Addgene pISL1.1.0-gDNA Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pISL1.1.0-gDNA Sequencing Result


Map View

LKO.1 5' (8361 .. 8380) hU6-F (8190 .. 8210) AflIII - PciI (8184) pBR322ori-F (8029 .. 8048) SacII (7231) FspI (7073) PvuI (6927) Amp-R (6727 .. 6746) pBRforEco (6318 .. 6336) pGEX 3' (6258 .. 6280) pRS-marker (6139 .. 6158) PsiI (6025) F1ori-F (5964 .. 5985) F1ori-R (5754 .. 5773) PluTI (5597) SfoI (5595) NarI (5594) KasI (5593) SbfI (5590) NotI (5445) BGH-rev (5230 .. 5247) EcoRI (5206) FseI (5183) nucleoplasmin NLS BsmI (4691) BsaBI * (4549) XbaI (157) Acc65I (163) KpnI (167) SnaBI (430) BseRI (795) PaqCI (900) AgeI (922) BglII (1301) PspOMI (2406) ApaI (2410) PasI (2820) EcoRV (2904) PmlI (3840) KflI (4428) pISL1.1.0-gDNA 8463 bp

Sequence View

5ʹ 3ʹ 60 gRNA scaffold t a g a g c t a g a a a t a g c a a g t t a a a a t a a g g c t a g t c c g t t a t c a a c t t g a a a a a g t g g c a a t c t c g a t c t t t a t c g t t c a a t t t t a t t c c g a t c a g g c a a t a g t t g a a c t t t t t c a c c g t

120 gRNA scaffold c c g a g t c g g t g c t t t t t t g t t t t a g a g c t a g a a a t a g c a a g t t a a a a t a a g g c t a g t c c g g g c t c a g c c a c g a a a a a a c a a a a t c t c g a t c t t t a t c g t t c a a t t t t a t t c c g a t c a g g c

XbaIAcc65IKpnI 180 CMV enhancer t t t t t a g c g c g t g c g c c a a t t c t g c a g a c a a a t g g c t c t a g a g g t a c c c g t t a c a t a a c t a a a a a t c g c g c a c g c g g t t a a g a c g t c t g t t t a c c g a g a t c t c c a t g g g c a a t g t a t t g a

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

300 CMV enhancer 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 a g t a t t t a c g g t a a a c t g c c 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 c t c a t a a a t g c c a t t t g a c g g g t

360 CMV enhancer 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 c c c c t a t t g a c g t c a a t g a c g g 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 g g g g g a t a a c t g c a g t t a c t g c c

420 CMV enhancer 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 t t a t g g g a c t t t c c t a c t t g g c a 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 a a t a c c c t g a a a g g a t g a a c c g t

SnaBI 480 CMV enhancer 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 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 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 c t c c a c t c g g g g t g c a a g a c g a a

540 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 t t g t a t t t a t t t a t t t t t t a a t t 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 a a c a t a a a t a a a t a a a a a a t t a a

600 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 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 g c a g c 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 g c c c c g c t c c g c c t c t c c a c g c c g c c g t c g

660 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 g c g g c c 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 g c c g c c g g

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

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

BseRI 840 hybrid intron 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 g a g g t a a g g 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 c t c c a t t c c

900 hybrid intron 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 c t g c c t g a 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 g g a c g g a c t

AgeIPaqCI 960 1 5 DYKDHDGD hybrid intron 3xFLAG Kozak sequence MDYKDHDGD *TPGTGGHVILVVVSV 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 a c g g a g a c 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 c t g c c t c t g

1020 10 15 20 YKDHDIDYKDDDDK MA PKKK 3xFLAG SV40 NLS YKDHDIDYKDDDDKMAPKKK VLIMINIVFVIVILHGWLLL 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 a a g a a g a a 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 t t c t t c t t c

1080 5 10 DKKYSIGLD Cas9 130 RKV (in frame with SV40 NLS) GIHGVPAA 5 10 DKKYSIGLD SV40 NLS Cas9 RKVGIHGVPAADKKYSIGLD PLDTDVSDWCGVLLVADAQV 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 g g c c t g g a 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 c c g g a c c t g

1140 15 20 25 30 IGTNSVGWAVITDEYKVPSK Cas9 15 20 25 30 IGTNSVGWAVITDEYKVPSK Cas9 IGTNSVGWAVITDEYKVPSK DAGVRHAPGHDGVLVLHGAL 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 c a g c a a 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 g g g t c g t t c

1200 35 40 45 50 KFKVLGNTDRHSIKKNLIGA Cas9 35 40 45 50 KFKVLGNTDRHSIKKNLIGA Cas9 KFKVLGNTDRHSIKKNLIGA FELHQAVGVPVADLLVQDSG 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 a t c g g a g c 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 t a g c c t c g g

1260 55 60 65 70 LLFDSGETAEATRLKRTARR Cas9 55 60 65 70 LLFDSGETAEATRLKRTARR Cas9 LLFDSGETAEATRLKRTARR QQEVAAFCGLGGPQLSGGSS 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 g c c a g a a g a 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 c g g t c t t c t

BglII 1320 75 80 85 90 RYTRRKNRICYLQEIFSNEM Cas9 75 80 85 90 RYTRRKNRICYLQEIFSNEM Cas9 RYTRRKNRICYLQEIFSNEM SVGSPLVPDAIQLLDEAVLH 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 a a c g a g a t g 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 t t g c t c t a c

1380 95 100 105 110 AKVDDSFFHRLEESFLVEED Cas9 95 100 105 110 AKVDDSFFHRLEESFLVEED Cas9 AKVDDSFFHRLEESFLVEED GLHVVAEEVSQFLGEQHFLI 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 g a a g a g g a t 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 c t t c t c c t a

1440 115 120 125 130 KKHERHPIFGNIVDEVAYHE Cas9 115 120 125 130 KKHERHPIFGNIVDEVAYHE Cas9 KKHERHPIFGNIVDEVAYHE LLVLPVGDEAVDHVLHGVVL 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 t a c c a c g a g 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 a t g g t g c t c

1500 135 140 145 150 KYPTIYHLRKKLVDSTDKAD Cas9 135 140 145 150 KYPTIYHLRKKLVDSTDKAD Cas9 KYPTIYHLRKKLVDSTDKAD LVGGDVVQSLFQHVAGVLGV 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 a g g c c g a 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 t t c c g g c t g

1560 155 160 165 170 LRLIYLALAHMIKFRGHFLI Cas9 155 160 165 170 LRLIYLALAHMIKFRGHFLI Cas9 LRLIYLALAHMIKFRGHFLI QPQDIQGQGVHDLEPAVEQD 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 t g a t c 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 a a g g a c t a g

1620 175 180 185 190 EGDLNPDNSDVDKLFIQLVQ Cas9 175 180 185 190 EGDLNPDNSDVDKLFIQLVQ Cas9 EGDLNPDNSDVDKLFIQLVQ LAVQVGVVAVHVLQEDLQHL 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 c t g g t g c 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 g a c c a c g t c

1680 195 200 205 210 TYNQLFEENPINASGVDAKA Cas9 195 200 205 210 TYNQLFEENPINASGVDAKA Cas9 TYNQLFEENPINASGVDAKA GVVLQELFVGDVGAAHVGLG 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 c c a a g g c 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 c g g t t c c g g

1740 215 220 225 230 ILSARLSKSRRLENLIAQLP Cas9 215 220 225 230 ILSARLSKSRRLENLIAQLP Cas9 ILSARLSKSRRLENLIAQLP DQRGSQALASPQFIQDGLQG 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 a g c t g c c c 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 g t c g a c g g g

1800 235 240 245 250 GEKKNGLFGNLIALSLGLTP Cas9 235 240 245 250 GEKKNGLFGNLIALSLGLTP Cas9 GEKKNGLFGNLIALSLGLTP ALLLIAQESVQNGQAQAQGG 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 c t g a c c c c c 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 g a c t g g g g g

1860 255 260 265 270 NFKSNFDLAEDAKLQLSKDT Cas9 255 260 265 270 NFKSNFDLAEDAKLQLSKDT Cas9 NFKSNFDLAEDAKLQLSKDT VELAVEVQGLIGFQLQALVG 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 a a g g a c a c c 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 t t c c t g t g g

1920 275 280 285 290 YDDDLDNLLAQIGDQYADLF Cas9 275 280 285 290 YDDDLDNLLAQIGDQYADLF Cas9 YDDDLDNLLAQIGDQYADLF VVVVQVVQQGLDAVLVGVQK 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 g a c c t g t t t 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 c t g g a c a a a

1980 295 300 305 310 LAAKNLSDAILLSDILRVNT Cas9 295 300 305 310 LAAKNLSDAILLSDILRVNT Cas9 LAAKNLSDAILLSDILRVNT QGGLVQGVGDQQAVDQSHVG 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 g t g a a c a c c 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 c a c t t g t g g

2040 315 320 325 330 EITKAPLSASMIKRYDEHHQ Cas9 315 320 325 330 EITKAPLSASMIKRYDEHHQ Cas9 EITKAPLSASMIKRYDEHHQ LDGLGGQAGRHDLSVVLVVL 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 c a c c a 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 g t g g t g g t c

2100 335 340 345 350 DLTLLKALVRQQLPEKYKEI Cas9 335 340 345 350 DLTLLKALVRQQLPEKYKEI Cas9 DLTLLKALVRQQLPEKYKEI VQGQQFSEHPLLQRLLVFLN 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 a a a g a g a t t 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 t t t c t c t a a

2160 355 360 365 370 FFDQSKNGYAGYIDGGASQE Cas9 355 360 365 370 FFDQSKNGYAGYIDGGASQE Cas9 FFDQSKNGYAGYIDGGASQE EEVLALVAVGAVNVASGALF 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 a g c c a g g 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 t c g g t c c t t

2220 375 380 385 390 EFYKFIKPILEKMDGTEELL Cas9 375 380 385 390 EFYKFIKPILEKMDGTEELL Cas9 EFYKFIKPILEKMDGTEELL LEVLEDLGDQFLHVAGLFQE 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 g a a c t g c t c 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 c t t g a c g a g

2280 395 400 405 410 VKLNREDLLRKQRTFDNGSI Cas9 395 400 405 410 VKLNREDLLRKQRTFDNGSI Cas9 VKLNREDLLRKQRTFDNGSI HLQVSLVQQPLLPGEVVAAD 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 g g c a g c a t c 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 c c g t c g t a g

2340 415 420 425 430 PHQIHLGELHAILRRQEDFY Cas9 415 420 425 430 PHQIHLGELHAILRRQEDFY Cas9 PHQIHLGELHAILRRQEDFY GVLDVQSLQVGNQPPLFIKV 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 g a t t t t t a c 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 c t a a a a a t g

2400 435 440 445 450 PFLKDNREKIEKILTFRIPY Cas9 435 440 445 450 PFLKDNREKIEKILTFRIPY Cas9 PFLKDNREKIEKILTFRIPY WEQLVVPFLDLLDQGEADGV 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 a t c c c c t a c 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 t a g g g g a t g

PspOMIApaI 2460 455 460 465 470 YVGPLARGNSRFAWMTRKSE Cas9 455 460 465 470 YVGPLARGNSRFAWMTRKSE Cas9 YVGPLARGNSRFAWMTRKSE VHARQGPSVASEGPHGSLAL 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 a a g a g c g a 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 t t c t c g c t c

2520 475 480 485 490 ETITPWNFEEVVDKGASAQS Cas9 475 480 485 490 ETITPWNFEEVVDKGASAQS Cas9 ETITPWNFEEVVDKGASAQS FGDGGPVELFHHVLASGGLA 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 g c c c a g a g 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 c g g g t c t c g

2580 495 500 505 510 FIERMTNFDKNLPNEKVLPK Cas9 495 500 505 510 FIERMTNFDKNLPNEKVLPK Cas9 FIERMTNFDKNLPNEKVLPK EDLPHGVEILVQGVLLHQGL t t c a t c g a g c g g a t g a c c a a 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 t g c c c a a g a a g t a g c t c g c c t a c t g g t t 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 g a c g g g t t c

2640 515 520 525 530 HSLLYEYFTVYNELTKVKYV Cas9 515 520 525 530 HSLLYEYFTVYNELTKVKYV Cas9 HSLLYEYFTVYNELTKVKYV VAQQVLVEGHIVLQGFHFVH 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 a a a t a c g t g 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 t t t a t g c a c

2700 535 540 545 550 TEGMRKPAFLSGEQKKAIVD Cas9 535 540 545 550 TEGMRKPAFLSGEQKKAIVD Cas9 TEGMRKPAFLSGEQKKAIVD GLSHSLGGEQAALLFLGDHV 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 a t c g t g g 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 t a g c a c c t g

2760 555 560 565 570 LLFKTNRKVTVKQLKEDYFK Cas9 555 560 565 570 LLFKTNRKVTVKQLKEDYFK Cas9 LLFKTNRKVTVKQLKEDYFK QQELGVPFHGHLLQFLVVEL 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 c t t c a a 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 a t g a a g t t c

2820 575 580 585 590 KIECFDSVEISGVEDRFNAS Cas9 575 580 585 590 KIECFDSVEISGVEDRFNAS Cas9 KIECFDSVEISGVEDRFNAS FDLAEVGHFDGAHFIPEVGG 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 a c g c c t c 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 t t g c g g a g g

PasI 2880 595 600 605 610 LGTYHDLLKIIKDKDFLDNE Cas9 595 600 605 610 LGTYHDLLKIIKDKDFLDNE Cas9 LGTYHDLLKIIKDKDFLDNE QACVVIQQFNDLVLVEQVIL 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 g a c a a t g a 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 c t g t t a c t c

EcoRV 2940 615 620 625 630 ENEDILEDIVLTLTLFEDRE Cas9 615 620 625 630 ENEDILEDIVLTLTLFEDRE Cas9 ENEDILEDIVLTLTLFEDRE FVLVNQFIDHQGQCQKLVSL 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 g a c a g a g a g 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 c t g t c t c t c

3000 635 640 645 650 MIEERLKTYAHLFDDKVMKQ Cas9 635 640 645 650 MIEERLKTYAHLFDDKVMKQ Cas9 MIEERLKTYAHLFDDKVMKQ *AWRNSSLTIFC HDLFPQFGIGVQEVVFHHLL 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 a t g a a g c a g 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 t a c t t c g t c

3060 655 660 665 670 LKRRRYTGWGRLSRKLINGI Cas9 655 660 665 670 LKRRRYTGWGRLSRKLINGI Cas9 LKRRRYTGWGRLSRKLINGI SFRRLYVPQPLSLRFSILPM QLPPSVGAPAPQAPLQDVAD 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 a g g c t g a g c c g g a a g c t g a t c a a c g g c a 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 t c c g a c t c g g c c t t c g a c t a g t t g c c g t a g

3120 675 680 685 690 RDKQSGKTILDFLKSDGFAN Cas9 675 680 685 690 RDKQSGKTILDFLKSDGFAN Cas9 RDKQSGKTILDFLKSDGFAN RSLCDPLVIRSKRFDSPKAL PVLLGALCDQIEQLGVAEGV c g g 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 c g c c a a c g c c 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 a a g c g g t t g

3180 695 700 705 710 RNFMQLIHDDSLTFKEDIQK Cas9 695 700 705 710 RNFMQLIHDDSLTFKEDIQK Cas9 RNFMQLIHDDSLTFKEDIQK LFKMCSIWSSLRVKLSSMWF SVEHLQDVVVAQGKFLVDLF a g a a a c t t c a t g c a 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 t c c a g a a a t c t t t g a a g t a c g t 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 t a g g t c t t t

3240 715 720 725 730 AQVSGQGDSLHEHIANLAGS Cas9 715 720 725 730 AQVSGQGDSLHEHIANLAGS Cas9 AQVSGQGDSLHEHIANLAGS AWTDPWPSLRCSCMALRAPL GLHGALAIAQVLVNGIQGAA 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 g g c c g g c a g c 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 c c g g c c g t c g

3300 735 740 745 750 PAIKKGILQTVKVVDELVKV Cas9 735 740 745 750 PAIKKGILQTVKVVDELVKV Cas9 PAIKKGILQTVKVVDELVKV GAMLFPMRCVTFTTSSSTFT GGNLLADQLCHLHHVLEHFH c c c g c c a t t a a g a a g g g c a t c c t g 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 c g t g a a a g t g g g g c g g t a a t t c t t c c c g t a g g a c 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 g c a c t t t c a c

3360 755 760 765 770 MGRHKPENIVIEMARENQTT Cas9 755 760 765 770 MGRHKPENIVIEMARENQTT Cas9 MGRHKPENIVIEMARENQTT IPRCLGSFMTISIALSFWVV HAPVLGLVDHDFHGSLVLGG 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 g a c c a c 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 g t c t g g t g g

3420 775 780 785 790 QKGQKNSRERMKRIEEGIKE Cas9 775 780 785 790 QKGQKNSRERMKRIEEGIKE Cas9 QKGQKNSRERMKRIEEGIKE WFPCFFLRSLIFRISSPM LLSLLVAALSHLPDFLADFL c a g a a g g g a c a g a a g a a c a g c 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 a t c a a a g a g g t c t t c c c t g t c t t c t t g t c g 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 t a g t t t c t c

3480 795 800 805 810 LGSQILKEHPVENTQLQNEK Cas9 795 800 805 810 LGSQILKEHPVENTQLQNEK Cas9 LGSQILKEHPVENTQLQNEK QAALDQFFVGHFVGLQLVLL 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 g c a g a a c g a g a a g 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 c g t c t t g c t c t t c

3540 815 820 825 830 LYLYYLQNGRDMYVDQELDI Cas9 815 820 825 830 LYLYYLQNGRDMYVDQELDI Cas9 LYLYYLQNGRDMYVDQELDI QVQVVQLIPPIHVHVLFQVD c t g t a c c t g t a c t a c c t g 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 c t g g a c a t c g a c a t g g a c a t g a t g g a c 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 g a c c t g t a g

3600 835 840 845 850 NRLSDYDVDHIVPQSFLKDD Cas9 835 840 845 850 NRLSDYDVDHIVPQSFLKDD Cas9 NRLSDYDVDHIVPQSFLKDD VPQGVVIHVM 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 g a a g g a c g a c 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 c t t c c t g c t g

3660 855 860 865 870 SIDNKVLTRSDKNRGKSDNV Cas9 855 860 865 870 SIDNKVLTRSDKNRGKSDNV Cas9 SIDNKVLTRSDKNRGKSDNV 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 g a c a a c g 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 c t g t t g c a c

3720 875 880 885 890 PSEEVVKKMKNYWRQLLNAK Cas9 875 880 885 890 PSEEVVKKMKNYWRQLLNAK Cas9 PSEEVVKKMKNYWRQLLNAK 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 a a c g c c a a g 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 t t g c g g t t c

3780 895 900 905 910 LITQRKFDNLTKAERGGLSE Cas9 895 900 905 910 LITQRKFDNLTKAERGGLSE Cas9 LITQRKFDNLTKAERGGLSE 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 c t g a g c g a a 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 g a c t c g c t t

PmlI 3840 915 920 925 930 LDKAGFIKRQLVETRQITKH Cas9 915 920 925 930 LDKAGFIKRQLVETRQITKH Cas9 LDKAGFIKRQLVETRQITKH 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 c a g a t c a c a a a g c a c 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 g t c t a g t g t t t c g t g

3900 935 940 945 950 VAQILDSRMNTKYDENDKLI Cas9 935 940 945 950 VAQILDSRMNTKYDENDKLI Cas9 VAQILDSRMNTKYDENDKLI 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 a a g c t g a t c 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 t t c g a c t a g

3960 955 960 965 970 REVKVITLKSKLVSDFRKDF Cas9 955 960 965 970 REVKVITLKSKLVSDFRKDF Cas9 REVKVITLKSKLVSDFRKDF 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 a a g g a t t t c 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 t t c c t a a a g

4020 975 980 985 990 QFYKVREINNYHHAHDAYLN Cas9 975 980 985 990 QFYKVREINNYHHAHDAYLN Cas9 QFYKVREINNYHHAHDAYLN 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 c t a c c t g a a c 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 g a t g g a c t t g

4080 995 1000 1005 1010 AVVGTALIKKYPKLESEFVY Cas9 995 1000 1005 1010 AVVGTALIKKYPKLESEFVY Cas9 AVVGTALIKKYPKLESEFVY 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 a a a g c g a g t t c g t g t a c 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 t t t c g c t c a a g c a c a t g

4140 1015 1020 1025 1030 GDYKVYDVRKMIAKSEQEIG Cas9 1015 1020 1025 1030 GDYKVYDVRKMIAKSEQEIG Cas9 GDYKVYDVRKMIAKSEQEIG g g c g a c t a c a a g g t g t a c g a c g t g 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 c c g c t g a t g t t c c a c a t g c t g c a c 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

4200 1035 1040 1045 1050 KATAKYFFYSNIMNFFKTEI Cas9 1035 1040 1045 1050 KATAKYFFYSNIMNFFKTEI Cas9 KATAKYFFYSNIMNFFKTEI a a g g c t a c c g c c 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 t t c c g a t g g c g g 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

4260 1055 1060 1065 1070 TLANGEIRKRPLIETNGETG Cas9 1055 1060 1065 1070 TLANGEIRKRPLIETNGETG Cas9 TLANGEIRKRPLIETNGETG a c c c t g g c c a a c 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 t g g g a c c g g t t 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

4320 1075 1080 1085 1090 EIVWDKGRDFATVRKVLSMP Cas9 1075 1080 1085 1090 EIVWDKGRDFATVRKVLSMP Cas9 EIVWDKGRDFATVRKVLSMP g a g a t c g t g t g g 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 t c t a g c a c a c 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

4380 1095 1100 1105 1110 QVNIVKKTEVQTGGFSKESI Cas9 1095 1100 1105 1110 QVNIVKKTEVQTGGFSKESI Cas9 QVNIVKKTEVQTGGFSKESI c a a g t g a a t a t c 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 g t t c a c t t a t a 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

KflI 4440 1115 1120 1125 1130 LPKRNSDKLIARKKDWDPKK Cas9 1115 1120 1125 1130 LPKRNSDKLIARKKDWDPKK Cas9 LPKRNSDKLIARKKDWDPKK c t g c c c a a g a g g 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 c c c t a a g a a g g a c g g g t t c t c 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 g g g a t t c t t c

4500 1135 1140 1145 1150 YGGFDSPTVAYSVLVVAKVE Cas9 1135 1140 1145 1150 YGGFDSPTVAYSVLVVAKVE Cas9 YGGFDSPTVAYSVLVVAKVE t a c g g c g g c t t c 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 t g c c g c c g a a g 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

BsaBI* 4560 1155 1160 1165 1170 KGKSKKLKSVKELLGITIME Cas9 1155 1160 1165 1170 KGKSKKLKSVKELLGITIME Cas9 KGKSKKLKSVKELLGITIME a a g g g c a a g t c c a a g a a a c t g 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 t t c c c g t t c a g g t t c t t t g a c 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

4620 1175 1180 1185 1190 RSSFEKNPIDFLEAKGYKEV Cas9 1175 1180 1185 1190 RSSFEKNPIDFLEAKGYKEV Cas9 RSSFEKNPIDFLEAKGYKEV a g a a g c a g c t t c 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 t c t t c g t c g a a 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

4680 1195 1200 1205 1210 KKDLIIKLPKYSLFELENGR Cas9 1195 1200 1205 1210 KKDLIIKLPKYSLFELENGR Cas9 KKDLIIKLPKYSLFELENGR a a a a a g g a c c t g a t c a t c a a g c t g c c t a a g t a c t c c c t g t t c g a g c t g g a a a a c g g c c g g t t t t t c c t g g a c t a g t a g t t c g a c g g a t t c a t g a g g g a c a a g c t c g a c c t t t t g c c g g c c

BsmI 4740 1215 1220 1225 1230 KRMLASAGELQKGNELALPS Cas9 1215 1220 1225 1230 KRMLASAGELQKGNELALPS Cas9 KRMLASAGELQKGNELALPS a a g a g a a t g c t g 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 t t c t c t t a c g 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

4800 1235 1240 1245 1250 KYVNFLYLASHYEKLKGSPE Cas9 1235 1240 1245 1250 KYVNFLYLASHYEKLKGSPE Cas9 KYVNFLYLASHYEKLKGSPE a a a t a t g t g a a c 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 t t t a t a c a c t t 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

4860 1255 1260 1265 1270 DNEQKQLFVEQHKHYLDEII Cas9 1255 1260 1265 1270 DNEQKQLFVEQHKHYLDEII Cas9 DNEQKQLFVEQHKHYLDEII g a t a a t g a g c a g 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 c t a t t a c t c g t 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

4920 1275 1280 1285 1290 EQISEFSKRVILADANLDKV Cas9 1275 1280 1285 1290 EQISEFSKRVILADANLDKV Cas9 EQISEFSKRVILADANLDKV g a g c a g a t c a g c 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 c g t c t a g t c g 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

4980 1295 1300 1305 1310 LSAYNKHRDKPIREQAENII Cas9 1295 1300 1305 1310 LSAYNKHRDKPIREQAENII Cas9 LSAYNKHRDKPIREQAENII c t g t c c g c c t a c 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 g a c a g g c g g a t g 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

5040 1315 1320 1325 1330 HLFTLTNLGAPAAFKYFDTT Cas9 1315 1320 1325 1330 HLFTLTNLGAPAAFKYFDTT Cas9 HLFTLTNLGAPAAFKYFDTT c a c c t g t t t a c c 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 g t g g a c a a a t g 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

5100 1335 1340 1345 1350 IDRKRYTSTKEVLDATLIHQ Cas9 1335 1340 1345 1350 IDRKRYTSTKEVLDATLIHQ Cas9 IDRKRYTSTKEVLDATLIHQ a t c g a c c g g a a g 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 t a g c t g g c c t t 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

5160 1355 1360 1365 SITGLYETRIDLSQLGGD 1 KR Cas9 nucleoplasmin NLS 1355 1360 1365 SITGLYETRIDLSQLGGD KR Cas9 SITGLYETRIDLSQLGGDKR a g c a t c a c c g g c 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 t c g t a g t g g c 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

EcoRIFseI 5220 (in frame with Cas9) PAATKKAGQAKKKK* 5 10 15 PAATKKAGQAKKKK * nucleoplasmin NLS PAATKKAGQAKKKK* c c g g c g g c c a c g 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 g c c g c c g g t g c 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

5280 bGH poly(A) signal G G A G C T G A C A C G G A A G A T BGH-rev g c t g a t c a g c c t 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 c g a c t a g t c g 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

5340 bGH poly(A) signal t g c c t t c c t t g a 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 a c g g a a g g a a c 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

5400 bGH poly(A) signal t t g c a t c g c a t t 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 a a c g t a g c g t a a 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

NotI 5460 bGH poly(A) signal AAV2 ITR AAV2 ITR g c a a g g g g g a g g 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 c g t t c c c c c t c c 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

5520 AAV2 ITR AAV2 ITR t a g t g a t g g a g t 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 a t c a c t a c c t c a 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

5580 AAV2 ITR AAV2 ITR c a a a g g t c g c c c 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 t t t c c a g c g 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

SbfIKasINarISfoIPluTI 5640 AAV2 ITR AAV2 ITR g c t g c c t g c a g g 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 c g a c g g a c g t c 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

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

5760 f1 ori G A G G A A A F1ori-R 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 c a g c g c c t t a g c g c c c g c t c c t t t 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 g t c g c g g a a t c g c g g g c g a g g a a a

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

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

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

6000 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 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 c c a a a c t g g a a c a a c a c t c a a c t c 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 g g t t t g a c c t t g t t g t g a g t t g a g

PsiI 6060 f1 ori 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 c c g a t t t c g g t c t a t t g g t t a a a 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 c g g c t a a a g c c a g a t a a c c a a t t t

6120 f1 ori 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 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 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 a t t g t t t t a t a a t t g c a a a t g t t a

6180 f1 ori A T G T T A G A C G A G A C T A C G G C pRS-marker 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 c g c a t a g t t a a g c c a g c c c c g a c 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 g c g t a t c a a t t c g g t c g g g g c t g t

6240 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 t c t g c t c c c g g c a t c c g c t t a c a g 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 a g a c g a g g g c c g t a g g c g a a t g t c

6300 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' 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 g a g g t t t t c a c c g t c a t c a c c g a a 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 c t c c a a a a g t g g c a g t a g t g g c t t

6360 C G G A G C A C T A T G C G G A T A A pBRforEco 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 t t t a t a g g t t a a t g t c a t g a t a a 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 a a a t a t c c a a t t a c a g t a c t a t t a

6420 AmpR promoter 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 g t g c g c g g a a c c c c t a t t t g 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 t t t a c a c g c g c c t t g g g g a t a a a c

6480 AmpR promoter 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 c a t g a g a c a a t a a c c c t g a t a a a t 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 g t a c t c t g t t a t t g g g a c t a t t t a

6540 1 5 10 signal sequence MSIQHFRVALI AmpR promoter AmpR MSIQHFRVALI 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 t c a a c a t t t c c g t g t c g c c c t t a t 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 a g t t g t a a a g g c a c a g c g g g a a t a

6600 15 20 signal sequence PFFAAFCLPVFA 25 30 HPETLVKV AmpR PFFAAFCLPVFAHPETLVKV 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 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 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 a g t g g g t c t t t g c g a c c a c t t t c a

6660 35 40 45 50 KDAEDQLGARVGYIELDLNS AmpR KDAEDQLGARVGYIELDLNS 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 t t a c a t c g a a c t g g a t c t c a a c a g 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 a a t g t a g c t t g a c c t a g a g t t g t c

6720 55 60 65 70 GKILESFRPEERFPMMSTFK AmpR GKILESFRPEERFPMMSTFK 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 t t t t c c a a t g a t g a g c a c t t t t a a 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 a a a a g g t t a c t a c t c g t g a a a a t t

6780 75 80 85 90 VLLCGAVLSRIDAGQEQLGR AmpR C G A T A C A C C G C G C C A T A A T A Amp-R VLLCGAVLSRIDAGQEQLGR 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 c g c c g g g c a a g a g c a a c t c g g t c g 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 g c g g c c c g t t c t c g t t g a g c c a g c

6840 95 100 105 110 RIHYSQNDLVEYSPVTEKHL AmpR RIHYSQNDLVEYSPVTEKHL 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 c t c a c c a g t c a c a g a a a a g c a t c t 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 g a g t g g t c a g t g t c t t t t c g t a g a

6900 115 120 125 130 TDGMTVRELCSAAITMSDNT AmpR TDGMTVRELCSAAITMSDNT 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 t g c c a t a a c c a t g a g t g a t a a c a c 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 a c g g t a t t g g t a c t c a c t a t t g t g

PvuI 6960 135 140 145 150 AANLLLTTIGGPKELTAFLH AmpR AANLLLTTIGGPKELTAFLH 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 g a a g g a g c t a a c c g c t t t t t t g c a 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 c t t c c t c g a t t g g c g a a a a a a c g t

7020 155 160 165 170 NMGDHVTRLDRWEPELNEAI AmpR NMGDHVTRLDRWEPELNEAI 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 g g a a c c g g a g c t g a a t g a a g c c a 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 c c t t g g c c t c g a c t t a c t t c g g t a

FspI 7080 175 180 185 190 PNDERDTTMPVAMATTLRKL AmpR PNDERDTTMPVAMATTLRKL 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 a a t g g c a a c a a c g t t g c g c a a a c t 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 t t a c c g t t g t t g c a a c g c g t t t g a

7140 195 200 205 210 LTGELLTLASRQQLIDWMEA AmpR LTGELLTLASRQQLIDWMEA 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 a c a a t t a a t a g a c t g g a t g g a g g c 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 t g t t a a t t a t c t g a c c t a c c t c c g

7200 215 220 225 230 DKVAGPLLRSALPAGWFIAD AmpR DKVAGPLLRSALPAGWFIAD 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 c c g g c t g g c t g g t t t a t t g c t g a 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 a g g c c g a c c g a c c a a a t a a c g a c t

SacII 7260 235 240 245 250 KSGAGERGSRGIIAALGPDG AmpR KSGAGERGSRGIIAALGPDG t a a a t c t g g a g c 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 a t t t a g a c c t 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

7320 255 260 265 270 KPSRIVVIYTTGSQATMDER AmpR KPSRIVVIYTTGSQATMDER 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 a g t c a g g c a a c t a t g g a t g a a c g 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 c t c a g t c c g t t g a t a c c t a c t t g c

7380 275 280 285 NRQIAEIGASLIKHW* AmpR NRQIAEIGASLIKHW* 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 t a a g c a t t g g t a a c t g t c a g a c c a 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 a t t c g t a a c c a t t g a c a g t c t g g t

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

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

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

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

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

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

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

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

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

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

8040 ori G G G A A A C G C C T G pBR322ori-F 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 g g a g c t t c c a g g g g g a a a c g c c t 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 c c t c g a a g g t c c c c c t t t g c g g a c

8100 ori G T A T C T T T pBR322ori-F 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 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 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 t g a a c t c g c a g c t a a a a a c a c t a c

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

AflIIIPciI 8220 U6 promoter G A G G G C C T A T T T C C C A T G A T T hU6-F 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 g a g g g c c t a t t t c c c a t g a t t c c t t c a t a t t 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 c t c c c g g a t a a a g g g t a c t a a g g a a g t a t a a

8280 U6 promoter t g c a t a t a c g a t a c a a g g c t g t t a g a g a g a t a a t t g g a a t t a a t t t g a c t g t a a a c a c a a a c g t a t a t g c t a t g t t c c g a c a a t c t c t c t a t t a a c c t t a a t t a a a c t g a c a t t t g t g t t

8340 U6 promoter a g a t a t t a g t a c a a a a t a c g t g a c g t a g a a a g t a a t a a t t t c t t g g g t a g t t t g c a g t t t t c t a t a a t c a t g t t t t a t g c a c t g c a t c t t t c a t t a t t a a a g a a c c c a t c a a a c g t c a a a

8400 U6 promoter G A C T A T C A T A T G C T T A C C G T LKO.1 5' t a a a a t t a t g t t t t a a a a t g g a c t a t c a t a t g c t t a c c g t a a c t t g a a a g t a t t t c g a t t a t t t t a a t a c a a a a t t t t a c c t g a t a g t a t a c g a a t g g c a t t g a a c t t t c a t a a a g c t a a

8460 U6 promoter gRNA scaffold t c t t g g c t t t a t a t a t c t t g t g g a a a g g a c g a a a c a c c g a a t g t t c c t c a t g c c t c a a t g a g a a c c g a a a t a t a t a g a a c a c c t t t c c t g c t t t g t g g c t t a c a a g g a g t a c g g a g t t a c

3ʹ 5ʹ 8463 gRNA scaffold t t t a a a

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.