Addgene pBK1313-AAV-EFSNC-dSaCas9-MECP2(204-310) Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pBK1313-AAV-EFSNC-dSaCas9-MECP2(204-310) Sequencing Result


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PciI (7326) pBR322ori-F (7171 .. 7190) SacII (6373) FspI (6215) PvuI (6069) Amp-R (5869 .. 5888) AatII (5519) ZraI (5517) pBRforEco (5460 .. 5478) pGEX 3' (5400 .. 5422) pRS-marker (5281 .. 5300) F1ori-F (5106 .. 5127) F1ori-R (4896 .. 4915) NotI (4587) HincII (4565) NdeI (4414) LKO.1 5' (4407 .. 4426) hU6-F (4236 .. 4256) KpnI (4234) Acc65I (4230) poly(A) signal SV40 NLS BmtI (4132) NheI (4128) BbsI (4060) Bsu36I (3979) PaqCI (3809) SpeI (3801) AAV2 ITR (alternate) MfeI (238) EcoNI (256) AgeI (399) NcoI (409) SV40 NLS SV40 NLS BspDI * - ClaI * (640) HindIII (1910) BspEI (2080) CsiI - SexAI * (2488) SphI (2777) FseI (3742) pBK1313-AAV-EFSNC-dSaCas9-MECP2(204-310) 7331 bp

Sequence View

60 AAV2 ITR (alternate) c c t g c a g g c a g 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 c c c g g g c g t c g g g c g a c c t t t g g a c g t c c g t c 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 g g g c c c g c a g c c c g c t g g a a a

120 AAV2 ITR (alternate) g g t c g c c 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 a g a g g g a g t g g c c a a c t c c a t c a c t c c a g c g g 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 t c t c c c t c a c c g g t t g a g g t a g t g a

180 AAV2 ITR (alternate) *SPRSLLPFQSRHG a g g g g t t c c t g c g g c c t c t a g c t a g g t c t t g a a a g g a g t g g g a a t t g g c t c c g g t g c c c g t c c c c a a g g a c g c c g g a g a t c g a t c c a g a a c t t t c c t c a c c c t t a a c c g a g g c c a c g g g c

MfeI 240 EF-1 α core promoter DTPLACMAWLGRSTPPLPRC t c a g t g g g c a g a g c g c a c a t c g c c c a c a g t c c c c g a g a a g t t g g g g g g a g g g g t c g g c a a a g t c a c c c g t c t c g c g t g t a g c g g g t g t c a g g g g c t c t t c a a c c c c c c t c c c c a g c c g t t

EcoNI 300 EF-1 α core promoter NIRHRSFTARPLSPFHHRTS t t g a t c c g g t g c c t a g a g a a g g t g g c g c g g g g t a a a c t g g g a a a g t g a t g t c g t g t a c t g a a c t a g g c c a c g g a t c t c t t c c a c c g c g c c c c a t t t g a c c c t t t c a c t a c a g c a c a t g a c

360 EF-1 α core promoter AGGKERPHPLVTYLHLLRRS g c t c c g c c t t t t t c c c g a g g g t g g g g g a g a a c c g t a t a t a a g t g c a g t a g t c g c c g t g a a c g a g g c g g a a a a a g g g c t c c c a c c c c c t c t t g g c a t a t a t t c a c g t c a t c a g c g g c a c t t

NcoIAgeI 420 1 DY EF-1 α core promoter 3xFLAG Kozak sequence MDY REKECRTQRWFVPGTGGHVV c g t t c t t t t t c g c a a c g g g t t t g c c g c c a g a a c a c a g g a c c g g t g c c a c c a t g g a c t a c a g c a a g a a a a a g c g t t g c c c a a a c g g c g g t c t t g t g t c c t g g c c a c g g t g g t a c c t g a t g t

480 5 10 15 20 KDHDGDYKDHDIDYKDDDDK 3xFLAG KDHDGDYKDHDIDYKDDDDK FVMVTIIFIMVYIVLIVIVL a a g a c c a t g a c g g t g a t t a t a a a g a t c a t g a c a t a g a t t a c a a g g a t g a c g a t g a c a a g g t t c t g g t a c t g c c a c t a a t a t t t c t a g t a c t g t a t c t a a t g t t c c t a c t g c t a c t g t t c c

540 GGS 130 PKKKRKV A 130 PKKKRKV GI SV40 NLS SV40 NLS GGSPKKKRKVAPKKKRKVGI TSRWLFLPFDGWLLLPLDTD g t g g a t c g c c a a a g a a a a a g a g g a a a g t c 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 c a c c t a g c g g t t t c t t t t t c t c c t t t c a g 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

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

BspDI*ClaI* 660 (in frame with SV40 NLS) VGYGIIDYETRDVIDAGVRL VGYGIIDYETRDVIDAGVRL HAVADDVVLCPVHDIGAHPQ t g g g c t a c g g c a t c a t c g a c t a c g a g a c a c g g g a c g t g a t c g a t g c c g g c g t g c g g c t g t a c c c g a t g c c g t a g t a g c t g a t g c t c t g t g c c c t g c a c t a g c t a c g g c c g c a c g c c g a c a

720 (in frame with SV40 NLS) FKEANVENNEGRRSKRGARR FKEANVENNEGRRSKRGARR EFLGVHFVVLAPPALSAGSP t c a a a g a g g c c a a c g t g g a a a a c a a c g a g g g c a g g c g g a g c a a g a g a g g c g c c a g a a g g c a g t t t c t c c g g t t g c a c c t t t t g t t g c t c c c g t c c g c c t c g t t c t c t c c g c g g t c t t c c g

780 (in frame with SV40 NLS) LKRRRRHRIQRVKKLLFDYN LKRRRRHRIQRVKKLLFDYN QLPPPPMSDLSHLLQQEVVV t g a a g c g g c g g a g g c g g c a t a g a a t c c a g a g a g t g a a g a a g c t g c t g t t c g a c t a c a a c c a c t t c g c c g c c t c c g c c g t a t c t t a g g t c t c t c a c t t c t t c g a c g a c a a g c t g a t g t t g g

840 (in frame with SV40 NLS) LLTDHSELSGINPYEARVKG LLTDHSELSGINPYEARVKG QQGVVALQAADVGVLGSHLA t g c t g a c c g a c c a c a g c g a g c t g a g c g g c a t c a a c c c c t a c g a g g c c a g a g t g a a g g g c c a c g a c t g g c t g g t g t c g c t c g a c t c g c c g t a g t t g g g g a t g c t c c g g t c t c a c t t c c c g g

900 (in frame with SV40 NLS) LSQKLSEEEFSAALLHLAKR LSQKLSEEEFSAALLHLAKR QALLQALFLERGGQQVQGLS t g a g c c a g a a g c t g a g c g a g g a a g a g t t c t c t g c c g c c c t g c t g c a c c t g g c c a a g a g a a a c t c g g t c t t c g a c t c g c t c c t t c t c a a g a g a c g g c g g g a c g a c g t g g a c c g g t t c t c t t

960 (in frame with SV40 NLS) RGVHNVNEVEEDTGNELSTK RGVHNVNEVEEDTGNELSTK SAHVVHVLHFLVGAVLQGGF g a g g c g t g c a c a a c g t g a a c g a g g t g g a a g a g g a c a c c g g c a a c g a g c t g t c c a c c a a a g c t c c g c a c g t g t t g c a c t t g c t c c a c c t t c t c c t g t g g c c g t t g c t c g a c a g g t g g t t t c

1020 (in frame with SV40 NLS) EQISRNSKALEEKYVAELQL EQISRNSKALEEKYVAELQL LLDAPVALGQFLFVHGFQLQ a g c a g a t c a g c c g g a a c a g c a a g g c c c t g g a a g a g a a a t a c g t g g c c g a a c t g c a g c t g g t c g t c t a g t c g g c c t t g t c g t t c c g g g a c c t t c t c t t t a t g c a c c g g c t t g a c g t c g a c c

1080 (in frame with SV40 NLS) ERLKKDGEVRGSINRFKTSD ERLKKDGEVRGSINRFKTSD FPQLFVAFHPAADVSELGAV a a c g g c t g a a g a a a g a c g g c g a a g t g c g g g g c a g c a t c a a c a g a t t c a a g a c c a g c g a c t t t g c c g a c t t c t t t c t g c c g c t t c a c g c c c c g t c g t a g t t g t c t a a g t t c t g g t c g c t g a

1140 (in frame with SV40 NLS) YVKEAKQLLKVQKAYHQLDQ YVKEAKQLLKVQKAYHQLDQ VHFFGFLQQLHLLGVVLQVL a c g t g a a a g a a g c c a a a c a g c t g c t g a a g g t g c a g a a g g c c t a c c a c c a g c t g g a c c a g a t g c a c t t t c t t c g g t t t g t c g a c g a c t t c c a c g t c t t c c g g a t g g t g g t c g a c c t g g t c t

1200 (in frame with SV40 NLS) SFIDTYIDLLETRRTYYEGP SFIDTYIDLLETRRTYYEGP AEDVGVDVQQFGPPGVILSR g c t t c a t c g a c a c c t a c a t c g a c c t g c t g g a a a c c c g g c g g a c c t a c t a t g a g g g a c c t g c g a a g t a g c t g t g g a t g t a g c t g g a c g a c c t t t g g g c c g c c t g g a t g a t a c t c c c t g g a c

1260 (in frame with SV40 NLS) GEGSPFGWKDIKEWYEMLMG GEGSPFGWKDIKEWYEMLMG ALAAGEAPLVDFFPVLHQHA g c g a g g g c a g c c c c t t c g g c t g g a a g g a c a t c a a a g a a t g g t a c g a g a t g c t g a t g g g c c c g c t c c c g t c g g g g a a g c c g a c c t t c c t g t a g t t t c t t a c c a t g c t c t a c g a c t a c c c g g

1320 (in frame with SV40 NLS) HCTYFPEELRSVKYAYNADL HCTYFPEELRSVKYAYNADL VAGVEGLFQPAHLVGVVGVQ a c t g c a c c t a c t t c c c c g a g g a a c t g c g g a g c g t g a a g t a c g c c t a c a a c g c c g a c c t g t t g a c g t g g a t g a a g g g g c t c c t t g a c g c c t c g c a c t t c a t g c g g a t g t t g c g g c t g g a c a

1380 (in frame with SV40 NLS) YNALNDLNNLVITRDENEKL YNALNDLNNLVITRDENEKL VVGQVVQVIEHDGPVLVLLQ a c a a c g c c c t g a a c g a c c t g a a c a a t c t c g t g a t c a c c a g g g a c g a g a a c g a g a a g c t g g t g t t g c g g g a c t t g c t g g a c t t g t t a g a g c a c t a g t g g t c c c t g c t c t t g c t c t t c g a c c

1440 (in frame with SV40 NLS) EYYEKFQIIENVFKQKKKPT EYYEKFQIIENVFKQKKKPT FIVLLELDDLVHELLLLLGG a a t a t t a c g a g a a g t t c c a g a t c a t c g a g a a c g t g t t c a a g c a g a a g a a g a a g c c c a c c c t t a t a a t g c t c t t c a a g g t c t a g t a g c t c t t g c a c a a g t t c g t c t t c t t c t t c g g g t g g g

1500 (in frame with SV40 NLS) LKQIAKEILVNEEDIKGYRV LKQIAKEILVNEEDIKGYRV QLLDGFFDEHVFLINLAVSH t g a a g c a g a t c g c c a a a g a a a t c c t c g t g a a c g a a g a g g a t a t t a a g g g c t a c a g a g t g a a c t t c g t c t a g c g g t t t c t t t a g g a g c a c t t g c t t c t c c t a t a a t t c c c g a t g t c t c a c t

1560 (in frame with SV40 NLS) TSTGKPEFTNLKVYHDIKDI TSTGKPEFTNLKVYHDIKDI GAGALGLEGVQLHVVVDLVN c c a g c a c c g g c a a g c c c g a g t t c a c c a a c c t g a a g g t g t a c c a c g a c a t c a a g g a c a t t a g g t c g t g g c c g t t c g g g c t c a a g t g g t t g g a c t t c c a c a t g g t g c t g t a g t t c c t g t a a t

1620 (in frame with SV40 NLS) TARKEIIENAELLDQIAKIL TARKEIIENAELLDQIAKIL GGPFLNNLVGLQQILNGLDQ c c g c c c g g a a a g a g a t t a t t g a g a a c g c c g a g c t g c t g g a t c a g a t t g c c a a g a t c c t g a g g c g g g c c t t t c t c t a a t a a c t c t t g c g g c t c g a c g a c c t a g t c t a a c g g t t c t a g g a c t

1680 (in frame with SV40 NLS) TIYQSSEDIQEELTNLNSEL TIYQSSEDIQEELTNLNSEL GDVLAALVDLFFQGIQVGLQ c c a t c t a c c a g a g c a g c g a g g a c a t c c a g g a a g a a c t g a c c a a t c t g a a c t c c g a g c t g a g g t a g a t g g t c t c g t c g c t c c t g t a g g t c c t t c t t g a c t g g t t a g a c t t g a g g c t c g a c t

1740 (in frame with SV40 NLS) TQEEIEQISNLKGYTGTHNL TQEEIEQISNLKGYTGTHNL *VPVWLRL GLFLDLLDRIQLAIGAGVVQ c c c a g g a a g a g a t c g a g c a g a t c t c t a a t c t g a a g g g c t a t a c c g g c a c c c a c a a c c t g a g g g t c c t t c t c t a g c t c g t c t a g a g a t t a g a c t t c c c g a t a t g g c c g t g g g t g t t g g a c t

1800 (in frame with SV40 NLS) SLKAINLILDELWHTNDNQI SLKAINLILDELWHTNDNQI RFAMLRIRSSSHCVLSLWIA AQLGDVQDQVLQPVGVVVLD g c c t g a a g g c c a t c a a c c t g a t c c t g g a c g a g c t g t g g c a c a c c a a c g a c a a c c a g a t c g c g g a c t t c c g g t a g t t g g a c t a g g a c c t g c t c g a c a c c g t g t g g t t g c t g t t g g t c t a g c

1860 (in frame with SV40 NLS) AIFNRLKLVPKKVDLSQQKE AIFNRLKLVPKKVDLSQQKE IKLRSFSTGLFTSRDWCFSI SDEVPQLQHGLLHVQGLLFL c t a t c t t c a a c c g g c t g a a g c t g g t g c c c a a g a a g g t g g a c c t g t c c c a g c a g a a a g a g a g a t a g a a g t t g g c c g a c t t c g a c c a c g g g t t c t t c c a c c t g g a c a g g g t c g t c t t t c t c t

HindIII 1920 (in frame with SV40 NLS) IPTTLVDDFILSPVVKRSFI IPTTLVDDFILSPVVKRSFI GVVRTSSKMRLGTTFLLKMW DGGGQHVVEDQAGDHLSAED t c c c c a c c a c c c t g g t g g a c g a c t t c a t c c t g a g c c c c g t c g t g a a g a g a a g c t t c a t c c a g g g g t g g t g g g a c c a c c t g c t g a a g t a g g a c t c g g g g c a g c a c t t c t c t t c g a a g t a g g

1980 (in frame with SV40 NLS) QSIKVINAIIKKYGLPNDII QSIKVINAIIKKYGLPNDII LMLTILAMMLFYPRGLSMMI LADFHDVGDDLLVAQGVVDN a g a g c a t c a a a g t g a t c a a c g c c a t c a t c a a g a a g t a c g g c c t g c c c a a c g a c a t c a t t a t c t c g t a g t t t c a c t a g t t g c g g t a g t a g t t c t t c a t g c c g g a c g g g t t g c t g t a g t a a t

2040 (in frame with SV40 NLS) IELAREKNSKDAQKMINEMQ IELAREKNSKDAQKMINEMQ SSARSFFELSAWFIILSICF DLQGALLVGLVGLFHDVLHL t c g a g c t g g c c c g c g a g a a g a a c t c c a a g g a c g c c c a g a a a a t g a t c a a c g a g a t g c a g a a g c t c g a c c g g g c g c t c t t c t t g a g g t t c c t g c g g g t c t t t t a c t a g t t g c t c t a c g t c t

BspEI 2100 (in frame with SV40 NLS) KRNRQTNERIEEIIRTTGKE KRNRQTNERIEEIIRTTGKE RFRCVLSRISSIMRVVPLSF LPVPLGVLPDLFDDPGGAFL a g c g g a a c c g g c a g a c c a a c g a g c g g a t c g a g g a a a t c a t c c g g a c c a c c g g c a a a g a g a t c g c c t t g g c c g t c t g g t t g c t c g c c t a g c t c c t t t a g t a g g c c t g g t g g c c g t t t c t c t

2160 (in frame with SV40 NLS) NAKYLIEKIKLHDMQEGKCL NAKYLIEKIKLHDMQEGKCL ALYRISFILSCSMCSPLHRY VGLVQDLLDLQVVHLFALAQ a c g c c a a g t a c c t g a t c g a g a a g a t c a a g c t g c a c g a c a t g c a g g a a g g c a a g t g c c t g t t g c g g t t c a t g g a c t a g c t c t t c t a g t t c g a c g t g c t g t a c g t c c t t c c g t t c a c g g a c a

2220 (in frame with SV40 NLS) YSLEAIPLEDLLNNPFNYEV YSLEAIPLEDLLNNPFNYEV LRSAM VAQFGDRQFIQQVVGEVILH a c a g c c t g g a a g c c a t c c c t c t g g a a g a t c t g c t g a a c a a c c c c t t c a a c t a t g a g g t g g t g t c g g a c c t t c g g t a g g g a g a c c t t c t a g a c g a c t t g t t g g g g a a g t t g a t a c t c c a c c

2280 (in frame with SV40 NLS) DHIIPRSVSFDNSFNNKVLV DHIIPRSVSFDNSFNNKVLV VVDDGSAHGEVVAEVVLHEH a c c a c a t c a t c c c c a g a a g c g t g t c c t t c g a c a a c a g c t t c a a c a a c a a g g t g c t c g t g a t g g t g t a g t a g g g g t c t t c g c a c a g g a a g c t g t t g t c g a a g t t g t t g t t c c a c g a g c a c t

2340 (in frame with SV40 NLS) KQEEASKKGNRTPFQYLSSS KQEEASKKGNRTPFQYLSSS LLFFGALLAVPGWELVQAAA a g c a g g a a g a a g c c a g c a a g a a g g g c a a c c g g a c c c c a t t c c a g t a c c t g a g c a g c a g c g t c g t c c t t c t t c g g t c g t t c t t c c c g t t g g c c t g g g g t a a g g t c a t g g a c t c g t c g t c g c

2400 (in frame with SV40 NLS) DSKISYETFKKHILNLAKGK DSKISYETFKKHILNLAKGK VALDAVFGELLVDQIQGLAL a c a g c a a g a t c a g c t a c g a a a c c t t c a a g a a g c a c a t c c t g a a t c t g g c c a a g g g c a a g g t g t c g t t c t a g t c g a t g c t t t g g a a g t t c t t c g t g t a g g a c t t a g a c c g g t t c c c g t t c c

2460 (in frame with SV40 NLS) GRISKTKKEYLLEERDINRF GRISKTKKEYLLEERDINRF ASDALGLFLIQQFFPVDVPE g c a g a a t c a g c a a g a c c a a g a a a g a g t a t c t g c t g g a a g a a c g g g a c a t c a a c a g g t t c t c g t c t t a g t c g t t c t g g t t c t t t c t c a t a g a c g a c c t t c t t g c c c t g t a g t t g t c c a a g a

SexAI*CsiI 2520 (in frame with SV40 NLS) SVQKDFINRNLVDTRYATRG SVQKDFINRNLVDTRYATRG GHLFVEDVPVQHIGSVGGSA c c g t g c a g a a a g a c t t c a t c a a c c g g a a c c t g g t g g a t a c c a g a t a c g c c a c c a g a g g c c g g c a c g t c t t t c t g a a g t a g t t g g c c t t g g a c c a c c t a t g g t c t a t g c g g t g g t c t c c g g

2580 (in frame with SV40 NLS) LMNLLRSYFRVNNLDVKVKS LMNLLRSYFRVNNLDVKVKS QHVQQPAVESHVVQVHFHLG t g a t g a a c c t g c t g c g g a g c t a c t t c a g a g t g a a c a a c c t g g a c g t g a a a g t g a a g t c c a a c t a c t t g g a c g a c g c c t c g a t g a a g t c t c a c t t g t t g g a c c t g c a c t t t c a c t t c a g g t

2640 (in frame with SV40 NLS) INGGFTSFLRRKWKFKKERN INGGFTSFLRRKWKFKKERN DIAAEGAKQPPLPLKLFLPV t c a a t g g c g g c t t c a c c a g c t t t c t g c g g c g g a a g t g g a a g t t t a a g a a a g a g c g g a a c a a g t t a c c g c c g a a g t g g t c g a a a g a c g c c g c c t t c a c c t t c a a a t t c t t t c t c g c c t t g t

2700 (in frame with SV40 NLS) KGYKHHAEDALIIANADFIF KGYKHHAEDALIIANADFIF LPVLVVGLVGQDNGVGIEDE a g g g g t a c a a g c a c c a c g c c g a g g a c g c c c t g a t c a t t g c c a a c g c c g a t t t c a t c t t c a t c c c c a t g t t c g t g g t g c g g c t c c t g c g g g a c t a g t a a c g g t t g c g g c t a a a g t a g a a g t

2760 (in frame with SV40 NLS) KEWKKLDKAKKVMENQMFEE KEWKKLDKAKKVMENQMFEE FLPLFQVLGFFHHFVLHELF a a g a g t g g a a g a a a c t g g a c a a g g c c a a a a a a g t g a t g g a a a a c c a g a t g t t c g a g g a a a t t c t c a c c t t c t t t g a c c t g t t c c g g t t t t t t c a c t a c c t t t t g g t c t a c a a g c t c c t t t

SphI 2820 (in frame with SV40 NLS) KQAESMPEIETEQEYKEIFI KQAESMPEIETEQEYKEIFI LLGLAHGLDFGLLLVFLDED a g c a g g c c g a g a g c a t g c c c g a g a t c g a a a c c g a g c a g g a g t a c a a a g a g a t c t t c a t c a t c g t c c g g c t c t c g t a c g g g c t c t a g c t t t g g c t c g t c c t c a t g t t t c t c t a g a a g t a g t

2880 (in frame with SV40 NLS) TPHQIKHIKDFKDYKYSHRV TPHQIKHIKDFKDYKYSHRV GGVLDLVNLVELVVLVAVPH c c c c c c a c c a g a t c a a g c a c a t t a a g g a c t t c a a g g a c t a c a a g t a c a g c c a c c g g g t g g g g g g g g t g g t c t a g t t c g t g t a a t t c c t g a a g t t c c t g a t g t t c a t g t c g g t g g c c c a c c

2940 (in frame with SV40 NLS) DKKPNRELINDTLYSTRKDD DKKPNRELINDTLYSTRKDD VLLRISLQNVVGQVGGPLVV a c a a g a a g c c t a a t a g a g a g c t g a t t a a c g a c a c c c t g t a c t c c a c c c g g a a g g a c g a c a t g t t c t t c g g a t t a t c t c t c g a c t a a t t g c t g t g g g a c a t g a g g t g g g c c t t c c t g c t g t

3000 (in frame with SV40 NLS) KGNTLIVNNLNGLYDKDNDK KGNTLIVNNLNGLYDKDNDK LAVGQDHVIQVAQVVLVIVL a g g g c a a c a c c c t g a t c g t g a a c a a t c t g a a c g g c c t g t a c g a c a a g g a c a a t g a c a a g c t c c c g t t g t g g g a c t a g c a c t t g t t a g a c t t g c c g g a c a t g c t g t t c c t g t t a c t g t t c g

3060 (in frame with SV40 NLS) LKKLINKSPEKLLMYHHDPQ LKKLINKSPEKLLMYHHDPQ QFLQDVLAGFLQQHVVVVGL t g a a a a a g c t g a t c a a c a a g a g c c c c g a a a a g c t g c t g a t g t a c c a c c a c g a c c c c c a g a a c t t t t t c g a c t a g t t g t t c t c g g g g c t t t t c g a c g a c t a c a t g g t g g t g c t g g g g g t c t

3120 (in frame with SV40 NLS) TYQKLKLIMEQYGDEKNPLY TYQKLKLIMEQYGDEKNPLY GVLFQLQNHFLVAVLLIGQV c c t a c c a g a a a c t g a a g c t g a t t a t g g a a c a g t a c g g c g a c g a g a a g a a t c c c c t g t a c a g g a t g g t c t t t g a c t t c g a c t a a t a c c t t g t c a t g c c g c t g c t c t t c t t a g g g g a c a t g t

3180 (in frame with SV40 NLS) KYYEETGNYLTKYSKKDNGP KYYEETGNYLTKYSKKDNGP LVVLFGPVVQGLVGFLVVAG a g t a c t a c g a g g a a a c c g g g a a c t a c c t g a c c a a g t a c t c c a a a a a g g a c a a c g g c c c c g t c a t g a t g c t c c t t t g g c c c t t g a t g g a c t g g t t c a t g a g g t t t t t c c t g t t g c c g g g g c

3240 (in frame with SV40 NLS) VIKKIKYYGNKLNAHLDITD VIKKIKYYGNKLNAHLDITD HDLLNLIVAVFQVGM t g a t c a a g a a g a t t a a g t a t t a c g g c a a c a a a c t g a a c g c c c a t c t g g a c a t c a c c g a c g a c t a g t t c t t c t a a t t c a t a a t g c c g t t g t t t g a c t t g c g g g t a g a c c t g t a g t g g c t g c

3300 (in frame with SV40 NLS) DYPNSRNKVVKLSLKPYRFD DYPNSRNKVVKLSLKPYRFD a c t a c c c c a a c a g c a g a a a c a a g g t c g t g a a g c t g t c c c t g a a g c c c t a c a g a t t c g a c g t g a t g g g g t t g t c g t c t t t g t t c c a g c a c t t c g a c a g g g a c t t c g g g a t g t c t a a g c t g c

3360 (in frame with SV40 NLS) VYLDNGVYKFVTVKNLDVIK VYLDNGVYKFVTVKNLDVIK t g t a c c t g g a c a a t g g c g t g t a c a a g t t c g t g a c c g t g a a g a a t c t g g a t g t g a t c a a a a a c a t g g a c c t g t t a c c g c a c a t g t t c a a g c a c t g g c a c t t c t t a g a c c t a c a c t a g t t t t

3420 (in frame with SV40 NLS) KENYYEVNSKCYEEAKKLKK KENYYEVNSKCYEEAKKLKK a a g a a a a c t a c t a c g a a g t g a a t a g c a a g t g c t a t g a g g a a g c t a a g a a g c t g a a g a a g a t t c t t t t g a t g a t g c t t c a c t t a t c g t t c a c g a t a c t c c t t c g a t t c t t c g a c t t c t t c t

3480 (in frame with SV40 NLS) ISNQAEFIASFYNNDLIKIN ISNQAEFIASFYNNDLIKIN t c a g c a a c c a g g c c g a g t t t a t c g c c t c c t t c t a c a a c a a c g a t c t g a t c a a g a t c a a c g a g t c g t t g g t c c g g c t c a a a t a g c g g a g g a a g a t g t t g t t g c t a g a c t a g t t c t a g t t g c

3540 (in frame with SV40 NLS) GELYRVIGVNNDLLNRIEVN GELYRVIGVNNDLLNRIEVN g c g a g c t g t a t a g a g t g a t c g g c g t g a a c a a c g a c c t g c t g a a c c g g a t c g a a g t g a a c a c g c t c g a c a t a t c t c a c t a g c c g c a c t t g t t g c t g g a c g a c t t g g c c t a g c t t c a c t t g t

3600 (in frame with SV40 NLS) MIDITYREYLENMNDKRPPR MIDITYREYLENMNDKRPPR t g a t c g a c a t c a c c t a c c g c g a g t a c c t g g a a a a c a t g a a c g a c a a g a g g c c c c c c a g g a a c t a g c t g t a g t g g a t g g c g c t c a t g g a c c t t t t g t a c t t g c t g t t c t c c g g g g g g t c c t

3660 (in frame with SV40 NLS) IIKTIASKTQSIKKYSTDIL IIKTIASKTQSIKKYSTDIL t c a t t a a g a c a a t c g c c t c c a a g a c c c a g a g c a t t a a g a a g t a c a g c a c a g a c a t t c t g g a g t a a t t c t g t t a g c g g a g g t t c t g g g t c t c g t a a t t c t t c a t g t c g t g t c t g t a a g a c c

3720 (in frame with SV40 NLS) GNLYEVKSKKHPQIIKKG 1 KR nucleoplasmin NLS GNLYEVKSKKHPQIIKKGKR g c a a c c t g t a t g a a g t g a a a t c t a a g a a g c a c c c t c a g a t c a t c a a a a a g g g c a a a a g g c c g t t g g a c a t a c t t c a c t t t a g a t t c t t c g t g g g a g t c t a g t a g t t t t t c c c g t t t t c c g

FseI 3780 5 10 15 PAATKKAGQAKKKK (in frame with nucleoplasmin NLS) GSGGGS nucleoplasmin NLS PAATKKAGQAKKKKGSGGGS c g g c g g c c a c g a a a a a g g c c g g c c a g g c a a a a a a g a a a a a g g g a t c c g g t g g a g g a a g t g g c c g c c g g t g c t t t t t c c g g c c g g t c c g t t t t t t c t t t t t c c c t a g g c c a c c t c c t t c a c

PaqCISpeI 3840 (in frame with nucleoplasmin NLS) GGSGSGGTSSEGVQVKRVLE GGSGSGGTSSEGVQVKRVLE g c g g g t c a g g g t c g g g t g g c a c t a g t t c g g a g g g t g t g c a g g t g a a a a g g g t c c t g g a g a c g c c c a g t c c c a g c c c a c c g t g a t c a a g c c t c c c a c a c g t c c a c t t t t c c c a g g a c c t c t

3900 (in frame with nucleoplasmin NLS) KSPGKLLVKMPFQTSPGGKA KSPGKLLVKMPFQTSPGGKA a a a g t c c t g g g a a g c t c c t t g t c a a g a t g c c t t t t c a a a c t t c g c c a g g g g g c a a g g c t g t t t c a g g a c c c t t c g a g g a a c a g t t c t a c g g a a a a g t t t g a a g c g g t c c c c c g t t c c g a c

3960 (in frame with nucleoplasmin NLS) EGGGATTSTQVMVIKRPGRK EGGGATTSTQVMVIKRPGRK a g g g g g g t g g g g c c a c c a c a t c c a c c c a g g t c a t g g t g a t c a a a c g c c c c g g c a g g a a g c t c c c c c c a c c c c g g t g g t g t a g g t g g g t c c a g t a c c a c t a g t t t g c g g g g c c g t c c t t c g

Bsu36I 4020 (in frame with nucleoplasmin NLS) RKAEADPQAIPKKRGRKPGS RKAEADPQAIPKKRGRKPGS g a a a a g c t g a g g c c g a c c c t c a g g c c a t t c c c a a g a a a c g g g g c c g a a a g c c g g g g a g t g c t t t t c g a c t c c g g c t g g g a g t c c g g t a a g g g t t c t t t g c c c c g g c t t t c g g c c c c t c a c

BbsI 4080 (in frame with nucleoplasmin NLS) VVAAAAAEAKKKAVKESSIR VVAAAAAEAKKKAVKESSIR t g g t g g c a g c c g c t g c c g c c g a g g c c a a a a a g a a a g c c g t g a a g g a g t c t t c t a t c c g a t a c c a c c g t c g g c g a c g g c g g c t c c g g t t t t t c t t t c g g c a c t t c c t c a g a a g a t a g g c t a

BmtINheI 4140 (in frame with nucleoplasmin NLS) SVQETVLPIKKRKTREASGG SVQETVLPIKKRKTREASGG c t g t g c a g g a g a c a g t a c t c c c c a t c a a g a a g c g c a a g a c c c g g g a g g c t a g c g g a g g t g g a c a c g t c c t c t g t c a t g a g g g g t a g t t c t t c g c g t t c t g g g c c c t c c g a t c g c c t c c a c

4200 GS 130 PKKKRKV GS* SV40 NLS poly(A) signal GSPKKKRKVGS* g t t c g c c a a a g a a g a a a c g g a a g g t g g g a t c c t g a a a t a a a a g a t c t t t g t t t t c a t t a g c a a g c g g t t t c t t c t t t g c c t t c c a c c c t a g g a c t t t a t t t t c t a g a a a c a a a a g t a a t c

Acc65IKpnI 4260 poly(A) signal 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 a t c t g t g t g t t g g t t t t t t g t g t g c g g c c g g t a c c 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 t a g a c a c a c a a c c a a a a a a c a c a c g c c g g c c a t g 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

4320 U6 promoter 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 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 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 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

4380 U6 promoter 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 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 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 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

NdeI 4440 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' 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 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 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 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

4500 U6 promoter 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 c g a a a c a c c g g a g a c c a c g g c a g g t 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 g c t t t g t g g c c t c t g g t g c c g t c c a

4560 Sa gRNA scaffold c t c a g t t t t a g t a c t c t g g a a a c a g a a t c t a c t a a a a c a a g g c a a a a t g c c g t g t t t a t c g a g t c a a a a t c a t g a g a c c t t t g t c t t a g a t g a t t t t g t t c c g t t t t a c g g c a c a a a t a g

NotIHincII 4620 Sa gRNA scaffold AAV2 ITR AAV2 ITR t c g t c a a c t t g t t g g c g a g a t t t t t 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 a g c a g t t g a a c a a c c g c t c t a a a a a 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

4680 AAV2 ITR AAV2 ITR 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 c c 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 g c g g

4740 AAV2 ITR AAV2 ITR 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 c g c c 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 g c g g

4800 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 a g c a 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 t c g t

4860 f1 ori 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 g c a g 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 c g t c

4920 f1 ori G A G G A A A G C G A A A G A A G G G A F1ori-R c g t g a c c g c t a c a c t t g c c a g c g c c t t a g c g c c c g c t c c t t t c g c t t t c t t c c c t t c c t t g c a c t g g c g a t g t g a a c g g t c g c g g a a t c g c g g g c g a g g a a a g c g a a a g a a g g g a a g g a a

4980 f1 ori 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 t t 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 c c a a

5040 f1 ori 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 a c g 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 g t g c

5100 f1 ori 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 t c t t 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 a g a a

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

5220 f1 ori t g a t t t a t a a g g g a t t t t g c c g a t t t c g g t c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a c t a a a t a t t c c c t a a a a c g g c t a a a g c c a g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t

5280 f1 ori 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 c a g 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 a g t c

5340 A T G T T A G A C G A G A C T A C G G C pRS-marker 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 c t g a 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 g a c t

5400 C pGEX 3' 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 t c t c 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 a g a g

5460 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 pBRforEco 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 g 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 t c c c

ZraIAatII 5520 G G A G C A C T A T G C G G A T A A pBRforEco 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 c g t 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 g c a g

5580 AmpR promoter 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 t a c a 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 a t g t

5640 AmpR promoter 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 g a a a 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 c t t t

5700 1 5 10 15 signal sequence MSIQHFRVALIPFFAAF AmpR promoter AmpR MSIQHFRVALIPFFAAF 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 a 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 g t a a

5760 20 signal sequence CLPVFA 25 30 35 HPETLVKVKDAEDQ AmpR CLPVFAHPETLVKVKDAEDQ 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 a t c 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 t a g t

5820 40 45 50 55 LGARVGYIELDLNSGKILES AmpR LGARVGYIELDLNSGKILES 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 a g a g 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 t c t c

5880 60 65 70 75 FRPEERFPMMSTFKVLLCGA AmpR C G A T A C A C C G C G Amp-R FRPEERFPMMSTFKVLLCGA 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 g c g 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 c g c g

5940 80 85 90 95 VLSRIDAGQEQLGRRIHYSQ AmpR C C A T A A T A Amp-R VLSRIDAGQEQLGRRIHYSQ 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 t c a 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 g a g t

6000 100 105 110 115 NDLVEYSPVTEKHLTDGMTV AmpR NDLVEYSPVTEKHLTDGMTV 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 c 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 g t c a

6060 120 125 130 135 RELCSAAITMSDNTAANLLL AmpR RELCSAAITMSDNTAANLLL 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 t t c t 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 a a g a

PvuI 6120 140 145 150 155 TTIGGPKELTAFLHNMGDHV AmpR TTIGGPKELTAFLHNMGDHV 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 t g t 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 t a c a

6180 160 165 170 175 TRLDRWEPELNEAIPNDERD AmpR TRLDRWEPELNEAIPNDERD 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 g t g 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 c a c t

FspI 6240 180 185 190 195 TTMPVAMATTLRKLLTGELL AmpR TTMPVAMATTLRKLLTGELL 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 t 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 a t g a

6300 200 205 210 215 TLASRQQLIDWMEADKVAGP AmpR TLASRQQLIDWMEADKVAGP 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 g a c c 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 c t g g

6360 220 225 230 235 LLRSALPAGWFIADKSGAGE AmpR LLRSALPAGWFIADKSGAGE 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 g t g a 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 c a c t

SacII 6420 240 245 250 255 RGSRGIIAALGPDGKPSRIV AmpR RGSRGIIAALGPDGKPSRIV 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 t c g 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 a g c a

6480 260 265 270 275 VIYTTGSQATMDERNRQIAE AmpR VIYTTGSQATMDERNRQIAE 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 c t g 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 g a c t

6540 280 285 IGASLIKHW* AmpR IGASLIKHW* 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 t 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 a t g a

6600 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 t 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 a a c t

6660 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 c c g 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 g g c a

6720 ori 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 t 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 a c g t

6780 ori 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 c t c 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 g a g a

6840 ori 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 t c t t c t a g t g t 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 a g a a g a t c a c a t

6900 ori 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 t g c 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 a c g a

6960 ori 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 t c 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 t g a g

7020 ori 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 c a c a 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 g t g t

7080 ori 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 a g a 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 c t c t

7140 ori 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 t c g g 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 a g c c

7200 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F 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 c t g t 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 g a c a

7260 ori 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 g a g 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 c c t c

7320 ori 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 t t t 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 g a a a

PciI 7331 t g c t c a c a t g t 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.