Addgene pX330-EN1082_Rad21_STOP - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pX330-EN1082_Rad21_STOP


Map View

AflIII - PciI (8504) pBR322ori-F (8349 .. 8368) SacII (7551) FspI (7393) PvuI (7247) Amp-R (7047 .. 7066) pBRforEco (6638 .. 6656) pGEX 3' (6578 .. 6600) pRS-marker (6459 .. 6478) PsiI (6345) F1ori-F (6284 .. 6305) F1ori-R (6074 .. 6093) PluTI (5917) SfoI (5915) NarI (5914) KasI (5913) SbfI (5910) NotI (5765) BGH-rev (5550 .. 5567) EcoRI (5526) FseI (5503) nucleoplasmin NLS BsmI (5011) BsaBI * (4869) KflI (4748) hU6-F (1 .. 21) LKO.1 5' (172 .. 191) XbaI (431) Acc65I (437) KpnI (441) SnaBI (704) BseRI (1115) hybrid intron PaqCI (1220) AgeI (1242) 3xFLAG SV40 NLS BglII (1621) PspOMI (2726) ApaI (2730) PasI (3140) EcoRV (3224) PmlI (4160) pX330-EN1082_Rad21_STOP 8509 bp

Sequence View

60 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 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 a a g g c t g t t a g a g a g 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 t t c c g a c a a t c t c t c

120 U6 promoter 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 a a t a c g t g a c g t a g a 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 t t a t g c a c t g c a t c t

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

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

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

360 gRNA scaffold 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 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 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 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

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

XbaIAcc65IKpnI 480 CMV enhancer 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 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 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 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

540 CMV enhancer 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 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 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 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

600 CMV enhancer 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 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 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 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

660 CMV enhancer 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 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 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 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

SnaBI 720 CMV enhancer 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 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 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 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

780 CMV enhancer chicken β- actin promoter 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 c t c t c c c c a t c t c c c c c c c c t c c c 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 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

840 chicken β- actin promoter 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 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 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 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

900 chicken β- actin promoter 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 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 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 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

960 chicken β- actin promoter 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 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 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 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

1020 chicken β- actin promoter hybrid intron 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 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 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 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

1080 hybrid intron 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 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 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 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

BseRI 1140 hybrid intron 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 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 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 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

1200 hybrid intron 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 g t t g g t g g g g t a t t a a t g t t t a a t 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 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

AgeIPaqCI 1260 1 D hybrid intron 3xFLAG Kozak sequence MD *TPGTGGHV 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 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 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 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

1320 5 10 15 20 YKDHDGDYKDHDIDYKDDDD 3xFLAG YKDHDGDYKDHDIDYKDDDD ILVVVSVVLIMINIVFVIVI 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 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 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 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

1380 K MA 130 PKKKRKV (in frame with SV40 NLS) GIHGVPAA DK 3xFLAG SV40 NLS Cas9 DK Cas9 KMAPKKKRKVGIHGVPAADK LHGWLLLPLDTDVSDWCGVL 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 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 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 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

1440 5 10 15 20 KYSIGLDIGTNSVGWAVITD Cas9 5 10 15 20 KYSIGLDIGTNSVGWAVITD Cas9 KYSIGLDIGTNSVGWAVITD LVADAQVDAGVRHAPGHDGV 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 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 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 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

1500 25 30 35 40 EYKVPSKKFKVLGNTDRHSI Cas9 25 30 35 40 EYKVPSKKFKVLGNTDRHSI Cas9 EYKVPSKKFKVLGNTDRHSI LVLHGALFELHQAVGVPVAD 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 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 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 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

1560 45 50 55 60 KKNLIGALLFDSGETAEATR Cas9 45 50 55 60 KKNLIGALLFDSGETAEATR Cas9 KKNLIGALLFDSGETAEATR LLVQDSGQQEVAAFCGLGGP 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 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 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 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

1620 65 70 75 80 LKRTARRRYTRRKNRICYLQ Cas9 65 70 75 80 LKRTARRRYTRRKNRICYLQ Cas9 LKRTARRRYTRRKNRICYLQ QLSGGSSSVGSPLVPDAIQL 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 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 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 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

BglII 1680 85 90 95 100 EIFSNEMAKVDDSFFHRLEE Cas9 85 90 95 100 EIFSNEMAKVDDSFFHRLEE Cas9 EIFSNEMAKVDDSFFHRLEE LDEAVLHGLHVVAEEVSQFL 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 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 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 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

1740 105 110 115 120 SFLVEEDKKHERHPIFGNIV Cas9 105 110 115 120 SFLVEEDKKHERHPIFGNIV Cas9 SFLVEEDKKHERHPIFGNIV GEQHFLILLVLPVGDEAVDH 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 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 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 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

1800 125 130 135 140 DEVAYHEKYPTIYHLRKKLV Cas9 125 130 135 140 DEVAYHEKYPTIYHLRKKLV Cas9 DEVAYHEKYPTIYHLRKKLV VLHGVVLLVGGDVVQSLFQH 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 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 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 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

1860 145 150 155 160 DSTDKADLRLIYLALAHMIK Cas9 145 150 155 160 DSTDKADLRLIYLALAHMIK Cas9 DSTDKADLRLIYLALAHMIK VAGVLGVQPQDIQGQGVHDL 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 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 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 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

1920 165 170 175 180 FRGHFLIEGDLNPDNSDVDK Cas9 165 170 175 180 FRGHFLIEGDLNPDNSDVDK Cas9 FRGHFLIEGDLNPDNSDVDK EPAVEQDLAVQVGVVAVHVL 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 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 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 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

1980 185 190 195 200 LFIQLVQTYNQLFEENPINA Cas9 185 190 195 200 LFIQLVQTYNQLFEENPINA Cas9 LFIQLVQTYNQLFEENPINA QEDLQHLGVVLQELFVGDVG 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 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 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 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

2040 205 210 215 220 SGVDAKAILSARLSKSRRLE Cas9 205 210 215 220 SGVDAKAILSARLSKSRRLE Cas9 SGVDAKAILSARLSKSRRLE AAHVGLGDQRGSQALASPQF 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 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 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 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

2100 225 230 235 240 NLIAQLPGEKKNGLFGNLIA Cas9 225 230 235 240 NLIAQLPGEKKNGLFGNLIA Cas9 NLIAQLPGEKKNGLFGNLIA IQDGLQGALLLIAQESVQNG 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 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 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 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

2160 245 250 255 260 LSLGLTPNFKSNFDLAEDAK Cas9 245 250 255 260 LSLGLTPNFKSNFDLAEDAK Cas9 LSLGLTPNFKSNFDLAEDAK QAQAQGGVELAVEVQGLIGF 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 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 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 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

2220 265 270 275 280 LQLSKDTYDDDLDNLLAQIG Cas9 265 270 275 280 LQLSKDTYDDDLDNLLAQIG Cas9 LQLSKDTYDDDLDNLLAQIG QLQALVGVVVVQVVQQGLDA 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 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 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 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

2280 285 290 295 300 DQYADLFLAAKNLSDAILLS Cas9 285 290 295 300 DQYADLFLAAKNLSDAILLS Cas9 DQYADLFLAAKNLSDAILLS VLVGVQKQGGLVQGVGDQQA 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 c c g a c g c c a t c c t g c t g a g c g 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 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

2340 305 310 315 320 DILRVNTEITKAPLSASMIK Cas9 305 310 315 320 DILRVNTEITKAPLSASMIK Cas9 DILRVNTEITKAPLSASMIK VDQSHVGLDGLGGQAGRHDL 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 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 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 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

2400 325 330 335 340 RYDEHHQDLTLLKALVRQQL Cas9 325 330 335 340 RYDEHHQDLTLLKALVRQQL Cas9 RYDEHHQDLTLLKALVRQQL SVVLVVLVQGQQFSEHPLLQ 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 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 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 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

2460 345 350 355 360 PEKYKEIFFDQSKNGYAGYI Cas9 345 350 355 360 PEKYKEIFFDQSKNGYAGYI Cas9 PEKYKEIFFDQSKNGYAGYI RLLVFLNEEVLALVAVGAVN 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 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 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 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

2520 365 370 375 380 DGGASQEEFYKFIKPILEKM Cas9 365 370 375 380 DGGASQEEFYKFIKPILEKM Cas9 DGGASQEEFYKFIKPILEKM VASGALFLEVLEDLGDQFLH 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 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 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 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

2580 385 390 395 400 DGTEELLVKLNREDLLRKQR Cas9 385 390 395 400 DGTEELLVKLNREDLLRKQR Cas9 DGTEELLVKLNREDLLRKQR VAGLFQEHLQVSLVQQPLLP 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 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 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 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

2640 405 410 415 420 TFDNGSIPHQIHLGELHAIL Cas9 405 410 415 420 TFDNGSIPHQIHLGELHAIL Cas9 TFDNGSIPHQIHLGELHAIL GEVVAADGVLDVQSLQVGNQ 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 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 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 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

2700 425 430 435 440 RRQEDFYPFLKDNREKIEKI Cas9 425 430 435 440 RRQEDFYPFLKDNREKIEKI Cas9 RRQEDFYPFLKDNREKIEKI PPLFIKVWEQLVVPFLDLLD 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 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 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 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

PspOMIApaI 2760 445 450 455 460 LTFRIPYYVGPLARGNSRFA Cas9 445 450 455 460 LTFRIPYYVGPLARGNSRFA Cas9 LTFRIPYYVGPLARGNSRFA QGEADGVVHARQGPSVASEG 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 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 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 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

2820 465 470 475 480 WMTRKSEETITPWNFEEVVD Cas9 465 470 475 480 WMTRKSEETITPWNFEEVVD Cas9 WMTRKSEETITPWNFEEVVD PHGSLALFGDGGPVELFHHV 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 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 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 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

2880 485 490 495 500 KGASAQSFIERMTNFDKNLP Cas9 485 490 495 500 KGASAQSFIERMTNFDKNLP Cas9 KGASAQSFIERMTNFDKNLP LASGGLAEDLPHGVEILVQG 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 a a c t t c g a t a a g a a c c t g c c c a 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 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

2940 505 510 515 520 NEKVLPKHSLLYEYFTVYNE Cas9 505 510 515 520 NEKVLPKHSLLYEYFTVYNE Cas9 NEKVLPKHSLLYEYFTVYNE VLLHQGLVAQQVLVEGHIVL 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 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 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 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

3000 525 530 535 540 LTKVKYVTEGMRKPAFLSGE Cas9 525 530 535 540 LTKVKYVTEGMRKPAFLSGE Cas9 LTKVKYVTEGMRKPAFLSGE QGFHFVHGLSHSLGGEQAAL 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 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 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 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

3060 545 550 555 560 QKKAIVDLLFKTNRKVTVKQ Cas9 545 550 555 560 QKKAIVDLLFKTNRKVTVKQ Cas9 QKKAIVDLLFKTNRKVTVKQ LFLGDHVQQELGVPFHGHLL 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 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 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 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

3120 565 570 575 580 LKEDYFKKIECFDSVEISGV Cas9 565 570 575 580 LKEDYFKKIECFDSVEISGV Cas9 LKEDYFKKIECFDSVEISGV QFLVVELFDLAEVGHFDGAH 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 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 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 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

PasI 3180 585 590 595 600 EDRFNASLGTYHDLLKIIKD Cas9 585 590 595 600 EDRFNASLGTYHDLLKIIKD Cas9 EDRFNASLGTYHDLLKIIKD FIPEVGGQACVVIQQFNDLV 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 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 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 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

EcoRV 3240 605 610 615 620 KDFLDNEENEDILEDIVLTL Cas9 605 610 615 620 KDFLDNEENEDILEDIVLTL Cas9 KDFLDNEENEDILEDIVLTL LVEQVILFVLVNQFIDHQGQ 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 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 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 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

3300 625 630 635 640 TLFEDREMIEERLKTYAHLF Cas9 625 630 635 640 TLFEDREMIEERLKTYAHLF Cas9 TLFEDREMIEERLKTYAHLF CQKLVSLHDLFPQFGIGVQE *AWRNS 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 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 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 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

3360 645 650 655 660 DDKVMKQLKRRRYTGWGRLS Cas9 645 650 655 660 DDKVMKQLKRRRYTGWGRLS Cas9 DDKVMKQLKRRRYTGWGRLS VVFHHLLQLPPSVGAPAPQA SLTIFCSFRRLYVPQPLSLR 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 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 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 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

3420 665 670 675 680 RKLINGIRDKQSGKTILDFL Cas9 665 670 675 680 RKLINGIRDKQSGKTILDFL Cas9 RKLINGIRDKQSGKTILDFL PLQDVADPVLLGALCDQIEQ FSILPMRSLCDPLVIRSKRF 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 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 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 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

3480 685 690 695 700 KSDGFANRNFMQLIHDDSLT Cas9 685 690 695 700 KSDGFANRNFMQLIHDDSLT Cas9 KSDGFANRNFMQLIHDDSLT LGVAEGVSVEHLQDVVVAQG DSPKALLFKMCSIWSSLRVK 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 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 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 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

3540 705 710 715 720 FKEDIQKAQVSGQGDSLHEH Cas9 705 710 715 720 FKEDIQKAQVSGQGDSLHEH Cas9 FKEDIQKAQVSGQGDSLHEH KFLVDLFGLHGALAIAQVLV LSSMWFAWTDPWPSLRCSCM 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 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 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 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

3600 725 730 735 740 IANLAGSPAIKKGILQTVKV Cas9 725 730 735 740 IANLAGSPAIKKGILQTVKV Cas9 IANLAGSPAIKKGILQTVKV NGIQGAAGGNLLADQLCHLH ALRAPLGAMLFPMRCVTFTT 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 g c a t c c t g c a g a c a g t g a a g g t g g 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 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

3660 745 750 755 760 VDELVKVMGRHKPENIVIEM Cas9 745 750 755 760 VDELVKVMGRHKPENIVIEM Cas9 VDELVKVMGRHKPENIVIEM HVLEHFHHAPVLGLVDHDFH SSSTFTIPRCLGSFMTISIA 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 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 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 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

3720 765 770 775 780 ARENQTTQKGQKNSRERMKR Cas9 765 770 775 780 ARENQTTQKGQKNSRERMKR Cas9 ARENQTTQKGQKNSRERMKR GSLVLGGLLSLLVAALSHLP LSFWVVWFPCFFLRSLIFRI 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 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 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 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

3780 785 790 795 800 IEEGIKELGSQILKEHPVEN Cas9 785 790 795 800 IEEGIKELGSQILKEHPVEN Cas9 IEEGIKELGSQILKEHPVEN DFLADFLQAALDQFFVGHFV SSPM 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 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 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 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

3840 805 810 815 820 TQLQNEKLYLYYLQNGRDMY Cas9 805 810 815 820 TQLQNEKLYLYYLQNGRDMY Cas9 TQLQNEKLYLYYLQNGRDMY GLQLVLLQVQVVQLIPPIHV 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 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 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 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

3900 825 830 835 840 VDQELDINRLSDYDVDHIVP Cas9 825 830 835 840 VDQELDINRLSDYDVDHIVP Cas9 VDQELDINRLSDYDVDHIVP HVLFQVDVPQGVVIHVM 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 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 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 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

3960 845 850 855 860 QSFLKDDSIDNKVLTRSDKN Cas9 845 850 855 860 QSFLKDDSIDNKVLTRSDKN Cas9 QSFLKDDSIDNKVLTRSDKN 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 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 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 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

4020 865 870 875 880 RGKSDNVPSEEVVKKMKNYW Cas9 865 870 875 880 RGKSDNVPSEEVVKKMKNYW Cas9 RGKSDNVPSEEVVKKMKNYW 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 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 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 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

4080 885 890 895 900 RQLLNAKLITQRKFDNLTKA Cas9 885 890 895 900 RQLLNAKLITQRKFDNLTKA Cas9 RQLLNAKLITQRKFDNLTKA 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 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 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 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

4140 905 910 915 920 ERGGLSELDKAGFIKRQLVE Cas9 905 910 915 920 ERGGLSELDKAGFIKRQLVE Cas9 ERGGLSELDKAGFIKRQLVE 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 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 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 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

PmlI 4200 925 930 935 940 TRQITKHVAQILDSRMNTKY Cas9 925 930 935 940 TRQITKHVAQILDSRMNTKY Cas9 TRQITKHVAQILDSRMNTKY 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 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 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 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

4260 945 950 955 960 DENDKLIREVKVITLKSKLV Cas9 945 950 955 960 DENDKLIREVKVITLKSKLV Cas9 DENDKLIREVKVITLKSKLV 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 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 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 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

4320 965 970 975 980 SDFRKDFQFYKVREINNYHH Cas9 965 970 975 980 SDFRKDFQFYKVREINNYHH Cas9 SDFRKDFQFYKVREINNYHH 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 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 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 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

4380 985 990 995 1000 AHDAYLNAVVGTALIKKYPK Cas9 985 990 995 1000 AHDAYLNAVVGTALIKKYPK Cas9 AHDAYLNAVVGTALIKKYPK 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 c c t g a t c a a a a a g t a c c c t a a 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 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

4440 1005 1010 1015 1020 LESEFVYGDYKVYDVRKMIA Cas9 1005 1010 1015 1020 LESEFVYGDYKVYDVRKMIA Cas9 LESEFVYGDYKVYDVRKMIA 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 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 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 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

4500 1025 1030 1035 1040 KSEQEIGKATAKYFFYSNIM Cas9 1025 1030 1035 1040 KSEQEIGKATAKYFFYSNIM Cas9 KSEQEIGKATAKYFFYSNIM 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 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 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 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

4560 1045 1050 1055 1060 NFFKTEITLANGEIRKRPLI Cas9 1045 1050 1055 1060 NFFKTEITLANGEIRKRPLI Cas9 NFFKTEITLANGEIRKRPLI 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 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 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 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

4620 1065 1070 1075 1080 ETNGETGEIVWDKGRDFATV Cas9 1065 1070 1075 1080 ETNGETGEIVWDKGRDFATV Cas9 ETNGETGEIVWDKGRDFATV 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 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 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 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

4680 1085 1090 1095 1100 RKVLSMPQVNIVKKTEVQTG Cas9 1085 1090 1095 1100 RKVLSMPQVNIVKKTEVQTG Cas9 RKVLSMPQVNIVKKTEVQTG 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 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 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 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

4740 1105 1110 1115 1120 GFSKESILPKRNSDKLIARK Cas9 1105 1110 1115 1120 GFSKESILPKRNSDKLIARK Cas9 GFSKESILPKRNSDKLIARK 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 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 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 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

KflI 4800 1125 1130 1135 1140 KDWDPKKYGGFDSPTVAYSV Cas9 1125 1130 1135 1140 KDWDPKKYGGFDSPTVAYSV Cas9 KDWDPKKYGGFDSPTVAYSV 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 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 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 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

4860 1145 1150 1155 1160 LVVAKVEKGKSKKLKSVKEL Cas9 1145 1150 1155 1160 LVVAKVEKGKSKKLKSVKEL Cas9 LVVAKVEKGKSKKLKSVKEL 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 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 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 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

BsaBI* 4920 1165 1170 1175 1180 LGITIMERSSFEKNPIDFLE Cas9 1165 1170 1175 1180 LGITIMERSSFEKNPIDFLE Cas9 LGITIMERSSFEKNPIDFLE 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 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 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 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

4980 1185 1190 1195 1200 AKGYKEVKKDLIIKLPKYSL Cas9 1185 1190 1195 1200 AKGYKEVKKDLIIKLPKYSL Cas9 AKGYKEVKKDLIIKLPKYSL 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 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 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 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

BsmI 5040 1205 1210 1215 1220 FELENGRKRMLASAGELQKG Cas9 1205 1210 1215 1220 FELENGRKRMLASAGELQKG Cas9 FELENGRKRMLASAGELQKG 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 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 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 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

5100 1225 1230 1235 1240 NELALPSKYVNFLYLASHYE Cas9 1225 1230 1235 1240 NELALPSKYVNFLYLASHYE Cas9 NELALPSKYVNFLYLASHYE 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 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 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 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

5160 1245 1250 1255 1260 KLKGSPEDNEQKQLFVEQHK Cas9 1245 1250 1255 1260 KLKGSPEDNEQKQLFVEQHK Cas9 KLKGSPEDNEQKQLFVEQHK 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 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 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 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

5220 1265 1270 1275 1280 HYLDEIIEQISEFSKRVILA Cas9 1265 1270 1275 1280 HYLDEIIEQISEFSKRVILA Cas9 HYLDEIIEQISEFSKRVILA 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 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 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 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

5280 1285 1290 1295 1300 DANLDKVLSAYNKHRDKPIR Cas9 1285 1290 1295 1300 DANLDKVLSAYNKHRDKPIR Cas9 DANLDKVLSAYNKHRDKPIR 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 c a c c g g g a t a a g c c c a t c a g a g 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 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

5340 1305 1310 1315 1320 EQAENIIHLFTLTNLGAPAA Cas9 1305 1310 1315 1320 EQAENIIHLFTLTNLGAPAA Cas9 EQAENIIHLFTLTNLGAPAA 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 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 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 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

5400 1325 1330 1335 1340 FKYFDTTIDRKRYTSTKEVL Cas9 1325 1330 1335 1340 FKYFDTTIDRKRYTSTKEVL Cas9 FKYFDTTIDRKRYTSTKEVL 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 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 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 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

5460 1345 1350 1355 1360 DATLIHQSITGLYETRIDLS Cas9 1345 1350 1355 1360 DATLIHQSITGLYETRIDLS Cas9 DATLIHQSITGLYETRIDLS 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 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 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 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

FseI 5520 1365 QLGGD 1 5 10 15 KRPAATKKAGQAKKK Cas9 nucleoplasmin NLS 1365 QLGGD (in frame with Cas9) KRPAATKKAGQAKKK Cas9 QLGGDKRPAATKKAGQAKKK 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 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 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 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

EcoRI 5580 K* bGH poly(A) signal K * nucleoplasmin NLS G G A G C T G A C A C G G A A G A T BGH-rev K* 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 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 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 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

5640 bGH poly(A) signal 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 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 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 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

5700 bGH poly(A) signal 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 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 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 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

5760 bGH poly(A) signal 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 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 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 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

NotI 5820 bGH poly(A) signal AAV2 ITR AAV2 ITR 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 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 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 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

5880 AAV2 ITR AAV2 ITR 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 c c c g g g c t t t g c c c g g g c g g c c t c 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 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

SbfIKasINarISfoIPluTI 5940 AAV2 ITR AAV2 ITR 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 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 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 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

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

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

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

6180 f1 ori c c c c g t c a a g c t c t a a a t c g g g g g c t c c c t t t a g g g t t c c g a t t t a g t g c t t t a c g g c a c g g g g c a g t t c g a g a t t t a g c c c c c g a g g g a a a t c c c a a g g c t a a a t c a c g a a a t g c c g t g

6240 f1 ori c t c g a c c c c a a a a a a c t t g a t t t g g g t g a t g g t t c a c g t a g t g g g c c a t c g c c c t g a t a g g a g c t g g g g t t t t t t g a a c t a a a c c c a c t a c c a a g t g c a t c a c c c g g t a g c g g g a c t a t c

6300 f1 ori G T G G A C T C T T G T T C C A A F1ori-F a c g g t t t t t c g c c c t t t g a c g t t g g a g t c c a c g t t c t t t a a t a g t g g a c t c t t g t t c c a a t g c c a a a a a g c g g g a a a c t g c a a c c t c a g g t g c a a g a a a t t a t c a c c t g a g a a c a a g g t t

PsiI 6360 f1 ori A C T G G F1ori-F a c t g g a a c a a c a c t c a a c c c t a t c t c g g g c t a t t c t t t t g a t t t a t a a g g g a t t t t g c c g t g a c c t t g t t g t g a g t t g g g a t a g a g c c c g a t a a g a a a a c t a a a t a t t c c c t a a a a c g g c

6420 f1 ori a t t t c g g c c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a t t t a a c g c g a a t t t t a a c t a a a g c c g g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t a a a t t g c g c t t a a a a t t g

6480 f1 ori A T G T T A G A C G A G A C T A C G G C pRS-marker a a a a t a t t a a c g t t t a c a a t t t t a t g g t g c a c t c t c a g t a c a a t c t g c t c t g a t g c c g c a t t t t a t a a t t g c a a a t g t t a a a a t a c c a c g t g a g a g t c a t g t t a g a c g a g a c t a c g g c g t

6540 t a g t t a a g c c a g c c c c g a c a c c c g c c a a c a c c c g c t g a c g c g c c c t g a c g g g c t t g t c t g a t c a a t t c g g t c g g g g c t g t g g g c g g t t g t g g g c g a c t g c g c g g g a c t g c c c g a a c a g a c

6600 C C G G G A G C T G C A T G T G T C A G A G G pGEX 3' c t c c c g g c a t c c g c t t a c a g a c a a g c t g t g a c c g t c t c c g g g a g c t g c a t g t g t c a g a g g g a g g g c c g t a g g c g a a t g t c t g t t c g a c a c t g g c a g a g g c c c t c g a c g t a c a c a g t c t c c

6660 C G G A G C A C T A T G C G G A T A A pBRforEco t t t t c a c c g t c a t c a c c g a a a c g c g c g a g a c g a a a g g g c c t c g t g a t a c g c c t a t t t t t a a a a a g t g g c a g t a g t g g c t t t g c g c g c t c t g c t t t c c c g g a g c a c t a t g c g g a t a a a a a t

6720 t a g g t t a a t g t c a t g a t a a t a a t g g t t t c t t a g a c g t c a g g t g g c a c t t t t c g g g g a a a t a t c c a a t t a c a g t a c t a t t a t t a c c a a a g a a t c t g c a g t c c a c c g t g a a a a g c c c c t t t a

6780 AmpR promoter g t g c g c g g a a c c c c t a t t t g t t t a t t t t t c t a a a t a c a t t c a a a t a t g t a t c c g c t c a t g c a c g c g c c t t g g g g a t a a a c a a a t a a a a a g a t t t a t g t a a g t t t a t a c a t a g g c g a g t a c

6840 1 signal sequence MSIQ AmpR promoter AmpR MSIQ a g a c a a t a a c c c t g a t a a a t g c t t c a a t a a t a t t g a a a a a g g a a g a g t a t g a g t a t t c a a t c t g t t a t t g g g a c t a t t t a c g a a g t t a t t a t a a c t t t t t c c t t c t c a t a c t c a t a a g t t

6900 5 10 15 20 signal sequence HFRVALIPFFAAFCLPVFA H AmpR HFRVALIPFFAAFCLPVFAH c a t t t c c g t g t c g c c c t t a t t c c c t t t t t t g c g g c a t t t t g c c t t c c t g t t t t t g c t c a c g t a a a g g c a c a g c g g g a a t a a g g g a a a a a a c g c c g t a a a a c g g a a g g a c a a a a a c g a g t g

6960 25 30 35 40 PETLVKVKDAEDQLGARVGY AmpR PETLVKVKDAEDQLGARVGY c c a g a a a c g c t g g t g a a a g t a a a a g a t g c t g a a g a t c a g t t g g g t g c a c g a g t g g g t t a c g g t c t t t g c g a c c a c t t t c a t t t t c t a c g a c t t c t a g t c a a c c c a c g t g c t c a c c c a a t g

7020 45 50 55 60 IELDLNSGKILESFRPEERF AmpR IELDLNSGKILESFRPEERF a t c g a a c t g g a t c t c a a c a g c g g t a a g a t c c t t g a g a g t t t t c g c c c c g a a g a a c g t t t t t a g c t t g a c c t a g a g t t g t c g c c a t t c t a g g a a c t c t c a a a a g c g g g g c t t c t t g c a a a a

7080 65 70 75 80 PMMSTFKVLLCGAVLSRIDA AmpR C G A T A C A C C G C G C C A T A A T A Amp-R PMMSTFKVLLCGAVLSRIDA c c a a t g a t g a g c a c t t t t a a a g t t c t g c t a t g t g g c g c g g t a t t a t c c c g t a t t g a c g c c g g t t a c t a c t c g t g a a a a t t t c a a g a c g a t a c a c c g c g c c a t a a t a g g g c a t a a c t g c g g

7140 85 90 95 100 GQEQLGRRIHYSQNDLVEYS AmpR GQEQLGRRIHYSQNDLVEYS g g g c a a g a g c a a c t c g g t c g c c g c a t a c a c t a t t c t c a g a a t g a c t t g g t t g a g t a c t c a c c c g t t c t c g t t g a g c c a g c g g c g t a t g t g a t a a g a g t c t t a c t g a a c c a a c t c a t g a g t

7200 105 110 115 120 PVTEKHLTDGMTVRELCSAA AmpR PVTEKHLTDGMTVRELCSAA c c a g t c a c a g a a a a g c a t c t t a c g g a t g g c a t g a c a g t a a g a g a a t t a t g c a g t g c t g c c g g t c a g t g t c t t t t c g t a g a a t g c c t a c c g t a c t g t c a t t c t c t t a a t a c g t c a c g a c g g

PvuI 7260 125 130 135 140 ITMSDNTAANLLLTTIGGPK AmpR ITMSDNTAANLLLTTIGGPK a t a a c c a t g a g t g a t a a c a c t g c g g c c a a c t t a c t t c t g a c a a c g a t c g g a g g a c c g a a g t a t t g g t a c t c a c t a t t g t g a c g c c g g t t g a a t g a a g a c t g t t g c t a g c c t c c t g g c t t c

7320 145 150 155 160 ELTAFLHNMGDHVTRLDRWE AmpR ELTAFLHNMGDHVTRLDRWE g a g c t a a c c g c t t t t t t g c a c a a c a t g g g g g a t c a t g t a a c t c g c c t t g a t c g t t g g g a a c t c g a t t g g c g a a a a a a c g t g t t g t a c c c c c t a g t a c a t t g a g c g g a a c t a g c a a c c c t t

7380 165 170 175 180 PELNEAIPNDERDTTMPVAM AmpR PELNEAIPNDERDTTMPVAM c c g g a g c t g a a t g a a g c c a t a c c a a a c g a c g a g c g t g a c a c c a c g a t g c c t g t a g c a a t g g g c c t c g a c t t a c t t c g g t a t g g t t t g c t g c t c g c a c t g t g g t g c t a c g g a c a t c g t t a c

FspI 7440 185 190 195 200 ATTLRKLLTGELLTLASRQQ AmpR ATTLRKLLTGELLTLASRQQ g c a a c a a c g t t g c g c a a a c t a t t a a c t g g c g a a c t a c t t a c t c t a g c t t c c c g g c a a c a a c g t t g t t g c a a c g c g t t t g a t a a t t g a c c g c t t g a t g a a t g a g a t c g a a g g g c c g t t g t t

7500 205 210 215 220 LIDWMEADKVAGPLLRSALP AmpR LIDWMEADKVAGPLLRSALP t t a a t a g a c t g g a t g g a g g c g g a t a a a g t t g c a g g a c c a c t t c t g c g c t c g g c c c t t c c g a a t t a t c t g a c c t a c c t c c g c c t a t t t c a a c g t c c t g g t g a a g a c g c g a g c c g g g a a g g c

SacII 7560 225 230 235 240 AGWFIADKSGAGERGSRGII AmpR AGWFIADKSGAGERGSRGII g c t g g c t g g t t t a t t g c t g a t a a a t c t g g a g c c g g t g a g c g t g g a a g c c g c g g t a t c a t t c g a c c g a c c a a a t a a c g a c t a t t t a g a c c t c g g c c a c t c g c a c c t t c g g c g c c a t a g t a a

7620 245 250 255 260 AALGPDGKPSRIVVIYTTGS AmpR AALGPDGKPSRIVVIYTTGS g c a g c a c t g g g g c c a g a t g g t a a g c c c t c c c g t a t c g t a g t t a t c t a c a c g a c g g g g a g t c g t c g t g a c c c c g g t c t a c c a t t c g g g a g g g c a t a g c a t c a a t a g a t g t g c t g c c c c t c a

7680 265 270 275 280 QATMDERNRQIAEIGASLIK AmpR QATMDERNRQIAEIGASLIK c a g g c a a c t a t g g a t g a a c g a a a t a g a c a g a t c g c t g a g a t a g g t g c c t c a c t g a t t a a g g t c c g t t g a t a c c t a c t t g c t t t a t c t g t c t a g c g a c t c t a t c c a c g g a g t g a c t a a t t c

7740 285 HW* AmpR HW* c a t t g g t a a c t g t c a g a c c a a g t t t a c t c a t a t a t a c t t t a g a t t g a t t t a a a a c t t c a t g t a a c c a t t g a c a g t c t g g t t c a a a t g a g t a t a t a t g a a a t c t a a c t a a a t t t t g a a g t a

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

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

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

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

8040 ori 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 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 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 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

8100 ori a a g a a c t c t g t a g c a c c g c c t a c a t a c c t c g c t c t g c t a a t c c t g t t a c c a g t g g c t g c t t t c t t g a g a c a t c g t g g c g g a t g t a t g g a g c g a g a c g a t t a g g a c a a t g g t c a c c g a c g a

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

8220 ori g c g c a g c g g t c g g g c t g a a c g g g g g g t t c g t g c a c a c a g c c c a g c t t g g a g c g a a c g a c c c g c g t c g c c a g c c c g a c t t g c c c c c c a a g c a c g t g t g t c g g g t c g a a c c t c g c t t g c t g g

8280 ori t a c a c c g a a c t g a g a t a c c t a c a g c g t g a g c t a t g a g a a a g c g c c a c g c t t c c c g a a g g g a t g t g g c t t g a c t c t a t g g a t g t c g c a c t c g a t a c t c t t t c g c g g t g c g a a g g g c t t c c c

8340 ori a g a a a g g c g g a c a g g t a t c c g g t a a g c g g c a g g g t c g g a a c a g g a g a g c g c a c g a g g g a g t c t t t c c g c c t g t c c a t a g g c c a t t c g c c g t c c c a g c c t t g t c c t c t c g c g t g c t c c c t c

8400 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F c t t c c a g g g g g a a a c g c c t g g t a t c t t t a t a g t c c t g t c g g g t t t c g c c a c c t c t g a c t t g a a g g t c c c c c t t t g c g g a c c a t a g a a a t a t c a g g a c a g c c c a a a g c g g t g g a g a c t g a a

8460 ori g a g c g t c g a t t t t t g t g a t g c t c g t c a g g g g g g c g g a g c c t a t g g a a a a a c g c c a g c a a c c t c g c a g c t a a a a a c a c t a c g a g c a g t c c c c c c g c c t c g g a t a c c t t t t t g c g g t c g t t g

AflIIIPciI 8509 g c g g c c t t t t t a c g g t t c c t g g c c t t t t g c t g g c c t t t t g c t c a c a t g t c g c c g g a a a a a t g c c a a g g a c c g g a a a a c g a c c g g a a a a c g a g t g t a c a

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.