c a a t g a t t a a a c g c t a c g a t t c t c a t t t a t g a c t t t a t t g a t t c c g a g g g g a t a g t c g a a g t t a c t a a t t t g c g a t g c

1680 330 335 340 345 DEHHQDLTLLKALVRQQLPE Cas9 330 335 340 345 DEHHQDLTLLKALVRQQLPE Cas9 DEHHQDLTLLKALVRQQLPE a t g a a c a t c a t c a a g a c t t g a c t c t t t t a a a a g c t t t a g t t c g a c a a c a a c t t c c a g a a a t a c t t g t a g t a g t t c t g a a c t g a g a a a a t t t t c g a a a t c a a g c t g t t g t t g a a g g t c t t t

BspMIBfuAI 1740 350 355 360 365 KYKEIFFDQSKNGYAGYIDG Cas9 350 355 360 365 KYKEIFFDQSKNGYAGYIDG Cas9 KYKEIFFDQSKNGYAGYIDG a g t a t a a a g a a a t c t t t t t t g a t c a a t c a a a a a a c g g a t a t g c a g g t t a t a t t g a t g g g g t c a t a t t t c t t t a g a a a a a a c t a g t t a g t t t t t t g c c t a t a c g t c c a a t a t a a c t a c c c c

BmtINheI 1800 370 375 380 385 GASQEEFYKFIKPILEKMDG Cas9 370 375 380 385 GASQEEFYKFIKPILEKMDG Cas9 GASQEEFYKFIKPILEKMDG g a g c t a g c c a a g a a g a a t t t t a t a a a t t t a t c a a a c c a a t t t t a g a a a a a a t g g a t g g t a c t c g a t c g g t t c t t c t t a a a a t a t t t a a a t a g t t t g g t t a a a a t c t t t t t t a c c t a c c a t

1860 390 395 400 405 TEELLVKLNREDLLRKQRTF Cas9 390 395 400 405 TEELLVKLNREDLLRKQRTF Cas9 TEELLVKLNREDLLRKQRTF c t g a g g a a t t a t t g g t g a a a c t a a a t c g t g a a g a t t t g c t g c g c a a g c a a c g g a c c t t t g g a c t c c t t a a t a a c c a c t t t g a t t t a g c a c t t c t a a a c g a c g c g t t c g t t g c c t g g a a a c

SphIAleI 1920 410 415 420 425 DNGSIPHQIHLGELHAILRR Cas9 410 415 420 425 DNGSIPHQIHLGELHAILRR Cas9 DNGSIPHQIHLGELHAILRR a c a a c g g c t c t a t t c c c c a t c a a a t t c a c t t g g g t g a g c t g c a t g c t a t t t t g a g a a g a c t g t t g c c g a g a t a a g g g g t a g t t t a a g t g a a c c c a c t c g a c g t a c g a t a a a a c t c t t c t g

1980 430 435 440 445 QEDFYPFLKDNREKIEKILT Cas9 430 435 440 445 QEDFYPFLKDNREKIEKILT Cas9 QEDFYPFLKDNREKIEKILT a a g a a g a c t t t t a t c c a t t t t t a a a a g a c a a t c g t g a g a a g a t t g a a a a a a t c t t g a c t t t t c t t c t g a a a a t a g g t a a a a a t t t t c t g t t a g c a c t c t t c t a a c t t t t t t a g a a c t g a a

2040 450 455 460 465 FRIPYYVGPLARGNSRFAWM Cas9 450 455 460 465 FRIPYYVGPLARGNSRFAWM Cas9 FRIPYYVGPLARGNSRFAWM t t c g a a t t c c t t a t t a t g t t g g t c c a t t g g c g c g t g g c a a t a g t c g t t t t g c a t g g a t g a a a g c t t a a g g a a t a a t a c a a c c a g g t a a c c g c g c a c c g t t a t c a g c a a a a c g t a c c t a c t

2100 470 475 480 485 TRKSEETITPWNFEEVVDKG Cas9 470 475 480 485 TRKSEETITPWNFEEVVDKG Cas9 TRKSEETITPWNFEEVVDKG c t c g g a a g t c t g a a g a a a c a a t t a c c c c a t g g a a t t t t g a a g a a g t t g t c g a t a a a g g t g g a g c c t t c a g a c t t c t t t g t t a a t g g g g t a c c t t a a a a c t t c t t c a a c a g c t a t t t c c a c

2160 490 495 500 505 ASAQSFIERMTNFDKNLPNE Cas9 490 495 500 505 ASAQSFIERMTNFDKNLPNE Cas9 ASAQSFIERMTNFDKNLPNE c t t c a g c t c a a t c a t t t a t t g a a c g c a t g a c a a a c t t t g a t a a a a a t c t t c c a a a t g a a a g a a g t c g a g t t a g t a a a t a a c t t g c g t a c t g t t t g a a a c t a t t t t t a g a a g g t t t a c t t t

2220 510 515 520 525 KVLPKHSLLYEYFTVYNELT Cas9 510 515 520 525 KVLPKHSLLYEYFTVYNELT Cas9 KVLPKHSLLYEYFTVYNELT a a g t a c t a c c a a a a c a t a g t t t g c t t t a t g a g t a t t t t a c g g t t t a t a a c g a a t t g a c a a t t c a t g a t g g t t t t g t a t c a a a c g a a a t a c t c a t a a a a t g c c a a a t a t t g c t t a a c t g t t

BsmI 2280 530 535 540 545 KVKYVTEGMRKPAFLSGEQK Cas9 530 535 540 545 KVKYVTEGMRKPAFLSGEQK Cas9 KVKYVTEGMRKPAFLSGEQK a g g t c a a a t a t g t t a c t g a a g g a a t g c g a a a a c c a g c a t t t c t t t c a g g t g a a c a g a a g a t c c a g t t t a t a c a a t g a c t t c c t t a c g c t t t t g g t c g t a a a g a a a g t c c a c t t g t c t t c t

2340 550 555 560 565 KAIVDLLFKTNRKVTVKQLK Cas9 550 555 560 565 KAIVDLLFKTNRKVTVKQLK Cas9 KAIVDLLFKTNRKVTVKQLK a a g c c a t t g t t g a t t t a c t c t t c a a a a c a a a t c g a a a a g t a a c c g t t a a g c a a t t a a a a g t t c g g t a a c a a c t a a a t g a g a a g t t t t g t t t a g c t t t t c a t t g g c a a t t c g t t a a t t t t c

2400 570 575 580 585 EDYFKKIECFDSVEISGVED Cas9 570 575 580 585 EDYFKKIECFDSVEISGVED Cas9 EDYFKKIECFDSVEISGVED a a g a t t a t t t c a a a a a a a t a g a a t g t t t t g a t a g t g t t g a a a t t t c a g g a g t t g a a g a t a t t c t a a t a a a g t t t t t t t a t c t t a c a a a a c t a t c a c a a c t t t a a a g t c c t c a a c t t c t a t

Acc65IKpnI 2460 590 595 600 605 RFNASLGTYHDLLKIIKDKD Cas9 590 595 600 605 RFNASLGTYHDLLKIIKDKD Cas9 RFNASLGTYHDLLKIIKDKD g a t t t a a t g c t t c a t t a g g t a c c t a c c a t g a t t t g c t a a a a a t t a t t a a a g a t a a a g a t t c t a a a t t a c g a a g t a a t c c a t g g a t g g t a c t a a a c g a t t t t t a a t a a t t t c t a t t t c t a a

2520 610 615 620 625 FLDNEENEDILEDIVLTLTL Cas9 610 615 620 625 FLDNEENEDILEDIVLTLTL Cas9 FLDNEENEDILEDIVLTLTL t t t t g g a t a a t g a a g a a a a t g a a g a t a t c t t a g a g g a t a t t g t t t t a a c a t t g a c c t t a t a a a a c c t a t t a c t t c t t t t a c t t c t a t a g a a t c t c c t a t a a c a a a a t t g t a a c t g g a a t a

2580 630 635 640 645 FEDREMIEERLKTYAHLFDD Cas9 630 635 640 645 FEDREMIEERLKTYAHLFDD Cas9 FEDREMIEERLKTYAHLFDD t t g a a g a t a g g g a g a t g a t t g a g g a a a g a c t t a a a a c a t a t g c t c a c c t c t t t g a t g a t a a a c t t c t a t c c c t c t a c t a a c t c c t t t c t g a a t t t t g t a t a c g a g t g g a g a a a c t a c t a t

2640 650 655 660 665 KVMKQLKRRRYTGWGRLSRK Cas9 650 655 660 665 KVMKQLKRRRYTGWGRLSRK Cas9 KVMKQLKRRRYTGWGRLSRK a g g t g a t g a a a c a g c t t a a a c g t c g c c g t t a t a c t g g t t g g g g a c g t t t g t c t c g a a a a t t c c a c t a c t t t g t c g a a t t t g c a g c g g c a a t a t g a c c a a c c c c t g c a a a c a g a g c t t t t a

2700 670 675 680 685 LINGIRDKQSGKTILDFLKS Cas9 670 675 680 685 LINGIRDKQSGKTILDFLKS Cas9 LINGIRDKQSGKTILDFLKS t g a t t a a t g g t a t t a g g g a t a a g c a a t c t g g c a a a a c a a t a t t a g a t t t t t t g a a a t c a g a c t a a t t a c c a t a a t c c c t a t t c g t t a g a c c g t t t t g t t a t a a t c t a a a a a a c t t t a g t c

2760 690 695 700 705 DGFANRNFMQLIHDDSLTFK Cas9 690 695 700 705 DGFANRNFMQLIHDDSLTFK Cas9 DGFANRNFMQLIHDDSLTFK a t g g t t t t g c c a a t c g c a a t t t t a t g c a g c t g a t c c a t g a t g a t a g t t t g a c a t t t a a a g t a c c a a a a c g g t t a g c g t t a a a a t a c g t c g a c t a g g t a c t a c t a t c a a a c t g t a a a t t t c

2820 710 715 720 725 EDIQKAQVSGQGDSLHEHIA Cas9 710 715 720 725 EDIQKAQVSGQGDSLHEHIA Cas9 EDIQKAQVSGQGDSLHEHIA a a g a c a t t c a a a a a g c a c a a g t g t c t g g a c a a g g c g a t a g t t t a c a t g a a c a t a t t g c a a t t c t g t a a g t t t t t c g t g t t c a c a g a c c t g t t c c g c t a t c a a a t g t a c t t g t a t a a c g t t

2880 730 735 740 745 NLAGSPAIKKGILQTVKVVD Cas9 730 735 740 745 NLAGSPAIKKGILQTVKVVD Cas9 NLAGSPAIKKGILQTVKVVD a t t t a g c t g g t a g c c c t g c t a t t a a a a a a g g t a t t t t a c a g a c t g t a a a a g t t g t t g a t g t a a a t c g a c c a t c g g g a c g a t a a t t t t t t c c a t a a a a t g t c t g a c a t t t t c a a c a a c t a c

2940 750 755 760 765 ELVKVMGRHKPENIVIEMAR Cas9 750 755 760 765 ELVKVMGRHKPENIVIEMAR Cas9 ELVKVMGRHKPENIVIEMAR a a t t g g t c a a a g t a a t g g g g c g g c a t a a g c c a g a a a a t a t c g t t a t t g a a a t g g c a c g t g t t a a c c a g t t t c a t t a c c c c g c c g t a t t c g g t c t t t t a t a g c a a t a a c t t t a c c g t g c a c

3000 770 775 780 785 ENQTTQKGQKNSRERMKRIE Cas9 770 775 780 785 ENQTTQKGQKNSRERMKRIE Cas9 ENQTTQKGQKNSRERMKRIE a a a a t c a g a c a a c t c a a a a g g g c c a g a a a a a t t c g c g a g a g c g t a t g a a a c g a a t c g a a g t t t t a g t c t g t t g a g t t t t c c c g g t c t t t t t a a g c g c t c t c g c a t a c t t t g c t t a g c t t c

3060 790 795 800 805 EGIKELGSQILKEHPVENTQ Cas9 790 795 800 805 EGIKELGSQILKEHPVENTQ Cas9 EGIKELGSQILKEHPVENTQ a a g g t a t c a a a g a a t t a g g a a g t c a g a t t c t t a a a g a g c a t c c t g t t g a a a a t a c t c a a t t t c c a t a g t t t c t t a a t c c t t c a g t c t a a g a a t t t c t c g t a g g a c a a c t t t t a t g a g t t a

3120 810 815 820 825 LQNEKLYLYYLQNGRDMYVD Cas9 810 815 820 825 LQNEKLYLYYLQNGRDMYVD Cas9 LQNEKLYLYYLQNGRDMYVD t g c a a a a t g a a a a g c t c t a t c t c t a t t a t c t c c a a a a t g g a a g a g a c a t g t a t g t g g a c c a c g t t t t a c t t t t c g a g a t a g a g a t a a t a g a g g t t t t a c c t t c t c t g t a c a t a c a c c t g g

3180 830 835 840 845 QELDINRLSDYDVDHIVPQS Cas9 830 835 840 845 QELDINRLSDYDVDHIVPQS Cas9 QELDINRLSDYDVDHIVPQS a a g a a t t a g a t a t t a a t c g t t t a a g t g a t t a t g a t g t c g a t c a c a t t g t t c c a c a a a g t t t t c t t a a t c t a t a a t t a g c a a a t t c a c t a a t a c t a c a g c t a g t g t a a c a a g g t g t t t c a a

3240 850 855 860 865 FLKDDSIDNKVLTRSDKNRG Cas9 850 855 860 865 FLKDDSIDNKVLTRSDKNRG Cas9 FLKDDSIDNKVLTRSDKNRG t c c t t a a a g a c g a t t c a a t a g a c a a t a a g g t c t t a a c g c g t t c t g a t a a a a a t c g t g g t a a g g a a t t t c t g c t a a g t t a t c t g t t a t t c c a g a a t t g c g c a a g a c t a t t t t t a g c a c c a t

3300 870 875 880 885 KSDNVPSEEVVKKMKNYWRQ Cas9 870 875 880 885 KSDNVPSEEVVKKMKNYWRQ Cas9 KSDNVPSEEVVKKMKNYWRQ a a t c g g a t a a c g t t c c a a g t g a a g a a g t a g t c a a a a a g a t g a a a a a c t a t t g g a g a c a a c t t a g c c t a t t g c a a g g t t c a c t t c t t c a t c a g t t t t t c t a c t t t t t g a t a a c c t c t g t t g

3360 890 895 900 905 LLNAKLITQRKFDNLTKAER Cas9 890 895 900 905 LLNAKLITQRKFDNLTKAER Cas9 LLNAKLITQRKFDNLTKAER t t c t a a a c g c c a a g t t a a t c a c t c a a c g t a a g t t t g a t a a t t t a a c g a a a g c t g a a c g t g a a g a t t t g c g g t t c a a t t a g t g a g t t g c a t t c a a a c t a t t a a a t t g c t t t c g a c t t g c a c

3420 910 915 920 925 GGLSELDKAGFIKRQLVETR Cas9 910 915 920 925 GGLSELDKAGFIKRQLVETR Cas9 GGLSELDKAGFIKRQLVETR g a g g t t t g a g t g a a c t t g a t a a a g c t g g t t t t a t c a a a c g c c a a t t g g t t g a a a c t c g c c c t c c a a a c t c a c t t g a a c t a t t t c g a c c a a a a t a g t t t g c g g t t a a c c a a c t t t g a g c g g

3480 930 935 940 945 QITKHVAQILDSRMNTKYDE Cas9 930 935 940 945 QITKHVAQILDSRMNTKYDE Cas9 QITKHVAQILDSRMNTKYDE a a a t c a c t a a g c a t g t g g c a c a a a t t t t g g a t a g t c g c a t g a a t a c t a a a t a c g a t g a a a t t t a g t g a t t c g t a c a c c g t g t t t a a a a c c t a t c a g c g t a c t t a t g a t t t a t g c t a c t t t

3540 950 955 960 965 NDKLIREVKVITLKSKLVSD Cas9 950 955 960 965 NDKLIREVKVITLKSKLVSD Cas9 NDKLIREVKVITLKSKLVSD a t g a t a a a c t t a t t c g a g a g g t t a a a g t g a t t a c c t t a a a a t c t a a a t t a g t t t c t g a c t t a c t a t t t g a a t a a g c t c t c c a a t t t c a c t a a t g g a a t t t t a g a t t t a a t c a a a g a c t g a

3600 970 975 980 985 FRKDFQFYKVREINNYHHAH Cas9 970 975 980 985 FRKDFQFYKVREINNYHHAH Cas9 FRKDFQFYKVREINNYHHAH t c c g a a a a g a t t t c c a a t t c t a t a a a g t a c g t g a g a t t a a c a a t t a c c a t c a t g c c c a t g a g g c t t t t c t a a a g g t t a a g a t a t t t c a t g c a c t c t a a t t g t t a a t g g t a g t a c g g g t a c

3660 990 995 1000 1005 DAYLNAVVGTALIKKYPKLE Cas9 990 995 1000 1005 DAYLNAVVGTALIKKYPKLE Cas9 DAYLNAVVGTALIKKYPKLE a t g c g t a t c t a a a t g c c g t c g t t g g a a c t g c t t t g a t t a a g a a a t a t c c a a a a c t t g a a t t a c g c a t a g a t t t a c g g c a g c a a c c t t g a c g a a a c t a a t t c t t t a t a g g t t t t g a a c t t a

3720 1010 1015 1020 1025 SEFVYGDYKVYDVRKMIAKS Cas9 1010 1015 1020 1025 SEFVYGDYKVYDVRKMIAKS Cas9 SEFVYGDYKVYDVRKMIAKS c g g a g t t t g t c t a t g g t g a t t a t a a a g t t t a t g a t g t t c g t a a a a t g a t t g c t a a g t c t g g c c t c a a a c a g a t a c c a c t a a t a t t t c a a a t a c t a c a a g c a t t t t a c t a a c g a t t c a g a c

3780 1030 1035 1040 1045 EQEIGKATAKYFFYSNIMNF Cas9 1030 1035 1040 1045 EQEIGKATAKYFFYSNIMNF Cas9 EQEIGKATAKYFFYSNIMNF a g c a a g a a a t a g g c a a a g c a a c c g c a a a a t a t t t c t t t t a c t c t a a t a t c a t g a a c t t c t t c g t t c t t t a t c c g t t t c g t t g g c g t t t t a t a a a g a a a a t g a g a t t a t a g t a c t t g a a g a

3840 1050 1055 1060 1065 FKTEITLANGEIRKRPLIET Cas9 1050 1055 1060 1065 FKTEITLANGEIRKRPLIET Cas9 FKTEITLANGEIRKRPLIET t c a a a a c a g a a a t t a c a c t t g c a a a t g g a g a g a t t c g c a a a c g c c c t c t a a t c g a a a c t a a g t t t t g t c t t t a a t g t g a a c g t t t a c c t c t c t a a g c g t t t g c g g g a g a t t a g c t t t g a t

3900 1070 1075 1080 1085 NGETGEIVWDKGRDFATVRK Cas9 1070 1075 1080 1085 NGETGEIVWDKGRDFATVRK Cas9 NGETGEIVWDKGRDFATVRK a t g g g g a a a c t g g a g a a a t t g t c t g g g a t a a a g g g c g a g a t t t t g c c a c a g t g c g c a a a g t a c c c c t t t g a c c t c t t t a a c a g a c c c t a t t t c c c g c t c t a a a a c g g t g t c a c g c g t t t c

3960 1090 1095 1100 1105 VLSMPQVNIVKKTEVQTGGF Cas9 1090 1095 1100 1105 VLSMPQVNIVKKTEVQTGGF Cas9 VLSMPQVNIVKKTEVQTGGF t a t t g t c c a t g c c c c a a g t c a a t a t t g t c a a g a a a a c a g a a g t a c a g a c a g g c g g a t t c t a t a a c a g g t a c g g g g t t c a g t t a t a a c a g t t c t t t t g t c t t c a t g t c t g t c c g c c t a a g a

4020 1110 1115 1120 1125 SKESILPKRNSDKLIARKKD Cas9 1110 1115 1120 1125 SKESILPKRNSDKLIARKKD Cas9 SKESILPKRNSDKLIARKKD c c a a g g a g t c a a t t t t a c c a a a a a g a a a t t c g g a c a a g c t t a t t g c t c g t a a a a a a g a c t g g t t c c t c a g t t a a a a t g g t t t t t c t t t a a g c c t g t t c g a a t a a c g a g c a t t t t t t c t g a

4080 1130 1135 1140 1145 WDPKKYGGFDSPTVAYSVLV Cas9 1130 1135 1140 1145 WDPKKYGGFDSPTVAYSVLV Cas9 WDPKKYGGFDSPTVAYSVLV g g g a t c c a a a a a a a t a t g g t g g t t t t g a t a g t c c a a c g g t a g c t t a t t c a g t c c t a g t g g c c c t a g g t t t t t t t a t a c c a c c a a a a c t a t c a g g t t g c c a t c g a a t a a g t c a g g a t c a c c

4140 1150 1155 1160 1165 VAKVEKGKSKKLKSVKELLG Cas9 1150 1155 1160 1165 VAKVEKGKSKKLKSVKELLG Cas9 VAKVEKGKSKKLKSVKELLG t t g c t a a g g t g g a a a a a g g g a a a t c g a a g a a g t t a a a a t c c g t t a a a g a g t t a c t a g g g a a a c g a t t c c a c c t t t t t c c c t t t a g c t t c t t c a a t t t t a g g c a a t t t c t c a a t g a t c c c t

4200 1170 1175 1180 1185 ITIMERSSFEKNPIDFLEAK Cas9 1170 1175 1180 1185 ITIMERSSFEKNPIDFLEAK Cas9 ITIMERSSFEKNPIDFLEAK t c a c a a t t a t g g a a a g a a g t t c c t t t g a a a a a a a t c c g a t t g a c t t t t t a g a a g c t a a a g a g t g t t a a t a c c t t t c t t c a a g g a a a c t t t t t t t a g g c t a a c t g a a a a a t c t t c g a t t t c

4260 1190 1195 1200 1205 GYKEVKKDLIIKLPKYSLFE Cas9 1190 1195 1200 1205 GYKEVKKDLIIKLPKYSLFE Cas9 GYKEVKKDLIIKLPKYSLFE g a t a t a a g g a a g t t a a a a a a g a c t t a a t c a t t a a a c t a c c t a a a t a t a g t c t t t t t g a g t c t a t a t t c c t t c a a t t t t t t c t g a a t t a g t a a t t t g a t g g a t t t a t a t c a g a a a a a c t c a

4320 1210 1215 1220 1225 LENGRKRMLASAGELQKGNE Cas9 1210 1215 1220 1225 LENGRKRMLASAGELQKGNE Cas9 LENGRKRMLASAGELQKGNE t a g a a a a c g g t c g t a a a c g g a t g c t g g c t a g t g c c g g a g a a t t a c a a a a a g g a a a t g a g c a t c t t t t g c c a g c a t t t g c c t a c g a c c g a t c a c g g c c t c t t a a t g t t t t t c c t t t a c t c g

4380 1230 1235 1240 1245 LALPSKYVNFLYLASHYEKL Cas9 1230 1235 1240 1245 LALPSKYVNFLYLASHYEKL Cas9 LALPSKYVNFLYLASHYEKL t g g c t c t g c c a a g c a a a t a t g t g a a t t t t t t a t a t t t a g c t a g t c a t t a t g a a a a g t t g a a c c g a g a c g g t t c g t t t a t a c a c t t a a a a a a t a t a a a t c g a t c a g t a a t a c t t t t c a a c t

4440 1250 1255 1260 1265 KGSPEDNEQKQLFVEQHKHY Cas9 1250 1255 1260 1265 KGSPEDNEQKQLFVEQHKHY Cas9 KGSPEDNEQKQLFVEQHKHY a g g g t a g t c c a g a a g a t a a c g a a c a a a a a c a a t t g t t t g t g g a g c a g c a t a a g c a t t a t t t c c c a t c a g g t c t t c t a t t g c t t g t t t t t g t t a a c a a a c a c c t c g t c g t a t t c g t a a t a a

4500 1270 1275 1280 1285 LDEIIEQISEFSKRVILADA Cas9 1270 1275 1280 1285 LDEIIEQISEFSKRVILADA Cas9 LDEIIEQISEFSKRVILADA t a g a t g a g a t t a t t g a g c a a a t c a g t g a a t t t t c t a a g c g t g t t a t t t t a g c a g a t g c c a a t c t a c t c t a a t a a c t c g t t t a g t c a c t t a a a a g a t t c g c a c a a t a a a a t c g t c t a c g g t

4560 1290 1295 1300 1305 NLDKVLSAYNKHRDKPIREQ Cas9 1290 1295 1300 1305 NLDKVLSAYNKHRDKPIREQ Cas9 NLDKVLSAYNKHRDKPIREQ a t t t a g a t a a a g t t c t t a g t g c a t a t a a c a a a c a t a g a g a c a a a c c a a t a c g t g a a c a a g t a a a t c t a t t t c a a g a a t c a c g t a t a t t g t t t g t a t c t c t g t t t g g t t a t g c a c t t g t t c

4620 1310 1315 1320 1325 AENIIHLFTLTNLGAPAAFK Cas9 1310 1315 1320 1325 AENIIHLFTLTNLGAPAAFK Cas9 AENIIHLFTLTNLGAPAAFK c a g a a a a t a t t a t t c a t t t a t t t a c g t t g a c g a a t c t t g g a g c t c c c g c t g c t t t t a a a t g t c t t t t a t a a t a a g t a a a t a a a t g c a a c t g c t t a g a a c c t c g a g g g c g a c g a a a a t t t a

4680 1330 1335 1340 1345 YFDTTIDRKRYTSTKEVLDA Cas9 1330 1335 1340 1345 YFDTTIDRKRYTSTKEVLDA Cas9 YFDTTIDRKRYTSTKEVLDA a t t t t g a t a c a a c a a t t g a t c g t a a a c g a t a t a c g t c t a c a a a a g a a g t t t t a g a t g c c a t a a a a c t a t g t t g t t a a c t a g c a t t t g c t a t a t g c a g a t g t t t t c t t c a a a a t c t a c g g t

4740 1350 1355 1360 1365 TLIHQSITGLYETRIDLSQL Cas9 1350 1355 1360 1365 TLIHQSITGLYETRIDLSQL Cas9 TLIHQSITGLYETRIDLSQL c t c t t a t c c a t c a a t c c a t c a c t g g t c t t t a t g a a a c a c g c a t t g a t t t g a g t c a g c t a g g a g a a t a g g t a g t t a g g t a g t g a c c a g a a a t a c t t t g t g c g t a a c t a a a c t c a g t c g a t c

4800 GGD GS 130 PKKKRKV (in frame with SV40 NLS) SSGGAAG* Cas9 SV40 NLS GGD (in frame with Cas9) GSPKKKRKVSSGGAAG* Cas9 GGDGSPKKKRKVSSGGAAG* g a g g t g a c g g t t c t c c c a a g a a g a a g a g g a a a g t c t c g a g c g g t g g a g c t g c a g g a t g a g c t c c a c t g c c a a g a g g g t t c t t c t t c t c c t t t c a g a g c t c g c c a c c t c g a c g t c c t a c t c

BsrGISacIIBsiWINotI 4860 c g g c c g c c t g c a g c t g g c g c c a t c g a t a c g c g t a c g t c g c g a c c g c g g a c a t g t a c a g a g g c c g g c g g a c g t c g a c c g c g g t a g c t a t g c g c a t g c a g c g c t g g c g c c t g t a c a t g t c t c

4920 c t c g a g a t g a t c g g t t t g t g t t c c t a c t t a g t t a a t t g t g a t a g t t t a t t t t t g t a a a t a g a g c t c t a c t a g c c a a a c a c a a g g a t g a a t c a a t t a a c a c t a t c a a a t a a a a a c a t t t a t

SnaBI 4980 a t a g t a c g t a a a g t t a c t t t a g t t a t a a t a a a g t a g a t a g t g a c t g c a t t a a g t t t a a a g t a t c a t g c a t t t c a a t g a a a t c a a t a t t a t t t c a t c t a t c a c t g a c g t a a t t c a a a t t t c

5040 a c t g a t t t t t g t a g a t t a t t t a a a a a t t t t t a a t a a a t t a t t t a a t t t a a a g a t a a a a a g t g a c t a a a a a c a t c t a a t a a a t t t t t a a a a a t t a t t t a a t a a a t t a a a t t t c t a t t t t t c

5100 t t g t t t c a t t t t a a a a t a a t a a a a a a c t t a c c g c a t t a g t c g t g g c c t t g g c a g t a t t a a a a c a a a g t a a a a t t t t a t t a t t t t t t g a a t g g c g t a a t c a g c a c c g g a a c c g t c a t a a t t

5160 g c g c a a a c g g g c t c t c c t t c g t a t g c c c a t c a g g a a t a t a g a a a a a a a c a a c t g g c t g c t c g c g t t t g c c c g a g a g g a a g c a t a c g g g t a g t c c t t a t a t c t t t t t t t g t t g a c c g a c g a

XcmIAflIISfiI 5220 c a c t a g t g g c c a c g t g g g c c g t g c a c c t t a a g c t t g g c g c g c c t a g g c c g g c c a g c t t g g g t g a t c a c c g g t g c a c c c g g c a c g t g g a a t t c g a a c c g c g c g g a t c c g g c c g g t c g a a c c

5280 M13 fwd T G A C C G G C A G C A A A A T G T M13 Forward c a c t g g c c g t c g t t t t a c a a c g t c g t g a c t g g g a a a a c c c t g g c g t t a c c c a a c t t a a t c g t g a c c g g c a g c a a a a t g t t g c a g c a c t g a c c c t t t t g g g a c c g c a a t g g g t t g a a t t a g

5340 g c c t t g c a g c a c a t c c c c c t t t c g c c a g c t g g c g t a a t a g c g a a g a g g c c c g c a c c g a t c c g g a a c g t c g t g t a g g g g g a a a g c g g t c g a c c g c a t t a t c g c t t c t c c g g g c g t g g c t a g

5400 g c c c t t c c c a a c a g t t g c g c a g c c t g a a t g g c g a a t g g c g c c t g a t g c g g t a t t t t c t c c c g g g a a g g g t t g t c a a c g c g t c g g a c t t a c c g c t t a c c g c g g a c t a c g c c a t a a a a g a g g

5460 f1 ori t t a c g c a t c t g t g c g g t a t t t c a c a c c g c a t a c g t c a a a g c a a c c a t a g t a c g c g c c c t g a a t g c g t a g a c a c g c c a t a a a g t g t g g c g t a t g c a g t t t c g t t g g t a t c a t g c g c g g g a c

5520 f1 ori t a g c g g c g c a t t a a g c g c g g c g g g t g t g g t g g t t a c g c g c a g c g t g a c c g c t a c a c t t g c a t c g c c g c g t a a t t c g c g c c g c c c a c a c c a c c a a t g c g c g t c g c a c t g g c g a t g t g a a c g

5580 f1 ori G A G G A A A G C G A A A G A A G G G A F1ori-R c a g c g c c c t a g c g c c c g c t c c t t t c g c t t t c t t c c c t t c c t t t c t c g c c a c g t t c g c c g g g t c g c g g g a t c g c g g g c g a g g a a a g c g a a a g a a g g g a a g g a a a g a g c g g t g c a a g c g g c c

5640 f1 ori c t t t c c c c g t c a a g c t c t a a a t c g g g g g c t c c c t t t a g g g t t c c g a t t t a g t g c t t t a c g g a a a g g g g c a g t t c g a g a t t t a g c c c c c g a g g g a a a t c c c a a g g c t a a a t c a c g a a a t g c

DraIIIBtgZI 5700 f1 ori g c a c c t c g a c c c c a a a a a a c t t g a t t t g g g t g a t g g t t c a c g t a g t g g g c c a t c g c c c t g c g t g g a g c t g g g g t t t t t t g a a c t a a a c c c a c t a c c a a g t g c a t c a c c c g g t a g c g g g a c

5760 f1 ori G T G G A C T C T T G T T F1ori-F a t a g a c g g t t t t t c g c c c t t t g a c g t t g g a g t c c a c g t t c t t t a a t a g t g g a c t c t t g t t t a t c t g c c a a a a a g c g g g a a a c t g c a a c c t c a g g t g c a a g a a a t t a t c a c c t g a g a a c a a

5820 f1 ori C C A A A C T G G F1ori-F c c a a a c t g g a a c a a c a c t c a a c c c t a t c t c g g g c t a t t c t t t t g a t t t a t a a g g g a t t t t g g t t t g a c c t t g t t g t g a g t t g g g a t a g a g c c c g a t a a g a a a a c t a a a t a t t c c c t a a a a

5880 f1 ori g c c g a t t t c g g c c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a t t t a a c g c g a a t t t c g g c t a a a g c c g g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t a a a t t g c g c t t a a a

5940 f1 ori A T G T T A G A C G A G A C T A C G pRS-marker t a a c a a a a t a t t a a c g t t t a c a a t t t t a t g g t g c a c t c t c a g t a c a a t c t g c t c t g a t g c a t t g t t t t a t a a t t g c a a a t g t t a a a a t a c c a c g t g a g a g t c a t g t t a g a c g a g a c t a c g

6000 G C pRS-marker c g c a t a g t t a a g c c a g c c c c g a c a c c c g c c a a c a c c c g c t g a c g c g c c c t g a c g g g c t t g g c g t a t c a a t t c g g t c g g g g c t g t g g g c g g t t g t g g g c g a c t g c g c g g g a c t g c c c g a a c

PfoI 6060 C C G G G A G C T G C A T G T G T C A pGEX 3' t c t g c t c c c g g c a t c c g c t t a c a g a c a a g c t g t g a c c g t c t c c g g g a g c t g c a t g t g t c a a g a c g a g g g c c g t a g g c g a a t g t c t g t t c g a c a c t g g c a g a g g c c c t c g a c g t a c a c a g t

6120 G A G G pGEX 3' C G G A G C A C T A T G C G G A T A A pBRforEco g a g g t t t t c a c c g t c a t c a c c g a a a c g c g c g a g a c g a a a g g g c c t c g t g a t a c g c c t a t t c t c c a a a a g t g g c a g t a g t g g c t t t g c g c g c t c t g c t t t c c c g g a g c a c t a t g c g g a t a a

6180 t t t a t a g g t t a a t g t c a t g a t a a t a a t g g t t t c t t a g a c g t c a g g t g g c a c t t t t c g g g g a a a t a t c c a a t t a c a g t a c t a t t a t t a c c a a a g a a t c t g c a g t c c a c c g t g a a a a g c c c c

6240 AmpR promoter a a a t g t g c g c g g a a c c c c t a t t t g t t t a t t t t t c t a a a t a c a t t c a a a t a t g t a t c c g c t t t t a c a c g c g c c t t g g g g a t a a a c a a a t a a a a a g a t t t a t g t a a g t t t a t a c a t a g g c g a

6300 1 signal sequence MSI AmpR promoter AmpR MSI c a t g a g a c a a t a a c c c t g a t a a a t g c t t c a a t a a t a t t g a a a a a g g a a g a g t a t g a g t a t g t a c t c t g t t a t t g g g a c t a t t t a c g a a g t t a t t a t a a c t t t t t c c t t c t c a t a c t c a t a

6360 5 10 15 20 signal sequence QHFRVALIPFFAAFCLPVFA AmpR QHFRVALIPFFAAFCLPVFA t c a a c a t t t c c g t g t c g c c c t t a t t c c c t t t t t t g c g g c a t t t t g c c t t c c t g t t t t t g c a g t t g t a a a g g c a c a g c g g g a a t a a g g g a a a a a a c g c c g t a a a a c g g a a g g a c a a a a a c g

6420 25 30 35 40 HPETLVKVKDAEDQLGARVG AmpR HPETLVKVKDAEDQLGARVG t c a c c c a g a a a c g c t g g t g a a a g t a a a a g a t g c t g a a g a t c a g t t g g g t g c a c g a g t g g g a g t g g g t c t t t g c g a c c a c t t t c a t t t t c t a c g a c t t c t a g t c a a c c c a c g t g c t c a c c c

6480 45 50 55 60 YIELDLNSGKILESFRPEER AmpR YIELDLNSGKILESFRPEER t t a c a t c g a a c t g g a t c t c a a c a g c g g t a a g a t c c t t g a g a g t t t t c g c c c c g a a g a a c g a a t g t a g c t t g a c c t a g a g t t g t c g c c a t t c t a g g a a c t c t c a a a a g c g g g g c t t c t t g c

6540 65 70 75 80 FPMMSTFKVLLCGAVLSRID AmpR C G A T A C A C C G C G C C A T A A T A Amp-R FPMMSTFKVLLCGAVLSRID t t t t c c a a t g a t g a g c a c t t t t a a a g t t c t g c t a t g t g g c g c g g t a t t a t c c c g t a t t g a a a a a g g t t a c t a c t c g t g a a a a t t t c a a g a c g a t a c a c c g c g c c a t a a t a g g g c a t a a c t

6600 85 90 95 100 AGQEQLGRRIHYSQNDLVEY AmpR AGQEQLGRRIHYSQNDLVEY c g c c g g g c a a g a g c a a c t c g g t c g c c g c a t a c a c t a t t c t c a g a a t g a c t t g g t t g a g t a g c g g c c c g t t c t c g t t g a g c c a g c g g c g t a t g t g a t a a g a g t c t t a c t g a a c c a a c t c a t

6660 105 110 115 120 SPVTEKHLTDGMTVRELCSA AmpR SPVTEKHLTDGMTVRELCSA c t c a c c a g t c a c a g a a a a g c a t c t t a c g g a t g g c a t g a c a g t a a g a g a a t t a t g c a g t g c g a g t g g t c a g t g t c t t t t c g t a g a a t g c c t a c c g t a c t g t c a t t c t c t t a a t a c g t c a c g

6720 125 130 135 140 AITMSDNTAANLLLTTIGGP AmpR AITMSDNTAANLLLTTIGGP t g c c a t a a c c a t g a g t g a t a a c a c t g c g g c c a a c t t a c t t c t g a c a a c g a t c g g a g g a c c a c g g t a t t g g t a c t c a c t a t t g t g a c g c c g g t t g a a t g a a g a c t g t t g c t a g c c t c c t g g

6780 145 150 155 160 KELTAFLHNMGDHVTRLDRW AmpR KELTAFLHNMGDHVTRLDRW g a a g g a g c t a a c c g c t t t t t t g c a c a a c a t g g g g g a t c a t g t a a c t c g c c t t g a t c g t t g c t t c c t c g a t t g g c g a a a a a a c g t g t t g t a c c c c c t a g t a c a t t g a g c g g a a c t a g c a a c

6840 165 170 175 180 EPELNEAIPNDERDTTMPVA AmpR EPELNEAIPNDERDTTMPVA g g a a c c g g a g c t g a a t g a a g c c a t a c c a a a c g a c g a g c g t g a c a c c a c g a t g c c t g t a g c c c t t g g c c t c g a c t t a c t t c g g t a t g g t t t g c t g c t c g c a c t g t g g t g c t a c g g a c a t c g

6900 185 190 195 200 MATTLRKLLTGELLTLASRQ AmpR MATTLRKLLTGELLTLASRQ a a t g g c a a c a a c g t t g c g c a a a c t a t t a a c t g g c g a a c t a c t t a c t c t a g c t t c c c g g c a t t a c c g t t g t t g c a a c g c g t t t g a t a a t t g a c c g c t t g a t g a a t g a g a t c g a a g g g c c g t

NmeAIII 6960 205 210 215 220 QLIDWMEADKVAGPLLRSAL AmpR QLIDWMEADKVAGPLLRSAL a c a a t t a a t a g a c t g g a t g g a g g c g g a t a a a g t t g c a g g a c c a c t t c t g c g c t c g g c c c t t g t t a a t t a t c t g a c c t a c c t c c g c c t a t t t c a a c g t c c t g g t g a a g a c g c g a g c c g g g a

7020 225 230 235 240 PAGWFIADKSGAGERGSRGI AmpR PAGWFIADKSGAGERGSRGI t c c g g c t g g c t g g t t t a t t g c t g a t a a a t c t g g a g c c g g t g a g c g t g g g t c t c g c g g t a t a g g c c g a c c g a c c a a a t a a c g a c t a t t t a g a c c t c g g c c a c t c g c a c c c a g a g c g c c a t a

AhdI 7080 245 250 255 260 IAALGPDGKPSRIVVIYTTG AmpR IAALGPDGKPSRIVVIYTTG c a t t g c a g c a c t g g g g c c a g a t g g t a a g c c c t c c c g t a t c g t a g t t a t c t a c a c g a c g g g g t a a c g t c g t g a c c c c g g t c t a c c a t t c g g g a g g g c a t a g c a t c a a t a g a t g t g c t g c c c

7140 265 270 275 280 SQATMDERNRQIAEIGASLI AmpR SQATMDERNRQIAEIGASLI g a g t c a g g c a a c t a t g g a t g a a c g a a a t a g a c a g a t c g c t g a g a t a g g t g c c t c a c t g a t c t c a g t c c g t t g a t a c c t a c t t g c t t t a t c t g t c t a g c g a c t c t a t c c a c g g a g t g a c t a

7200 285 KHW* AmpR KHW* t a a g c a t t g g t a a c t g t c a g a c c a a g t t t a c t c a t a t a t a c t t t a g a t t g a t t t a a a a c t a t t c g t a a c c a t t g a c a g t c t g g t t c a a a t g a g t a t a t a t g a a a t c t a a c t a a a t t t t g a

7260 t c a t t t t t a a t t t a a a a g g a t c t a g g t g a a g a t c c t t t t t g a t a a t c t c a t g a c c a a a a t a g t a a a a a t t a a a t t t t c c t a g a t c c a c t t c t a g g a a a a a c t a t t a g a g t a c t g g t t t t a

7320 c c c t t a a c g t g a g t t t t c g t t c c a c t g a g c g t c a g a c c c c g t a g a a a a g a t c a a a g g a t c g g g a a t t g c a c t c a a a a g c a a g g t g a c t c g c a g t c t g g g g c a t c t t t t c t a g t t t c c t a g

7380 ori t t c t t g a g a t c c t t t t t t t c t g c g c g t a a t c t g c t g c t t g c a a a c a a a a a a a c c a c c g c t a a g a a c t c t a g g a a a a a a a g a c g c g c a t t a g a c g a c g a a c g t t t g t t t t t t t g g t g g c g a

7440 ori a c c a g c g g t g g t t t g t t t g c c g g a t c a a g a g c t a c c a a c t c t t t t t c c g a a g g t a a c t g g t g g t c g c c a c c a a a c a a a c g g c c t a g t t c t c g a t g g t t g a g a a a a a g g c t t c c a t t g a c c

7500 ori c t t c a g c a g a g c g c a g a t a c c a a a t a c t g t t c t t c t a g t g t a g c c g t a g t t a g g c c a c c a g a a g t c g t c t c g c g t c t a t g g t t t a t g a c a a g a a g a t c a c a t c g g c a t c a a t c c g g t g g t

AlwNI 7560 ori c t t c a a g a a c t c t g t a g c a c c g c c t a c a t a c c t c g c t c t g c t a a t c c t g t t a c c a g t g g c g a a g t t c t t g a g a c a t c g t g g c g g a t g t a t g g a g c g a g a c g a t t a g g a c a a t g g t c a c c g

7620 ori t g c t g c c a g t g g c g a t a a g t c g t g t c t t a c c g g g t t g g a c t c a a g a c g a t a g t t a c c g g a a c g a c g g t c a c c g c t a t t c a g c a c a g a a t g g c c c a a c c t g a g t t c t g c t a t c a a t g g c c t

7680 ori t a a g g c g c a g c g g t c g g g c t g a a c g g g g g g t t c g t g c a c a c a g c c c a g c t t g g a g c g a a c a t t c c g c g t c g c c a g c c c g a c t t g c c c c c c a a g c a c g t g t g t c g g g t c g a a c c t c g c t t g

7740 ori g a c c t a c a c c g a a c t g a g a t a c c t a c a g c g t g a g c t a t g a g a a a g c g c c a c g c t t c c c g a c t g g a t g t g g c t t g a c t c t a t g g a t g t c g c a c t c g a t a c t c t t t c g c g g t g c g a a g g g c t

7800 ori a g g g a g a a a g g c g g a c a g g t a t c c g g t a a g c g g c a g g g t c g g a a c a g g a g a g c g c a c g a g t c c c t c t t t c c g c c t g t c c a t a g g c c a t t c g c c g t c c c a g c c t t g t c c t c t c g c g t g c t c

7860 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F g g a g c t t c c a g g g g g a a a c g c c t g g t a t c t t t a t a g t c c t g t c g g g t t t c g c c a c c t c t g c c t c g a a g g t c c c c c t t t g c g g a c c a t a g a a a t a t c a g g a c a g c c c a a a g c g g t g g a g a c

7920 ori a c t t g a g c g t c g a t t t t t g t g a t g c t c g t c a g g g g g g c g g a g c c t a t g g a a a a a c g c c a g t g a a c t c g c a g c t a a a a a c a c t a c g a g c a g t c c c c c c g c c t c g g a t a c c t t t t t g c g g t c

7980 c a a c g c g g c c t t t t t a c g g t t c c t g g c c t t t t g c t g g c c t t t t g c t c a c a t g t t c t t t c c g t t g c g c c g g a a a a a t g c c a a g g a c c g g a a a a c g a c c g g a a a a c g a g t g t a c a a g a a a g g

8040 t g c g t t a t c c c c t g a t t c t g t g g a t a a c c g t a t t a c c g c c t t t g a g t g a g c t g a t a c c g c a c g c a a t a g g g g a c t a a g a c a c c t a t t g g c a t a a t g g c g g a a a c t c a c t c g a c t a t g g c g

BspQISapI 8100 A G C G A G T C A G T G A G C G A G L4440 t c g c c g c a g c c g a a c g a c c g a g c g c a g c g a g t c a g t g a g c g a g g a a g c g g a a g a g c g c c c a g c g g c g t c g g c t t g c t g g c t c g c g t c g c t c a g t c a c t c g c t c c t t c g c c t t c t c g c g g g

8160 a a t a c g c a a a c c g c c t c t c c c c g c g c g t t g g c c g a t t c a t t a a t g c a g c t g g c a c g a c a g t t a t g c g t t t g g c g g a g a g g g g c g c g c a a c c g g c t a a g t a a t t a c g t c g a c c g t g c t g t c

8220 CAP binding site g t t t c c c g a c t g g a a a g c g g g c a g t g a g c g c a a c g c a a t t a a t g t g a g t t a g c t c a c t c a c a a a g g g c t g a c c t t t c g c c c g t c a c t c g c g t t g c g t t a a t t a c a c t c a a t c g a g t g a g t

8280 -35 -10 CAP binding site lac promoter lac operator A G M13/pUC Reverse t t a g g c a c c c c a g g c t t t a c a c t t t a t g c t t c c g g c t c g t a t g t t g t g t g g a a t t g t g a g a a t c c g t g g g g t c c g a a a t g t g a a a t a c g a a g g c c g a g c a t a c a a c a c a c c t t a a c a c t c

3ʹ 5ʹ 8323 lac operator M13 rev C G G A T A A C A A T T T C A C A C A G G M13/pUC Reverse C A G G A A A C A G C T A T G A C M13 Reverse c g g a t a a c a a t t t c a c a c a g g a a a c a g c t a t g a c c a t g a t t a c g c c t a t t g t t a a a g t g t g t c c t t t g t c g a t a c t g g t a c t a a t g

Restriction Enzymes

Instructions: By default, all cutters are shown. Filter on number of cut sites or search by enzyme name.

Filter

Features

Primers

BLAST

BLAST (Basic Local Alignment Search Tool) finds regions of similarity between biological sequences. Click on the buttons below to submit a BLAST search to NCBI. The results will appear in a new window. See your recent BLAST results on NCBI's website.

  • Nucleotide-Nucleotide BLAST (BLASTN)

  • Translated Nucleotide-Protein BLAST (BLASTX)

  • Sequence alignment using BLAST (BLAST2)

Sequence Analyzer Guide

Map

Displays a graphical map based on nucleotide sequence data labeled with restriction enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site)

To select a portion of sequence, click one location on the plasmid and then a second location to display the sequence between the two locations.

Sequence

Displays both strands of base paired nucleotide sequences with annotated enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site).

To select a portion of sequence, click one location on the sequence and then a second location to display the sequence between the two locations.

Enzymes

List of restriction enzymes that can cut a given nucleotide sequence. Table lists enzyme name and the sequence location of the cut.

Features

List of common features detected in a given nucleotide sequence. Table lists feature name, location, size, color used to indicate its position on the map, and direction (if relevant).

Primers

List of commonly used primers detected in a given nucleotide sequence. Table lists primer name, sequence, length, binding site location, and direction.

BLAST

Use Basic Local Alignment Search Tool (BLAST) via the NCBI website to determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2.

File Downloads

GenBank

File contains the nucleotide sequence and annotated features in GenBank flat file format. Open the file with a text editor or plasmid mapping software to view the sequence.

SnapGene

File contains the nucleotide sequence and enhanced annotations from SnapGene Server. Open the file with SnapGene software or the free Viewer to view the plasmid map, sequence, and perform additional sequence analysis.