Addgene pZKSCAN4.1.0-gDNA - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pZKSCAN4.1.0-gDNA


Map View

LKO.1 5' (8407 .. 8426) hU6-F (8236 .. 8256) AflIII - PciI (8230) pBR322ori-F (8075 .. 8094) SacII (7277) FspI (7119) PvuI (6973) Amp-R (6773 .. 6792) pBRforEco (6364 .. 6382) pGEX 3' (6304 .. 6326) pRS-marker (6185 .. 6204) PsiI (6071) F1ori-F (6010 .. 6031) F1ori-R (5800 .. 5819) PluTI (5643) SfoI (5641) NarI (5640) KasI (5639) SbfI (5636) NotI (5491) BGH-rev (5276 .. 5293) EcoRI (5252) FseI (5229) nucleoplasmin NLS BsmI (4737) BsaBI * (4595) XbaI (157) Acc65I (163) KpnI (167) SnaBI (430) chicken β-actin promoter BseRI (841) PaqCI (946) AgeI (968) SV40 NLS BglII (1347) PspOMI (2452) ApaI (2456) PasI (2866) EcoRV (2950) PmlI (3886) KflI (4474) pZKSCAN4.1.0-gDNA 8508 bp

Sequence View

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

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

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

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

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

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

PaqCI 960 hybrid intron * 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 a a t c a c t t t t t t t c 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 t t a g t g a a a a a a a g

AgeI 1020 1 5 10 DYKDHDGDYKDHD hybrid intron 3xFLAG Kozak sequence MDYKDHDGDYKDHD TPGTGGHVILVVVSVVLIMI 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 a c a a g g a t c a t g a 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 a t g t t c c t a g t a c t

1080 15 20 IDYKDDDDK MA 130 PKKKRKV GI 3xFLAG SV40 NLS IDYKDDDDKMAPKKKRKVGI NIVFVIVILHGWLLLPLDTD 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 c g g a a g g t c g g t a 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 g c c t t c c a g c c a t a

1140 5 10 15 DKKYSIGLDIGTNS Cas9 (in frame with SV40 NLS) HGVPAA 5 10 15 DKKYSIGLDIGTNS Cas9 HGVPAADKKYSIGLDIGTNS VSDWCGVLLVADAQVDAGVR 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 a t c g g c a c c a a c t c 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 t a g c c g t g g t t g a g

1200 20 25 30 35 VGWAVITDEYKVPSKKFKVL Cas9 20 25 30 35 VGWAVITDEYKVPSKKFKVL Cas9 VGWAVITDEYKVPSKKFKVL HAPGHDGVLVLHGALFELHQ 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 a a a t t c a a g g t g c t 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 t t t a a g t t c c a c g a

1260 40 45 50 55 GNTDRHSIKKNLIGALLFDS Cas9 40 45 50 55 GNTDRHSIKKNLIGALLFDS Cas9 GNTDRHSIKKNLIGALLFDS AVGVPVADLLVQDSGQQEVA 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 c t g c t g t t c g a c a g 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 g a c g a c a a g c t g t c

1320 60 65 70 75 GETAEATRLKRTARRRYTRR Cas9 60 65 70 75 GETAEATRLKRTARRRYTRR Cas9 GETAEATRLKRTARRRYTRR AFCGLGGPQLSGGSSSVGSP 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 a g a t a c a c c a g a c g 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 t c t a t g t g g t c t g c

BglII 1380 80 85 90 95 KNRICYLQEIFSNEMAKVDD Cas9 80 85 90 95 KNRICYLQEIFSNEMAKVDD Cas9 KNRICYLQEIFSNEMAKVDD LVPDAIQLLDEAVLHGLHVV 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 g c c a a g g t g g a c g a 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 c g g t t c c a c c t g c t

1440 100 105 110 115 SFFHRLEESFLVEEDKKHER Cas9 100 105 110 115 SFFHRLEESFLVEEDKKHER Cas9 SFFHRLEESFLVEEDKKHER AEEVSQFLGEQHFLILLVLP 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 a a g a a g c a c g a g c g 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 t t c t t c g t g c t c g c

1500 120 125 130 135 HPIFGNIVDEVAYHEKYPTI Cas9 120 125 130 135 HPIFGNIVDEVAYHEKYPTI Cas9 HPIFGNIVDEVAYHEKYPTI VGDEAVDHVLHGVVLLVGGD 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 a a g t a c c c c a c c a t 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 t t c a t g g g g t g g t a

1560 140 145 150 155 YHLRKKLVDSTDKADLRLIY Cas9 140 145 150 155 YHLRKKLVDSTDKADLRLIY Cas9 YHLRKKLVDSTDKADLRLIY VVQSLFQHVAGVLGVQPQDI 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 c t g c g g c t g a t c 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 g a c g c c g a c t a g a t

1620 160 165 170 175 LALAHMIKFRGHFLIEGDLN Cas9 160 165 170 175 LALAHMIKFRGHFLIEGDLN Cas9 LALAHMIKFRGHFLIEGDLN QGQGVHDLEPAVEQDLAVQV 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 g g g c g a c c t g a a 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 c t c c c g c t g g a c t t

1680 180 185 190 195 PDNSDVDKLFIQLVQTYNQL Cas9 180 185 190 195 PDNSDVDKLFIQLVQTYNQL Cas9 PDNSDVDKLFIQLVQTYNQL GVVAVHVLQEDLQHLGVVLQ 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 a c c t a c a a c c a g c 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 t g g a t g t t g g t c g a

1740 200 205 210 215 FEENPINASGVDAKAILSAR Cas9 200 205 210 215 FEENPINASGVDAKAILSAR Cas9 FEENPINASGVDAKAILSAR ELFVGDVGAAHVGLGDQRGS 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 a t c c t g t c t g c c a g 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 t a g g a c a g a c g g t c

1800 220 225 230 235 LSKSRRLENLIAQLPGEKKN Cas9 220 225 230 235 LSKSRRLENLIAQLPGEKKN Cas9 LSKSRRLENLIAQLPGEKKN QALASPQFIQDGLQGALLLI 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 g g c g a g a a g a a g a a 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 c c g c t c t t c t t c t t

1860 240 245 250 255 GLFGNLIALSLGLTPNFKSN Cas9 240 245 250 255 GLFGNLIALSLGLTPNFKSN Cas9 GLFGNLIALSLGLTPNFKSN AQESVQNGQAQAQGGVELAV 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 a a c t t c a a g a g c a a 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 t t g a a g t t c t c g t t

1920 260 265 270 275 FDLAEDAKLQLSKDTYDDDL Cas9 260 265 270 275 FDLAEDAKLQLSKDTYDDDL Cas9 FDLAEDAKLQLSKDTYDDDL EVQGLIGFQLQALVGVVVVQ 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 a c g a c g a c g a c c 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 a t g c t g c t g c t g g a

1980 280 285 290 295 DNLLAQIGDQYADLFLAAKN Cas9 280 285 290 295 DNLLAQIGDQYADLFLAAKN Cas9 DNLLAQIGDQYADLFLAAKN VVQQGLDAVLVGVQKQGGLV 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 c t g g c c g c c a a g a 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 g a c c g g c g g t t c t t

2040 300 305 310 315 LSDAILLSDILRVNTEITKA Cas9 300 305 310 315 LSDAILLSDILRVNTEITKA Cas9 LSDAILLSDILRVNTEITKA QGVGDQQAVDQSHVGLDGLG 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 g a t c a c c a a g g c 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 c t c t a g t g g t t c c g

2100 320 325 330 335 PLSASMIKRYDEHHQDLTLL Cas9 320 325 330 335 PLSASMIKRYDEHHQDLTLL Cas9 PLSASMIKRYDEHHQDLTLL GQAGRHDLSVVLVVLVQGQQ 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 g a c c t g a c c c t g c t 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 c t g g a c t g g g a c g a

2160 340 345 350 355 KALVRQQLPEKYKEIFFDQS Cas9 340 345 350 355 KALVRQQLPEKYKEIFFDQS Cas9 KALVRQQLPEKYKEIFFDQS FSEHPLLQRLLVFLNEEVLA 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 t t c t t c g a c c a g a g 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 a a g a a g c t g g t c t c

2220 360 365 370 375 KNGYAGYIDGGASQEEFYKF Cas9 360 365 370 375 KNGYAGYIDGGASQEEFYKF Cas9 KNGYAGYIDGGASQEEFYKF LVAVGAVNVASGALFLEVLE 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 g a g t t c t a c a a g t t 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 c t c a a g a t g t t c a a

2280 380 385 390 395 IKPILEKMDGTEELLVKLNR Cas9 380 385 390 395 IKPILEKMDGTEELLVKLNR Cas9 IKPILEKMDGTEELLVKLNR DLGDQFLHVAGLFQEHLQVS 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 g t g a a g c t g a a c a g 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 c a c t t c g a c t t g t c

2340 400 405 410 415 EDLLRKQRTFDNGSIPHQIH Cas9 400 405 410 415 EDLLRKQRTFDNGSIPHQIH Cas9 EDLLRKQRTFDNGSIPHQIH LVQQPLLPGEVVAADGVLDV 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 c c c a c c a g a t c c a 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 g g g g t g g t c t a g g t

2400 420 425 430 435 LGELHAILRRQEDFYPFLKD Cas9 420 425 430 435 LGELHAILRRQEDFYPFLKD Cas9 LGELHAILRRQEDFYPFLKD QSLQVGNQPPLFIKVWEQLV 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 c c a t t c c t g a a g g a 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 g g t a a g g a c t t c c t

PspOMIApaI 2460 440 445 450 455 NREKIEKILTFRIPYYVGPL Cas9 440 445 450 455 NREKIEKILTFRIPYYVGPL Cas9 NREKIEKILTFRIPYYVGPL VPFLDLLDQGEADGVVHARQ 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 t a c g t g g g c c c t 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 a t g c a c c c g g g a g a

2520 460 465 470 475 ARGNSRFAWMTRKSEETITP Cas9 460 465 470 475 ARGNSRFAWMTRKSEETITP Cas9 ARGNSRFAWMTRKSEETITP GPSVASEGPHGSLALFGDGG 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 g a a a c c a t c a c c c c 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 c t t t g g t a g t g g g g

2580 480 485 490 495 WNFEEVVDKGASAQSFIERM Cas9 480 485 490 495 WNFEEVVDKGASAQSFIERM Cas9 WNFEEVVDKGASAQSFIERM PVELFHHVLASGGLAEDLPH 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 t t c a t c g a g c g g a t 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 a a g t a g c t c g c c t a

2640 500 505 510 515 TNFDKNLPNEKVLPKHSLLY Cas9 500 505 510 515 TNFDKNLPNEKVLPKHSLLY Cas9 TNFDKNLPNEKVLPKHSLLY GVEILVQGVLLHQGLVAQQV 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 c a c a g c c t g c t g 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 g t g t c g g a c g a c a t

2700 520 525 530 535 EYFTVYNELTKVKYVTEGMR Cas9 520 525 530 535 EYFTVYNELTKVKYVTEGMR Cas9 EYFTVYNELTKVKYVTEGMR LVEGHIVLQGFHFVHGLSHS 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 a c c g a g g g a a t g a g 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 t g g c t c c c t t a c t c

2760 540 545 550 555 KPAFLSGEQKKAIVDLLFKT Cas9 540 545 550 555 KPAFLSGEQKKAIVDLLFKT Cas9 KPAFLSGEQKKAIVDLLFKT LGGEQAALLFLGDHVQQELG 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 c t g c t g t t c a a g a 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 g a c g a c a a g t t c t g

2820 560 565 570 575 NRKVTVKQLKEDYFKKIECF Cas9 560 565 570 575 NRKVTVKQLKEDYFKKIECF Cas9 NRKVTVKQLKEDYFKKIECF VPFHGHLLQFLVVELFDLAE 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 a a a a t c g a g t g c t t 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 t t t t a g c t c a c g a a

PasI 2880 580 585 590 595 DSVEISGVEDRFNASLGTYH Cas9 580 585 590 595 DSVEISGVEDRFNASLGTYH Cas9 DSVEISGVEDRFNASLGTYH VGHFDGAHFIPEVGGQACVV 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 c t g g g c a c a t a c c 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 g a c c c g t g t a t g g t

2940 600 605 610 615 DLLKIIKDKDFLDNEENEDI Cas9 600 605 610 615 DLLKIIKDKDFLDNEENEDI Cas9 DLLKIIKDKDFLDNEENEDI IQQFNDLVLVEQVILFVLVN 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 a a a c g a g g a c a 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 c t t t t g c t c c t g t a

EcoRV 3000 620 625 630 635 LEDIVLTLTLFEDREMIEER Cas9 620 625 630 635 LEDIVLTLTLFEDREMIEER Cas9 LEDIVLTLTLFEDREMIEER QFIDHQGQCQKLVSLHDLFP 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 a t g a t c g a g g a a c g 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 t a c t a g c t c c t t g c

3060 640 645 650 655 LKTYAHLFDDKVMKQLKRRR Cas9 640 645 650 655 LKTYAHLFDDKVMKQLKRRR Cas9 LKTYAHLFDDKVMKQLKRRR QFGIGVQEVVFHHLLQLPPS *AWRNSSLTIFCSFRRL 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 c t g a a g c g g c g g a g 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 g a c t t c g c c g c c t c

3120 660 665 670 675 YTGWGRLSRKLINGIRDKQS Cas9 660 665 670 675 YTGWGRLSRKLINGIRDKQS Cas9 YTGWGRLSRKLINGIRDKQS VGAPAPQAPLQDVADPVLLG YVPQPLSLRFSILPMRSLCD 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 c g g g a c a a g c a g 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 g c c c t g t t c g t c a g

3180 680 685 690 695 GKTILDFLKSDGFANRNFMQ Cas9 680 685 690 695 GKTILDFLKSDGFANRNFMQ Cas9 GKTILDFLKSDGFANRNFMQ ALCDQIEQLGVAEGVSVEHL PLVIRSKRFDSPKALLFKMC 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 a g a a a c t t c a t g c a 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 t c t t t g a a g t a c g t

3240 700 705 710 715 LIHDDSLTFKEDIQKAQVSG Cas9 700 705 710 715 LIHDDSLTFKEDIQKAQVSG Cas9 LIHDDSLTFKEDIQKAQVSG QDVVVAQGKFLVDLFGLHGA SIWSSLRVKLSSMWFAWTDP 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 g c c c a g g t g t c c g 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 t a g g t c t t t c g g g t c c a c a g g c c

3300 720 725 730 735 QGDSLHEHIANLAGSPAIKK Cas9 720 725 730 735 QGDSLHEHIANLAGSPAIKK Cas9 QGDSLHEHIANLAGSPAIKK LAIAQVLVNGIQGAAGGNLL WPSLRCSCMALRAPLGAMLF 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 c c g c c a t t a a g a a 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 g g g c g g t a a t t c t t

3360 740 745 750 755 GILQTVKVVDELVKVMGRHK Cas9 740 745 750 755 GILQTVKVVDELVKVMGRHK Cas9 GILQTVKVVDELVKVMGRHK ADQLCHLHHVLEHFHHAPVL PMRCVTFTTSSSTFTIPRCL 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 a t g g g c c g g c a c a a 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 t a c c c g g c c g t g t t

3420 760 765 770 775 PENIVIEMARENQTTQKGQK Cas9 760 765 770 775 PENIVIEMARENQTTQKGQK Cas9 PENIVIEMARENQTTQKGQK GLVDHDFHGSLVLGGLLSLL GSFMTISIALSFWVVWFPCF 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 c a g a a g g g a c a g a a 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 g t c t t c c c t g t c t t

3480 780 785 790 795 NSRERMKRIEEGIKELGSQI Cas9 780 785 790 795 NSRERMKRIEEGIKELGSQI Cas9 NSRERMKRIEEGIKELGSQI VAALSHLPDFLADFLQAALD FLRSLIFRISSPM 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 c t g g g c a g c c a g a 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 g a c c c g t c g g t c t a

3540 800 805 810 815 LKEHPVENTQLQNEKLYLYY Cas9 800 805 810 815 LKEHPVENTQLQNEKLYLYY Cas9 LKEHPVENTQLQNEKLYLYY QFFVGHFVGLQLVLLQVQVV 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 c t g t a c c t g t a 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 g a c a t g g a c a t g a t

3600 820 825 830 835 LQNGRDMYVDQELDINRLSD Cas9 820 825 830 835 LQNGRDMYVDQELDINRLSD Cas9 LQNGRDMYVDQELDINRLSD QLIPPIHVHVLFQVDVPQGV 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 a a c c g g c t g t c c g a 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 t t g g c c g a c a g g c t

3660 840 845 850 855 YDVDHIVPQSFLKDDSIDNK Cas9 840 845 850 855 YDVDHIVPQSFLKDDSIDNK Cas9 YDVDHIVPQSFLKDDSIDNK VIHVM 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 c c a t c g a c a a c a a 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 a g g t a g c t g t t g t t

3720 860 865 870 875 VLTRSDKNRGKSDNVPSEEV Cas9 860 865 870 875 VLTRSDKNRGKSDNVPSEEV Cas9 VLTRSDKNRGKSDNVPSEEV 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 c c c t c c g a a g a g g 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 g g g a g g c t t c t c c a

3780 880 885 890 895 VKKMKNYWRQLLNAKLITQR Cas9 880 885 890 895 VKKMKNYWRQLLNAKLITQR Cas9 VKKMKNYWRQLLNAKLITQR 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 c t g a t t a c c c a g a g 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 g a c t a a t g g g t c t c

3840 900 905 910 915 KFDNLTKAERGGLSELDKAG Cas9 900 905 910 915 KFDNLTKAERGGLSELDKAG Cas9 KFDNLTKAERGGLSELDKAG 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 c t g g a t a a g g c c g g 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 g a c c t a t t c c g g c c

PmlI 3900 920 925 930 935 FIKRQLVETRQITKHVAQIL Cas9 920 925 930 935 FIKRQLVETRQITKHVAQIL Cas9 FIKRQLVETRQITKHVAQIL 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 t g g c a c a g a t c c t 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 c a c c g t g t c t a g g a

3960 940 945 950 955 DSRMNTKYDENDKLIREVKV Cas9 940 945 950 955 DSRMNTKYDENDKLIREVKV Cas9 DSRMNTKYDENDKLIREVKV 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 g g g a a g t g a a a g t 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 g c c c t t c a c t t t c a

4020 960 965 970 975 ITLKSKLVSDFRKDFQFYKV Cas9 960 965 970 975 ITLKSKLVSDFRKDFQFYKV Cas9 ITLKSKLVSDFRKDFQFYKV 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 c a g t t t t a c a a a g t 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 g t c a a a a t g t t t c a

4080 980 985 990 995 REINNYHHAHDAYLNAVVGT Cas9 980 985 990 995 REINNYHHAHDAYLNAVVGT Cas9 REINNYHHAHDAYLNAVVGT 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 c c g t c g t g g g a a c 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 c g g c a g c a c c c t t g

4140 1000 1005 1010 1015 ALIKKYPKLESEFVYGDYKV Cas9 1000 1005 1010 1015 ALIKKYPKLESEFVYGDYKV Cas9 ALIKKYPKLESEFVYGDYKV 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 g g c g a c t a c a a g g t 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 c c g c t g a t g t t c c a

4200 1020 1025 1030 1035 YDVRKMIAKSEQEIGKATAK Cas9 1020 1025 1030 1035 YDVRKMIAKSEQEIGKATAK Cas9 YDVRKMIAKSEQEIGKATAK 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 a a g g c t a c c g c c a 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 t t c c g a t g g c g g t t

4260 1040 1045 1050 1055 YFFYSNIMNFFKTEITLANG Cas9 1040 1045 1050 1055 YFFYSNIMNFFKTEITLANG Cas9 YFFYSNIMNFFKTEITLANG 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 g g 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 c c

4320 1060 1065 1070 1075 EIRKRPLIETNGETGEIVWD Cas9 1060 1065 1070 1075 EIRKRPLIETNGETGEIVWD Cas9 EIRKRPLIETNGETGEIVWD 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 g a 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 c t

4380 1080 1085 1090 1095 KGRDFATVRKVLSMPQVNIV Cas9 1080 1085 1090 1095 KGRDFATVRKVLSMPQVNIV Cas9 KGRDFATVRKVLSMPQVNIV 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 g 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 c a

4440 1100 1105 1110 1115 KKTEVQTGGFSKESILPKRN Cas9 1100 1105 1110 1115 KKTEVQTGGFSKESILPKRN Cas9 KKTEVQTGGFSKESILPKRN 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 a 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 t t

KflI 4500 1120 1125 1130 1135 SDKLIARKKDWDPKKYGGFD Cas9 1120 1125 1130 1135 SDKLIARKKDWDPKKYGGFD Cas9 SDKLIARKKDWDPKKYGGFD 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 t a c g g c g g c t t c g a 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 a t g c c g c c g a a g c t

4560 1140 1145 1150 1155 SPTVAYSVLVVAKVEKGKSK Cas9 1140 1145 1150 1155 SPTVAYSVLVVAKVEKGKSK Cas9 SPTVAYSVLVVAKVEKGKSK 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 a a 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 t t

BsaBI* 4620 1160 1165 1170 1175 KLKSVKELLGITIMERSSFE Cas9 1160 1165 1170 1175 KLKSVKELLGITIMERSSFE Cas9 KLKSVKELLGITIMERSSFE 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 a g a a g c a g c t t c g a 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 t c t t c g t c g a a g c t

4680 1180 1185 1190 1195 KNPIDFLEAKGYKEVKKDLI Cas9 1180 1185 1190 1195 KNPIDFLEAKGYKEVKKDLI Cas9 KNPIDFLEAKGYKEVKKDLI 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 a 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 t a

BsmI 4740 1200 1205 1210 1215 IKLPKYSLFELENGRKRMLA Cas9 1200 1205 1210 1215 IKLPKYSLFELENGRKRMLA Cas9 IKLPKYSLFELENGRKRMLA 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 a a g a g a a t g c t g g c 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 t t c t c t t a c g a c c g

4800 1220 1225 1230 1235 SAGELQKGNELALPSKYVNF Cas9 1220 1225 1230 1235 SAGELQKGNELALPSKYVNF Cas9 SAGELQKGNELALPSKYVNF 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 t t 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 a a

4860 1240 1245 1250 1255 LYLASHYEKLKGSPEDNEQK Cas9 1240 1245 1250 1255 LYLASHYEKLKGSPEDNEQK Cas9 LYLASHYEKLKGSPEDNEQK 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 t t

4920 1260 1265 1270 1275 QLFVEQHKHYLDEIIEQISE Cas9 1260 1265 1270 1275 QLFVEQHKHYLDEIIEQISE Cas9 QLFVEQHKHYLDEIIEQISE 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 g a 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 c t

4980 1280 1285 1290 1295 FSKRVILADANLDKVLSAYN Cas9 1280 1285 1290 1295 FSKRVILADANLDKVLSAYN Cas9 FSKRVILADANLDKVLSAYN 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 a a 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 t t

5040 1300 1305 1310 1315 KHRDKPIREQAENIIHLFTL Cas9 1300 1305 1310 1315 KHRDKPIREQAENIIHLFTL Cas9 KHRDKPIREQAENIIHLFTL 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 c 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 g a

5100 1320 1325 1330 1335 TNLGAPAAFKYFDTTIDRKR Cas9 1320 1325 1330 1335 TNLGAPAAFKYFDTTIDRKR Cas9 TNLGAPAAFKYFDTTIDRKR 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 a g 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 t c

5160 1340 1345 1350 1355 YTSTKEVLDATLIHQSITGL Cas9 1340 1345 1350 1355 YTSTKEVLDATLIHQSITGL Cas9 YTSTKEVLDATLIHQSITGL 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 c t 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 g a

5220 1360 1365 YETRIDLSQLGGD 1 5 KRPAATK Cas9 nucleoplasmin NLS 1360 1365 YETRIDLSQLGGD (in frame with Cas9) KRPAATK Cas9 YETRIDLSQLGGDKRPAATK 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 a 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 t t

EcoRIFseI 5280 (in frame with Cas9) KAGQAKKKK* 10 15 KAGQAKKKK * nucleoplasmin NLS G G A G C BGH-rev KAGQAKKKK* 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 c g 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 g c

5340 bGH poly(A) signal T G A C A C G G A A G A T BGH-rev 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 c 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 g g

5400 bGH poly(A) signal 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 t 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 c a

5460 bGH poly(A) signal 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 a t 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 t a

NotI 5520 bGH poly(A) signal AAV2 ITR AAV2 ITR 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 g 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 a c

5580 AAV2 ITR AAV2 ITR 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 g a 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 c t

SbfIKasI 5640 AAV2 ITR AAV2 ITR 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 g g 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 c c

NarISfoIPluTI 5700 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 a a 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 t t

5760 f1 ori 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 g c 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 c g

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

5880 f1 ori 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 g 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 t c

5940 f1 ori 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 t 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 a a

6000 f1 ori 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 g t 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 c a

6060 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 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 t t 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 a a

PsiI 6120 f1 ori 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 t t 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 a a

6180 f1 ori 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 t c 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 a g

6240 A T G T T A G A C G A G A C T A C G G C pRS-marker 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 c 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 g c

6300 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 c g 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 g c

6360 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' 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 a a 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 t t

6420 C G G A G C A C T A T G C G G A T A A pBRforEco 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 g a 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 c t

6480 AmpR promoter 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 a a 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 t t

6540 AmpR promoter 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 a a

6600 1 5 10 15 signal sequence MSIQHFRVALIPFFA AmpR promoter AmpR MSIQHFRVALIPFFA 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 g g 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 c c

6660 20 signal sequence AFCLPVFA 25 30 35 HPETLVKVKDAE AmpR AFCLPVFAHPETLVKVKDAE 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 a g 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 t c

6720 40 45 50 55 DQLGARVGYIELDLNSGKIL AmpR DQLGARVGYIELDLNSGKIL 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 g 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 a c

6780 60 65 70 75 ESFRPEERFPMMSTFKVLLC AmpR C G A T A C A C Amp-R ESFRPEERFPMMSTFKVLLC 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 t g 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 a c

6840 80 85 90 95 GAVLSRIDAGQEQLGRRIHY AmpR C G C G C C A T A A T A Amp-R GAVLSRIDAGQEQLGRRIHY 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 t t 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 a a

6900 100 105 110 115 SQNDLVEYSPVTEKHLTDGM AmpR SQNDLVEYSPVTEKHLTDGM 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 g 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 c t

6960 120 125 130 135 TVRELCSAAITMSDNTAANL AmpR TVRELCSAAITMSDNTAANL 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 a c 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 t g

PvuI 7020 140 145 150 155 LLTTIGGPKELTAFLHNMGD AmpR LLTTIGGPKELTAFLHNMGD 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 c 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 a g

7080 160 165 170 175 HVTRLDRWEPELNEAIPNDE AmpR HVTRLDRWEPELNEAIPNDE 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 g c 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 c g

FspI 7140 180 185 190 195 RDTTMPVAMATTLRKLLTGE AmpR RDTTMPVAMATTLRKLLTGE 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 a c 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 t g

7200 200 205 210 215 LLTLASRQQLIDWMEADKVA AmpR LLTLASRQQLIDWMEADKVA 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 a 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 t c

7260 220 225 230 235 GPLLRSALPAGWFIADKSGA AmpR GPLLRSALPAGWFIADKSGA 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 c g 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 g c

SacII 7320 240 245 250 255 GERGSRGIIAALGPDGKPSR AmpR GERGSRGIIAALGPDGKPSR 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 t a 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 a t

7380 260 265 270 275 IVVIYTTGSQATMDERNRQI AmpR IVVIYTTGSQATMDERNRQI 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 c g 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 g c

7440 280 285 AEIGASLIKHW* AmpR AEIGASLIKHW* 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 t a 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 a t

7500 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 t 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 a a

7560 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 c c 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 g g

7620 ori 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 c t 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 g a

7680 ori 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 a 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 t t

7740 ori 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 a g 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 t c

7800 ori 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 a g

7860 ori 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 g 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 c c

7920 ori 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 a 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 g t

7980 ori 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 a 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 t a

8040 ori 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 g g 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 c c

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

8160 ori 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 g c 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 c g

8220 ori 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 g c 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 c g

AflIIIPciI 8280 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 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 t g c a t a t a c g a t a c 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 a c g t a t a t g c t a t g

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

8400 U6 promoter 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 a a a a t t a t g t t 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 a t t t t a a t a c a a a a

8460 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' 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 t c t t g g c t t t a t a t 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 a g a a c c g a a a t a t a

8508 U6 promoter gRNA scaffold a t c t t g t g g a a a g g a c g a a a c a c c a a c t c t t t c a c a g t g a c c c a g t t t t a g a a c a c c t t t c c t g c t t t g t g g t t g a g a a a g t g t c a c t g g g t c 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.