Addgene B-SpCas9-HF1 - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: B-SpCas9-HF1


Map View

Acc65I (6) pBR322ori-F (7897 .. 7916) SacII (7099) FspI (6941) PvuI (6795) Amp-R (6595 .. 6614) pBRforEco (6186 .. 6204) pGEX 3' (6126 .. 6148) pRS-marker (6007 .. 6026) PsiI (5893) F1ori-F (5832 .. 5853) F1ori-R (5622 .. 5641) PluTI (5465) SfoI (5463) NarI (5462) KasI (5461) SbfI (5458) NotI (5313) BGH-rev (5098 .. 5115) SacI (5087) Eco53kI (5085) EcoRI (5074) FseI (5051) nucleoplasmin NLS BsmI (4559) BspQI - SapI (4531) BsaBI * (4417) HindIII (4384) BamHI (4296) KpnI (10) CMV enhancer NdeI (167) SnaBI (273) BseRI (684) PaqCI (789) AgeI (811) SV40 NLS BglII (1190) BmgBI (1479) PspOMI (2295) ApaI (2299) PasI (2709) EcoRV (2793) NruI (3272) PmlI (3729) AflIII - MluI (3896) B-SpCas9-HF1 8056 bp

Sequence View

Acc65IKpnI 60 CMV enhancer c t a g a g g t a c c c g t t a c a t a a c t t a c g g t a a a t g g c c c g c c t g g c t g a c c g c c c a a c g a c g a t c t c c a t g g g c a a t g t a t t g a a t g c c a t t t a c c g g g c g g a c c g a c t g g c g g g t t g c t g

120 CMV enhancer c c c c g c c c a t t g a c g t c a a t a g t a a c g c c a a t a g g g a c t t t c c a t t g a c g t c a a t g g g t g g g g g c g g g t a a c t g c a g t t a t c a t t g c g g t t a t c c c t g a a a g g t a a c t g c a g t t a c c c a c

NdeI 180 CMV enhancer g a g t a t t t a c g g t a a a c t g c c c a c t t g g c a g t a c a t c a a g t g t a t c a t a t g c c a a g t a c g c t c a t a a a t g c c a t t t g a c g g g t g a a c c g t c a t g t a g t t c a c a t a g t a t a c g g t t c a t g c

240 CMV enhancer c c c c c t a t t g a c g t c a a t g a c g g t a a a t g g c c c g c c t g g c a t t g t g c c c a g t a c a t g a c c g g g g g a t a a c t g c a g t t a c t g c c a t t t a c c g g g c g g a c c g t a a c a c g g g t c a t g t a c t g g

SnaBI 300 CMV enhancer chicken β- actin promoter t t a t g g g a c t t t c c t a c t t g g c a g t a c a t c t a c g t a t t a g t c a t c g c t a t t a c c a t g g t c a a t a c c c t g a a a g g a t g a a c c g t c a t g t a g a t g c a t a a t c a g t a g c g a t a a t g g t a c c a g

360 chicken β- actin promoter g a g g t g a g c c c c a c g t t c t g c t t c a c t c t c c c c a t c t c c c c c c c c t c c c c a c c c c c a a t t c t c c a c t c g g g g t g c a a g a c g a a g t g a g a g g g g t a g a g g g g g g g g a g g g g t g g g g g t t a a

420 chicken β- actin promoter t t g t a t t t a t t t a t t t t t t a a t t a t t t t g t g c a g c g a t g g g g g c g g g g g g g g g g g g g g g g a a c a t a a a t a a a t a a a a a a t t a a t a a a a c a c g t c g c t a c c c c c g c c c c c c c c c c c c c c c c

480 chicken β- actin promoter c g c g c g c c a g g c g g g g c g g g g c g g g g c g a g g g g c g g g g c g g g g c g a g g c g g a g a g g t g c g g c g c g c g g t c c g c c c c g c c c c g c c c c g c t c c c c g c c c c g c c c c g c t c c g c c t c t c c a c g c

540 chicken β- actin promoter g c g g c a g c c a a t c a g a g c g g c g c g c t c c g a a a g t t t c c t t t t a t g g c g a g g c g g c g g c g g c g c c g t c g g t t a g t c t c g c c g c g c g a g g c t t t c a a a g g a a a a t a c c g c t c c g c c g c c g c c

600 chicken β- actin promoter hybrid intron c g g c g g c c c t a t a a a a a g c g a a g c g c g c g g c g g g c g g g a g t c g c t g c g a c g c t g c c t t c g g c c g c c g g g a t a t t t t t c g c t t c g c g c g c c g c c c g c c c t c a g c g a c g c t g c g a c g g a a g c

660 hybrid intron c c c c g t g c c c c g c t c c g c c g c c g c c t c g c g c c g c c c g c c c c g g c t c t g a c t g a c c g c g t t g g g g c a c g g g g c g a g g c g g c g g c g g a g c g c g g c g g g c g g g g c c g a g a c t g a c t g g c g c a a

BseRI 720 hybrid intron a c t c c c a c a g g t g a g c g g g c g g g a c g g c c c t t c t c c t c c g g g c t g t a a t t a g c t g a g c a a t g a g g g t g t c c a c t c g c c c g c c c t g c c g g g a a g a g g a g g c c c g a c a t t a a t c g a c t c g t t

780 hybrid intron g a g g t a a g g g t t t a a g g g a t g g t t g g t t g g t g g g g t a t t a a t g t t t a a t t a c c t g g a g c a c t c c a t t c c c a a a t t c c c t a c c a a c c a a c c a c c c c a t a a t t a c a a a t t a a t g g a c c t c g t

AgeIPaqCI 840 1 5 DYKDH hybrid intron 3xFLAG Kozak sequence MDYKDH *TPGTGGHVILVV c c t g c c t g a a a t c a c t t t t t t t c a g g t t g g a c c g g t g c c a c c a t g g a c t a t a a g g a c c a c g g a c g g a c t t t a g t g a a a a a a a g t c c a a c c t g g c c a c g g t g g t a c c t g a t a t t c c t g g t g

900 10 15 20 DGDYKDHDIDYKDDDDK MA P 3xFLAG SV40 NLS DGDYKDHDIDYKDDDDKMAP VSVVLIMINIVFVIVILHGW g a c g g a g a c t a c a a g g a t c a t g a t a t t g a t t a c a a a g a c g a t g a c g a t a a g a t g g c c c c a c t g c c t c t g a t g t t c c t a g t a c t a t a a c t a a t g t t t c t g c t a c t g c t a t t c t a c c g g g g t

960 130 KKKRKV (in frame with SV40 NLS) GIHGVPAA 5 DKKYSI SV40 NLS Cas9 KKKRKVGIHGVPAADKKYSI LLLPLDTDVSDWCGVLLVAD a a g a a g a a g c g g a a g g t c g g t a t c c a c g g a g t c c c a g c a g c c g a c a a g a a g t a c a g c a t c t t c t t c t t c g c c t t c c a g c c a t a g g t g c c t c a g g g t c g t c g g c t g t t c t t c a t g t c g t a g

1020 10 15 20 25 GLDIGTNSVGWAVITDEYKV Cas9 GLDIGTNSVGWAVITDEYKV AQVDAGVRHAPGHDGVLVLH g g c c t g g a c a t c g g c a c c a a c t c t g t g g g c t g g g c c g t g a t c a c c g a c g a g t a c a a g g t g c c g g a c c t g t a g c c g t g g t t g a g a c a c c c g a c c c g g c a c t a g t g g c t g c t c a t g t t c c a c

1080 30 35 40 45 PSKKFKVLGNTDRHSIKKNL Cas9 PSKKFKVLGNTDRHSIKKNL GALFELHQAVGVPVADLLVQ c c c a g c a a g a a a t t c a a g g t g c t g g g c a a c a c c g a c c g g c a c a g c a t c a a g a a g a a c c t g g g g t c g t t c t t t a a g t t c c a c g a c c c g t t g t g g c t g g c c g t g t c g t a g t t c t t c t t g g a c

1140 50 55 60 65 IGALLFDSGETAEATRLKRT Cas9 IGALLFDSGETAEATRLKRT DSGQQEVAAFCGLGGPQLSG a t c g g a g c c c t g c t g t t c g a c a g c g g c g a a a c a g c c g a g g c c a c c c g g c t g a a g a g a a c c t a g c c t c g g g a c g a c a a g c t g t c g c c g c t t t g t c g g c t c c g g t g g g c c g a c t t c t c t t g g

BglII 1200 70 75 80 85 ARRRYTRRKNRICYLQEIFS Cas9 ARRRYTRRKNRICYLQEIFS GSSSVGSPLVPDAIQLLDEA g c c a g a a g a a g a t a c a c c a g a c g g a a g a a c c g g a t c t g c t a t c t g c a a g a g a t c t t c a g c c g g t c t t c t t c t a t g t g g t c t g c c t t c t t g g c c t a g a c g a t a g a c g t t c t c t a g a a g t c g

1260 90 95 100 105 NEMAKVDDSFFHRLEESFLV Cas9 NEMAKVDDSFFHRLEESFLV VLHGLHVVAEEVSQFLGEQH a a c g a g a t g g c c a a g g t g g a c g a c a g c t t c t t c c a c a g a c t g g a a g a g t c c t t c c t g g t g t t g c t c t a c c g g t t c c a c c t g c t g t c g a a g a a g g t g t c t g a c c t t c t c a g g a a g g a c c a c

1320 110 115 120 125 EEDKKHERHPIFGNIVDEVA Cas9 EEDKKHERHPIFGNIVDEVA FLILLVLPVGDEAVDHVLHG g a a g a g g a t a a g a a g c a c g a g c g g c a c c c c a t c t t c g g c a a c a t c g t g g a c g a g g t g g c c c t t c t c c t a t t c t t c g t g c t c g c c g t g g g g t a g a a g c c g t t g t a g c a c c t g c t c c a c c g g

1380 130 135 140 145 YHEKYPTIYHLRKKLVDSTD Cas9 YHEKYPTIYHLRKKLVDSTD VVLLVGGDVVQSLFQHVAGV t a c c a c g a g a a g t a c c c c a c c a t c t a c c a c c t g a g a a a g a a a c t g g t g g a c a g c a c c g a c a t g g t g c t c t t c a t g g g g t g g t a g a t g g t g g a c t c t t t c t t t g a c c a c c t g t c g t g g c t g

1440 150 155 160 165 KADLRLIYLALAHMIKFRGH Cas9 KADLRLIYLALAHMIKFRGH LGVQPQDIQGQGVHDLEPAV a a g g c c g a c c t g c g g c t g a t c t a t c t g g c c c t g g c c c a c a t g a t c a a g t t c c g g g g c c a c t t c c g g c t g g a c g c c g a c t a g a t a g a c c g g g a c c g g g t g t a c t a g t t c a a g g c c c c g g t g

BmgBI 1500 170 175 180 185 FLIEGDLNPDNSDVDKLFIQ Cas9 FLIEGDLNPDNSDVDKLFIQ EQDLAVQVGVVAVHVLQEDL t t c c t g a t c g a g g g c g a c c t g a a c c c c g a c a a c a g c g a c g t g g a c a a g c t g t t c a t c c a g a a g g a c t a g c t c c c g c t g g a c t t g g g g c t g t t g t c g c t g c a c c t g t t c g a c a a g t a g g t c

1560 190 195 200 205 LVQTYNQLFEENPINASGVD Cas9 LVQTYNQLFEENPINASGVD QHLGVVLQELFVGDVGAAHV c t g g t g c a g a c c t a c a a c c a g c t g t t c g a g g a a a a c c c c a t c a a c g c c a g c g g c g t g g a c g a c c a c g t c t g g a t g t t g g t c g a c a a g c t c c t t t t g g g g t a g t t g c g g t c g c c g c a c c t g

1620 210 215 220 225 AKAILSARLSKSRRLENLIA Cas9 AKAILSARLSKSRRLENLIA GLGDQRGSQALASPQFIQDG g c c a a g g c c a t c c t g t c t g c c a g a c t g a g c a a g a g c a g a c g g c t g g a a a a t c t g a t c g c c c g g t t c c g g t a g g a c a g a c g g t c t g a c t c g t t c t c g t c t g c c g a c c t t t t a g a c t a g c g g

1680 230 235 240 245 QLPGEKKNGLFGNLIALSLG Cas9 QLPGEKKNGLFGNLIALSLG LQGALLLIAQESVQNGQAQA c a g c t g c c c g g c g a g a a g a a g a a t g g c c t g t t c g g a a a c c t g a t t g c c c t g a g c c t g g g c g t c g a c g g g c c g c t c t t c t t c t t a c c g g a c a a g c c t t t g g a c t a a c g g g a c t c g g a c c c g

1740 250 255 260 265 LTPNFKSNFDLAEDAKLQLS Cas9 LTPNFKSNFDLAEDAKLQLS QGGVELAVEVQGLIGFQLQA c t g a c c c c c a a c t t c a a g a g c a a c t t c g a c c t g g c c g a g g a t g c c a a a c t g c a g c t g a g c g a c t g g g g g t t g a a g t t c t c g t t g a a g c t g g a c c g g c t c c t a c g g t t t g a c g t c g a c t c g

1800 270 275 280 285 KDTYDDDLDNLLAQIGDQYA Cas9 KDTYDDDLDNLLAQIGDQYA LVGVVVVQVVQQGLDAVLVG a a g g a c a c c t a c g a c g a c g a c c t g g a c a a c c t g c t g g c c c a g a t c g g c g a c c a g t a c g c c t t c c t g t g g a t g c t g c t g c t g g a c c t g t t g g a c g a c c g g g t c t a g c c g c t g g t c a t g c g g

1860 290 295 300 305 DLFLAAKNLSDAILLSDILR Cas9 DLFLAAKNLSDAILLSDILR VQKQGGLVQGVGDQQAVDQS g a c c t g t t t c t g g c c g c c a a g a a c c t g t c c g a c g c c a t c c t g c t g a g c g a c a t c c t g a g a c t g g a c a a a g a c c g g c g g t t c t t g g a c a g g c t g c g g t a g g a c g a c t c g c t g t a g g a c t c t

1920 310 315 320 325 VNTEITKAPLSASMIKRYDE Cas9 VNTEITKAPLSASMIKRYDE HVGLDGLGGQAGRHDLSVVL g t g a a c a c c g a g a t c a c c a a g g c c c c c c t g a g c g c c t c t a t g a t c a a g a g a t a c g a c g a g c a c t t g t g g c t c t a g t g g t t c c g g g g g g a c t c g c g g a g a t a c t a g t t c t c t a t g c t g c t c

1980 330 335 340 345 HHQDLTLLKALVRQQLPEKY Cas9 HHQDLTLLKALVRQQLPEKY VVLVQGQQFSEHPLLQRLLV c a c c a c c a g g a c c t g a c c c t g c t g a a a g c t c t c g t g c g g c a g c a g c t g c c t g a g a a g t a c g t g g t g g t c c t g g a c t g g g a c g a c t t t c g a g a g c a c g c c g t c g t c g a c g g a c t c t t c a t g

2040 350 355 360 365 KEIFFDQSKNGYAGYIDGGA Cas9 KEIFFDQSKNGYAGYIDGGA FLNEEVLALVAVGAVNVASG a a a g a g a t t t t c t t c g a c c a g a g c a a g a a c g g c t a c g c c g g c t a c a t t g a c g g c g g a g c c t t t c t c t a a a a g a a g c t g g t c t c g t t c t t g c c g a t g c g g c c g a t g t a a c t g c c g c c t c g g

2100 370 375 380 385 SQEEFYKFIKPILEKMDGTE Cas9 SQEEFYKFIKPILEKMDGTE ALFLEVLEDLGDQFLHVAGL a g c c a g g a a g a g t t c t a c a a g t t c a t c a a g c c c a t c c t g g a a a a g a t g g a c g g c a c c g a g t c g g t c c t t c t c a a g a t g t t c a a g t a g t t c g g g t a g g a c c t t t t c t a c c t g c c g t g g c t c

2160 390 395 400 405 ELLVKLNREDLLRKQRTFDN Cas9 ELLVKLNREDLLRKQRTFDN FQEHLQVSLVQQPLLPGEVV g a a c t g c t c g t g a a g c t g a a c a g a g a g g a c c t g c t g c g g a a g c a g c g g a c c t t c g a c a a c c t t g a c g a g c a c t t c g a c t t g t c t c t c c t g g a c g a c g c c t t c g t c g c c t g g a a g c t g t t g

2220 410 415 420 425 GSIPHQIHLGELHAILRRQE Cas9 GSIPHQIHLGELHAILRRQE AADGVLDVQSLQVGNQPPLF g g c a g c a t c c c c c a c c a g a t c c a c c t g g g a g a g c t g c a c g c c a t t c t g c g g c g g c a g g a a c c g t c g t a g g g g g t g g t c t a g g t g g a c c c t c t c g a c g t g c g g t a a g a c g c c g c c g t c c t t

2280 430 435 440 445 DFYPFLKDNREKIEKILTFR Cas9 DFYPFLKDNREKIEKILTFR IKVWEQLVVPFLDLLDQGEA g a t t t t t a c c c a t t c c t g a a g g a c a a c c g g g a a a a g a t c g a g a a g a t c c t g a c c t t c c g c c t a a a a a t g g g t a a g g a c t t c c t g t t g g c c c t t t t c t a g c t c t t c t a g g a c t g g a a g g c g

PspOMIApaI 2340 450 455 460 465 IPYYVGPLARGNSRFAWMTR Cas9 IPYYVGPLARGNSRFAWMTR DGVVHARQGPSVASEGPHGS a t c c c c t a c t a c g t g g g c c c t c t g g c c a g g g g a a a c a g c a g a t t c g c c t g g a t g a c c a g a t a g g g g a t g a t g c a c c c g g g a g a c c g g t c c c c t t t g t c g t c t a a g c g g a c c t a c t g g t c t

2400 470 475 480 485 KSEETITPWNFEEVVDKGAS Cas9 KSEETITPWNFEEVVDKGAS LALFGDGGPVELFHHVLASG a a g a g c g a g g a a a c c a t c a c c c c c t g g a a c t t c g a g g a a g t g g t g g a c a a g g g c g c t t c c t t c t c g c t c c t t t g g t a g t g g g g g a c c t t g a a g c t c c t t c a c c a c c t g t t c c c g c g a a g g

2460 490 495 500 505 AQSFIERMTAFDKNLPNEKV Cas9 AQSFIERMTAFDKNLPNEKV GLAEDLPHGGEILVQGVLLH g c c c a g a g c t t c a t c g a g c g g a t g a c c g c c t t c g a t a a g a a c c t g c c c a a c g a g a a g g t g c g g g t c t c g a a g t a g c t c g c c t a c t g g c g g a a g c t a t t c t t g g a c g g g t t g c t c t t c c a c

2520 510 515 520 525 LPKHSLLYEYFTVYNELTKV Cas9 LPKHSLLYEYFTVYNELTKV QGLVAQQVLVEGHIVLQGFH c t g c c c a a g c a c a g c c t g c t g t a c g a g t a c t t c a c c g t g t a t a a c g a g c t g a c c a a a g t g g a c g g g t t c g t g t c g g a c g a c a t g c t c a t g a a g t g g c a c a t a t t g c t c g a c t g g t t t c a c

2580 530 535 540 545 KYVTEGMRKPAFLSGEQKKA Cas9 KYVTEGMRKPAFLSGEQKKA FVHGLSHSLGGEQAALLFLG a a a t a c g t g a c c g a g g g a a t g a g a a a g c c c g c c t t c c t g a g c g g c g a g c a g a a a a a g g c c t t t a t g c a c t g g c t c c c t t a c t c t t t c g g g c g g a a g g a c t c g c c g c t c g t c t t t t t c c g g

2640 550 555 560 565 IVDLLFKTNRKVTVKQLKED Cas9 IVDLLFKTNRKVTVKQLKED DHVQQELGVPFHGHLLQFLV a t c g t g g a c c t g c t g t t c a a g a c c a a c c g g a a a g t g a c c g t g a a g c a g c t g a a a g a g g a c t a g c a c c t g g a c g a c a a g t t c t g g t t g g c c t t t c a c t g g c a c t t c g t c g a c t t t c t c c t g

2700 570 575 580 585 YFKKIECFDSVEISGVEDRF Cas9 YFKKIECFDSVEISGVEDRF VELFDLAEVGHFDGAHFIPE t a c t t c a a g a a a a t c g a g t g c t t c g a c t c c g t g g a a a t c t c c g g c g t g g a a g a t c g g t t c a t g a a g t t c t t t t a g c t c a c g a a g c t g a g g c a c c t t t a g a g g c c g c a c c t t c t a g c c a a g

PasI 2760 590 595 600 605 NASLGTYHDLLKIIKDKDFL Cas9 NASLGTYHDLLKIIKDKDFL VGGQACVVIQQFNDLVLVEQ a a c g c c t c c c t g g g c a c a t a c c a c g a t c t g c t g a a a a t t a t c a a g g a c a a g g a c t t c c t g t t g c g g a g g g a c c c g t g t a t g g t g c t a g a c g a c t t t t a a t a g t t c c t g t t c c t g a a g g a c

EcoRV 2820 610 615 620 625 DNEENEDILEDIVLTLTLFE Cas9 DNEENEDILEDIVLTLTLFE VILFVLVNQFIDHQGQCQKL g a c a a t g a g g a a a a c g a g g a c a t t c t g g a a g a t a t c g t g c t g a c c c t g a c a c t g t t t g a g c t g t t a c t c c t t t t g c t c c t g t a a g a c c t t c t a t a g c a c g a c t g g g a c t g t g a c a a a c t c

2880 630 635 640 645 DREMIEERLKTYAHLFDDKV Cas9 DREMIEERLKTYAHLFDDKV *AWRNSSLT VSLHDLFPQFGIGVQEVVFH g a c a g a g a g a t g a t c g a g g a a c g g c t g a a a a c c t a t g c c c a c c t g t t c g a c g a c a a a g t g c t g t c t c t c t a c t a g c t c c t t g c c g a c t t t t g g a t a c g g g t g g a c a a g c t g c t g t t t c a c

2940 650 655 660 665 MKQLKRRRYTGWGALSRKLI Cas9 MKQLKRRRYTGWGALSRKLI IFCSFRRLYVPQPASLRFSI HLLQLPPSVGAPARQAPLQD a t g a a g c a g c t g a a g c g g c g g a g a t a c a c c g g c t g g g g c g c g c t g a g c c g g a a g c t g a t c t a c t t c g t c g a c t t c g c c g c c t c t a t g t g g c c g a c c c c g c g c g a c t c g g c c t t c g a c t a g

3000 670 675 680 685 NGIRDKQSGKTILDFLKSDG Cas9 NGIRDKQSGKTILDFLKSDG LPMRSLCDPLVIRSKRFDSP VADAVLLGALCDQIEQLGVA a a c g g c a t c c g c g a c a a g c a g t c c g g c a a g a c a a t c c t g g a t t t c c t g a a g t c c g a c g g c t t g c c g t a g g c g c t g t t c g t c a g g c c g t t c t g t t a g g a c c t a a a g g a c t t c a g g c t g c c g

3060 690 695 700 705 FANRNFMALIHDDSLTFKED Cas9 FANRNFMALIHDDSLTFKED KALLFKMASIWSSLRVKLSS EGVSVEHRQDVVVAQGKFLV t t c g c c a a c a g a a a c t t c a t g g c g c t g a t c c a c g a c g a c a g c c t g a c c t t t a a a g a g g a c a a g c g g t t g t c t t t g a a g t a c c g c g a c t a g g t g c t g c t g t c g g a c t g g a a a t t t c t c c t g

3120 710 715 720 725 IQKAQVSGQGDSLHEHIANL Cas9 IQKAQVSGQGDSLHEHIANL MWFAWTDPWPSLRCSCMALR DLFGLHGALAIAQVLVNGIE a t c c a g a a a g c c c a g g t g t c c g g c c a g g g c g a t a g c c t g c a c g a g c a c a t t g c c a a t c t c t a g g t c t t t c g g g t c c a c a g g c c g g t c c c g c t a t c g g a c g t g c t c g t g t a a c g g t t a g a g

3180 730 735 740 745 AGSPAIKKGILQTVKVVDEL Cas9 AGSPAIKKGILQTVKVVDEL APLGAMLFPMRWVTFTTSSS GAAGGNLLADELCHLHHVLQ g c c g g c a g c c c c g c c a t t a a g a a g g g c a t c c t c c a g a c a g t g a a g g t g g t g g a c g a g c t g c g g c c g t c g g g g c g g t a a t t c t t c c c g t a g g a g g t c t g t c a c t t c c a c c a c c t g c t c g a c

3240 750 755 760 765 VKVMGRHKPENIVIEMAREN Cas9 VKVMGRHKPENIVIEMAREN TFTIPRCLGSFMTISIALSF HFHHAPVLGLVDHDFHGSLV g t g a a a g t g a t g g g c c g g c a c a a g c c c g a g a a c a t c g t g a t c g a a a t g g c c a g a g a g a a c c a c t t t c a c t a c c c g g c c g t g t t c g g g c t c t t g t a g c a c t a g c t t t a c c g g t c t c t c t t g

NruI 3300 770 775 780 785 QTTQKGQKNSRERMKRIEEG Cas9 QTTQKGQKNSRERMKRIEEG WVVWFPCFFLRSLIFRISSP LGGLLSLLVTALSHLPDFLA c a g a c c a c c c a g a a g g g a c a g a a g a a c a g t c g c g a g a g a a t g a a g c g g a t c g a a g a g g g c g t c t g g t g g g t c t t c c c t g t c t t c t t g t c a g c g c t c t c t t a c t t c g c c t a g c t t c t c c c g

3360 790 795 800 805 IKELGSQILKEHPVENTQLQ Cas9 IKELGSQILKEHPVENTQLQ M DFLQAALDQFFVGHFVGLEL a t c a a a g a g c t g g g c a g c c a g a t c c t g a a a g a a c a c c c c g t g g a a a a c a c c c a g c t c c a g t a g t t t c t c g a c c c g t c g g t c t a g g a c t t t c t t g t g g g g c a c c t t t t g t g g g t c g a g g t c

3420 810 815 820 825 NEKLYLYYLQNGRDMYVDQE Cas9 NEKLYLYYLQNGRDMYVDQE VLLQVQVVELIPPIHVHVLF a a c g a g a a g c t g t a c c t g t a c t a c c t c c a g a a t g g g c g g g a t a t g t a c g t g g a c c a g g a a t t g c t c t t c g a c a t g g a c a t g a t g g a g g t c t t a c c c g c c c t a t a c a t g c a c c t g g t c c t t

3480 830 835 840 845 LDINRLSDYDVDHIVPQSFL Cas9 LDINRLSDYDVDHIVPQSFL QVDVPQGVVIHVM c t g g a c a t c a a c c g g c t g t c c g a c t a c g a t g t g g a c c a t a t c g t g c c t c a g a g c t t t c t t g a c c t g t a g t t g g c c g a c a g g c t g a t g c t a c a c c t g g t a t a g c a c g g a g t c t c g a a a g a a

3540 850 855 860 865 KDDSIDNKVLTRSDKNRGKS Cas9 KDDSIDNKVLTRSDKNRGKS a a g g a c g a c t c c a t c g a c a a c a a g g t g c t g a c c a g a a g c g a c a a g a a c c g g g g c a a g a g c t t c c t g c t g a g g t a g c t g t t g t t c c a c g a c t g g t c t t c g c t g t t c t t g g c c c c g t t c t c g

3600 870 875 880 885 DNVPSEEVVKKMKNYWRQLL Cas9 DNVPSEEVVKKMKNYWRQLL g a c a a c g t g c c c t c c g a a g a g g t c g t g a a g a a g a t g a a g a a c t a c t g g c g g c a g c t g c t g c t g t t g c a c g g g a g g c t t c t c c a g c a c t t c t t c t a c t t c t t g a t g a c c g c c g t c g a c g a c

3660 890 895 900 905 NAKLITQRKFDNLTKAERGG Cas9 NAKLITQRKFDNLTKAERGG a a c g c c a a g c t g a t t a c c c a g a g a a a g t t c g a c a a t c t g a c c a a g g c c g a g a g a g g c g g c t t g c g g t t c g a c t a a t g g g t c t c t t t c a a g c t g t t a g a c t g g t t c c g g c t c t c t c c g c c g

3720 910 915 920 925 LSELDKAGFIKRQLVETRAI Cas9 LSELDKAGFIKRQLVETRAI c t g a g c g a a c t g g a t a a g g c c g g c t t c a t c a a g a g a c a g c t g g t g g a a a c c c g g g c g a t c g a c t c g c t t g a c c t a t t c c g g c c g a a g t a g t t c t c t g t c g a c c a c c t t t g g g c c c g c t a g

PmlI 3780 930 935 940 945 TKHVAQILDSRMNTKYDEND Cas9 TKHVAQILDSRMNTKYDEND a c a a a g c a c g t g g c a c a g a t c c t g g a c t c c c g g a t g a a c a c t a a g t a c g a c g a g a a t g a c t g t t t c g t g c a c c g t g t c t a g g a c c t g a g g g c c t a c t t g t g a t t c a t g c t g c t c t t a c t g

3840 950 955 960 965 KLIREVKVITLKSKLVSDFR Cas9 KLIREVKVITLKSKLVSDFR a a g c t g a t c c g g g a a g t g a a a g t g a t c a c c c t g a a g t c c a a g c t g g t g t c c g a t t t c c g g t t c g a c t a g g c c c t t c a c t t t c a c t a g t g g g a c t t c a g g t t c g a c c a c a g g c t a a a g g c c

AflIIIMluI 3900 970 975 980 985 KDFQFYKVREINNYHHAHDA Cas9 KDFQFYKVREINNYHHAHDA a a g g a t t t c c a g t t t t a c a a a g t g c g c g a g a t c a a c a a c t a c c a c c a c g c c c a c g a c g c g t t c c t a a a g g t c a a a a t g t t t c a c g c g c t c t a g t t g t t g a t g g t g g t g c g g g t g c t g c g c

3960 990 995 1000 1005 YLNAVVGTALIKKYPKLGGK Cas9 YLNAVVGTALIKKYPKLGGK t a c c t g a a c g c c g t c g t g g g a a c c g c c c t g a t c a a a a a g t a c c c t a a g c t g g g t g g c a a g a t g g a c t t g c g g c a g c a c c c t t g g c g g g a c t a g t t t t t c a t g g g a t t c g a c c c a c c g t t c

4020 1010 1015 1020 1025 VYDVRKMIAKSEQEIGKATA Cas9 VYDVRKMIAKSEQEIGKATA g t g t a c g a t g t a c g g a a g a t g a t c g c c a a g a g c g a g c a g g a a a t c g g c a a g g c t a c c g c c c a c a t g c t a c a t g c c t t c t a c t a g c g g t t c t c g c t c g t c c t t t a g c c g t t c c g a t g g c g g

4080 1030 1035 1040 1045 KYFFYSNIMNFFKTEITLAN Cas9 KYFFYSNIMNFFKTEITLAN a a g t a c t t c t t c t a c a g c a a c a t c a t g a a c t t t t t c a a g a c c g a g a t t a c c c t g g c c a a c t t c a t g a a g a a g a t g t c g t t g t a g t a c t t g a a a a a g t t c t g g c t c t a a t g g g a c c g g t t g

4140 1050 1055 1060 1065 GEIRKRPLIETNGETGEIVW Cas9 GEIRKRPLIETNGETGEIVW g g c g a g a t c c g g a a g c g g c c t c t g a t c g a g a c a a a c g g c g a a a c c g g g g a g a t c g t g t g g c c g c t c t a g g c c t t c g c c g g a g a c t a g c t c t g t t t g c c g c t t t g g c c c c t c t a g c a c a c c

4200 1070 1075 1080 1085 DKGRDFATVRKVLSMPQVNI Cas9 DKGRDFATVRKVLSMPQVNI g a t a a g g g c c g g g a t t t t g c c a c c g t g c g g a a a g t g c t g a g c a t g c c c c a a g t g a a t a t c c t a t t c c c g g c c c t a a a a c g g t g g c a c g c c t t t c a c g a c t c g t a c g g g g t t c a c t t a t a g

4260 1090 1095 1100 1105 VKKTEVQTGGFSKESILPKR Cas9 VKKTEVQTGGFSKESILPKR g t g a a a a a g a c c g a g g t g c a g a c a g g c g g c t t c a g c a a a g a g t c t a t c c t g c c c a a g a g g c a c t t t t t c t g g c t c c a c g t c t g t c c g c c g a a g t c g t t t c t c a g a t a g g a c g g g t t c t c c

BamHI 4320 1110 1115 1120 1125 NSDKLIARKKDWDPKKYGGF Cas9 NSDKLIARKKDWDPKKYGGF a a c a g c g a t a a g c t g a t c g c c a g a a a g a a g g a c t g g g a t c c t a a g a a g t a c g g c g g c t t c t t g t c g c t a t t c g a c t a g c g g t c t t t c t t c c t g a c c c t a g g a t t c t t c a t g c c g c c g a a g

4380 1130 1135 1140 1145 DSPTVAYSVLVVAKVEKGKS Cas9 DSPTVAYSVLVVAKVEKGKS g a c a g c c c c a c c g t g g c c t a t t c t g t g c t g g t g g t g g c c a a a g t g g a a a a g g g c a a g t c c c t g t c g g g g t g g c a c c g g a t a a g a c a c g a c c a c c a c c g g t t t c a c c t t t t c c c g t t c a g g

BsaBI*HindIII 4440 1150 1155 1160 1165 KKLKSVKELLGITIMERSSF Cas9 KKLKSVKELLGITIMERSSF a a g a a g c t t a a g a g t g t g a a a g a g c t g c t g g g g a t c a c c a t c a t g g a a a g a a g c a g c t t c t t c t t c g a a t t c t c a c a c t t t c t c g a c g a c c c c t a g t g g t a g t a c c t t t c t t c g t c g a a g

4500 1170 1175 1180 1185 EKNPIDFLEAKGYKEVKKDL Cas9 EKNPIDFLEAKGYKEVKKDL g a g a a g a a t c c c a t c g a c t t t c t g g a a g c c a a g g g c t a c a a a g a a g t g a a a a a g g a c c t g c t c t t c t t a g g g t a g c t g a a a g a c c t t c g g t t c c c g a t g t t t c t t c a c t t t t t c c t g g a c

BsmIBspQISapI 4560 1190 1195 1200 1205 IIKLPKYSLFELENGRKRML Cas9 IIKLPKYSLFELENGRKRML a t c a t c a a g c t g c c t a a g t a c t c g c t c t t c g a g c t g g a a a a c g g c c g g a a g a g a a t g c t g t a g t a g t t c g a c g g a t t c a t g a g c g a g a a g c t c g a c c t t t t g c c g g c c t t c t c t t a c g a c

4620 1210 1215 1220 1225 ASAGELQKGNELALPSKYVN Cas9 ASAGELQKGNELALPSKYVN g c c t c t g c c g g c g a a c t g c a g a a g g g a a a c g a a c t g g c c c t g c c c t c c a a a t a t g t g a a c c g g a g a c g g c c g c t t g a c g t c t t c c c t t t g c t t g a c c g g g a c g g g a g g t t t a t a c a c t t g

4680 1230 1235 1240 1245 FLYLASHYEKLKGSPEDNEQ Cas9 FLYLASHYEKLKGSPEDNEQ t t c c t g t a c c t g g c c a g c c a c t a t g a g a a g c t g a a g g g c t c c c c c g a g g a t a a t g a g c a g a a g g a c a t g g a c c g g t c g g t g a t a c t c t t c g a c t t c c c g a g g g g g c t c c t a t t a c t c g t c

4740 1250 1255 1260 1265 KQLFVEQHKHYLDEIIEQIS Cas9 KQLFVEQHKHYLDEIIEQIS a a a c a g c t g t t t g t g g a a c a g c a c a a g c a c t a c c t g g a c g a g a t c a t c g a g c a g a t c a g c t t t g t c g a c a a a c a c c t t g t c g t g t t c g t g a t g g a c c t g c t c t a g t a g c t c g t c t a g t c g

4800 1270 1275 1280 1285 EFSKRVILADANLDKVLSAY Cas9 EFSKRVILADANLDKVLSAY g a g t t c t c c a a g a g a g t g a t c c t g g c c g a c g c t a a t c t g g a c a a a g t g c t g t c c g c c t a c c t c a a g a g g t t c t c t c a c t a g g a c c g g c t g c g a t t a g a c c t g t t t c a c g a c a g g c g g a t g

4860 1290 1295 1300 1305 NKHRDKPIREQAENIIHLFT Cas9 NKHRDKPIREQAENIIHLFT a a c a a g c a c c g g g a t a a g c c c a t c a g a g a g c a g g c c g a g a a t a t c a t c c a c c t g t t t a c c t t g t t c g t g g c c c t a t t c g g g t a g t c t c t c g t c c g g c t c t t a t a g t a g g t g g a c a a a t g g

4920 1310 1315 1320 1325 LTNLGAPAAFKYFDTTIDRK Cas9 LTNLGAPAAFKYFDTTIDRK c t g a c c a a t c t g g g a g c c c c t g c c g c c t t c a a g t a c t t t g a c a c c a c c a t c g a c c g g a a g g a c t g g t t a g a c c c t c g g g g a c g g c g g a a g t t c a t g a a a c t g t g g t g g t a g c t g g c c t t c

4980 1330 1335 1340 1345 RYTSTKEVLDATLIHQSITG Cas9 RYTSTKEVLDATLIHQSITG a g g t a c a c c a g c a c c a a a g a g g t g c t g g a c g c c a c c c t g a t c c a c c a g a g c a t c a c c g g c t c c a t g t g g t c g t g g t t t c t c c a c g a c c t g c g g t g g g a c t a g g t g g t c t c g t a g t g g c c g

5040 1350 1355 1360 LYETRIDLSQLGGD 1 5 KRPAAT Cas9 nucleoplasmin NLS LYETRIDLSQLGGDKRPAAT c t g t a c g a g a c a c g g a t c g a c c t g t c t c a g c t g g g a g g c g a c a a a a g g c c g g c g g c c a c g g a c a t g c t c t g t g c c t a g c t g g a c a g a g t c g a c c c t c c g c t g t t t t c c g g c c g c c g g t g c

Eco53kISacIEcoRIFseI 5100 10 15 KKAGQAKKKK * nucleoplasmin NLS G G A BGH-rev KKAGQAKKKK* a a a a a g g c c g g c c a g g c a a a a a a g a a a a a g t a a g a a t t c c t a g a g c t c g c t g a t c a g c c t t t t t t c c g g c c g g t c c g t t t t t t c t t t t t c a t t c t t a a g g a t c t c g a g c g a c t a g t c g g a

5160 bGH poly(A) signal G C T G A C A C G G A A G A T BGH-rev c g a c t g t g c c t t c t a g t t g c c a g c c a t c t g t t g t t t g c c c c t c c c c c g t g c c t t c c t t g a g c t g a c a c g g a a g a t c a a c g g t c g g t a g a c a a c a a a c g g g g a g g g g g c a c g g a a g g a a c t

5220 bGH poly(A) signal c c c t g g a a g g t g c c a c t c c c a c t g t c c t t t c c t a a t a a a a t g a g g a a a t t g c a t c g c a t t g g g a c c t t c c a c g g t g a g g g t g a c a g g a a a g g a t t a t t t t a c t c c t t t a a c g t a g c g t a a

5280 bGH poly(A) signal g t c t g a g t a g g t g t c a t t c t a t t c t g g g g g g t g g g g t g g g g c a g g a c a g c a a g g g g g a g g c a g a c t c a t c c a c a g t a a g a t a a g a c c c c c c a c c c c a c c c c g t c c t g t c g t t c c c c c t c c

NotI 5340 bGH poly(A) signal AAV2 ITR AAV2 ITR a t t g g g a a g a g a a t a g c a g g c a t g c t g g g g a g c g g c c g c a g g a a c c c c t a g t g a t g g a g t t a a c c c t t c t c t t a t c g t c c g t a c g a c c c c t c g c c g g c g t c c t t g g g g a t c a c t a c c t c a

5400 AAV2 ITR AAV2 ITR t g g c c a c t c c c t c t c t g c g c g c t c g c t c g c t c a c t g a g g c c g g g c g a c c a a a g g t c g c c c a c c g g t g a g g g a g a g a c g c g c g a g c g a g c g a g t g a c t c c g g c c c g c t g g t t t c c a g c g g g

SbfI 5460 AAV2 ITR AAV2 ITR g a c g c c c g g g c t t t g c c c g g g c g g c c t c a g t g a g c g a g c g a g c g c g c a g c t g c c t g c a g g c t g c g g g c c c g a a a c g g g c c c g c c g g a g t c a c t c g c t c g c t c g c g c g t c g a c g g a c g t c c

KasINarISfoIPluTI 5520 g g c g c c t g a t g c g g t a t t t t c t c c 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 c c g c g g a c t a c g c c a t a a a a g a g 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

5580 f1 ori a a a g c a a c c a t a g t a c g c g c c c t g 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 t t t c g t t g g t a t c a t g c g c g g g a 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

5640 f1 ori G A G G A A A G C G A A A G A A G G G F1ori-R g c g c a g c g t g a c c g c t a c a c t t g c 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 c g c g t c g c a c t g g c g a t g t g a a c 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

5700 f1 ori A F1ori-R t t c c t t t c t c g c c a c g t t c g c c g g 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 a g g a a a g a g c g g t g c a a g c g g c c 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

5760 f1 ori 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 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 c c c a a g g c t a a a t c a c g a a a t g c 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

5820 f1 ori t t c a c g t a g t g g g c c a t c g c c c t g 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 a a g t g c a t c a c c c g g t a g c g g g a c 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

5880 f1 ori G T G G A C T C T T G T T C C A A A C T G G F1ori-F g t t c t t t a a t a g t g g a c t c t t g t t 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 c a a g a a a t t a t c a c c t g a g a a c a a 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

PsiI 5940 f1 ori 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 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 a a g a a a a c t a a a t a t t c c c t a a a a 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

6000 f1 ori t t a a c a a a a a t t t a a c g c g a a t t t 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 a a t t g t t t t t a a a t t g c g c t t a a a 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

6060 A T G T T A G A C G A G A C T A C G G C pRS-marker t c t c a g t a c a a t c t g c t c t g a t g c 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 a g a g t c a t g t t a g a c g a g a c t a c 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

6120 c g c t g a c g c g c c c t g a c g g g c t t g 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 g c g a c t g c g c g g g a c t g c c c g a a c 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

6180 C C G G G A G C T G C A T G T G T C A G A G G pGEX 3' c g t c t c c g g g a g c t g c a t g t g t c a 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 g c a g a g g c c c t c g a c g t a c a c a g 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

6240 C G G A G C A C T A T G C G G A T A A pBRforEco a a a g g g c c t c g t g a t a c g c c t a t t 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 t t t c c c g g a g c a c t a t g c g g a t a a 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

6300 AmpR promoter 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 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 c t g c a g t c c a c c g t g a a a a g c c c c 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

6360 AmpR promoter a a t a c a t t c a a a t a t g t a t c c g c t 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 a t g t a a g t t t a t a c a t a g g c g a 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

6420 1 5 10 15 signal sequence MSIQHFRVALIPFFA AmpR promoter AmpR MSIQHFRVALIPFFA t t g a a a a a g g a a g a g t a t g a g t a t 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 a a c t t t t t c c t t c t c a t a c t c a t a 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

6480 20 signal sequence AFCLPVFA 25 30 35 HPETLVKVKDAE AmpR AFCLPVFAHPETLVKVKDAE g g c a t t t t g c c t t c c t g t t t t t g c 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 c c g t a a a a c g g a a g g a c a a a a a c 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

6540 40 45 50 55 DQLGARVGYIELDLNSGKIL AmpR DQLGARVGYIELDLNSGKIL a g a t c a g t t g g g t g c a c g a g t g g g 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 c t a g t c a a c c c a c g t g c t c a c c c 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

6600 60 65 70 75 ESFRPEERFPMMSTFKVLLC AmpR C G A T A C Amp-R ESFRPEERFPMMSTFKVLLC t g a g a g t t t t c g c c c c g a a g a a c g 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 a c t c t c a a a a g c g g g g c t t c t t g c 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

6660 80 85 90 95 GAVLSRIDAGQEQLGRRIHY AmpR A C C G C G C C A T A A T A Amp-R GAVLSRIDAGQEQLGRRIHY t g g c g c g g t a t t a t c c c g t a t t g a 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 a c c g c g c c a t a a t a g g g c a t a a c t 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

6720 100 105 110 115 SQNDLVEYSPVTEKHLTDGM AmpR SQNDLVEYSPVTEKHLTDGM t t c t c a g a a t g a c t t g g t t g a g t a 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 a a g a g t c t t a c t g a a c c a a c t c a t 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

6780 120 125 130 135 TVRELCSAAITMSDNTAANL AmpR TVRELCSAAITMSDNTAANL g a c a g t a a g a g a a t t a t g c a g t g c 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 c t g t c a t t c t c t t a a t a c g t c a c g 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

PvuI 6840 140 145 150 155 LLTTIGGPKELTAFLHNMGD AmpR LLTTIGGPKELTAFLHNMGD a c t t c t g a c a a c g a t c g g a g g a c c 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 g a a g a c t g t t g c t a g c c t c c t g 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

6900 160 165 170 175 HVTRLDRWEPELNEAIPNDE AmpR HVTRLDRWEPELNEAIPNDE t c a t g t a a c t c g c c t t g a t c g t t g 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 a g t a c a t t g a g c g g a a c t a g c a a 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

FspI 6960 180 185 190 195 RDTTMPVAMATTLRKLLTGE AmpR RDTTMPVAMATTLRKLLTGE g c g t g a c a c c a c g a t g c c t g t a g c 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 c g c a c t g t g g t g c t a c g g a c a t c g 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

7020 200 205 210 215 LLTLASRQQLIDWMEADKVA AmpR LLTLASRQQLIDWMEADKVA a c t a c t t a c t c t a g c t t c c c g g c a 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 t g a t g a a t g a g a t c g a a g g g c c g 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

7080 220 225 230 235 GPLLRSALPAGWFIADKSGA AmpR GPLLRSALPAGWFIADKSGA 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 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 t c c t g g t g a a g a c g c g a g c c g g g a 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

SacII 7140 240 245 250 255 GERGSRGIIAALGPDGKPSR AmpR GERGSRGIIAALGPDGKPSR c g g t g a g c g t g g a a g c c g c g g t a t 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 g c c a c t c g c a c c t t c g g c g c c a t a 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

7200 260 265 270 275 IVVIYTTGSQATMDERNRQI AmpR IVVIYTTGSQATMDERNRQI 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 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 a t a g c a t c a a t a g a t g t g c t g c c c 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

7260 280 285 AEIGASLIKHW* AmpR AEIGASLIKHW* c g c t g a g a t a g g t g c c t c a c t g a t 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 g c g a c t c t a t c c a c g g a g t g a c t a 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

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

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

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

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

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

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

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

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

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

7860 ori a t g a g a a a g c g c c a c g c t t c c c g a 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 t a c t c t t t c g c g g t g c g a a g g g c t 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

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

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

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

8056 g c c t t t t g c t c a c a t g c g g a a a a c g a g t g t a c

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.