Addgene SlugCas9-HF - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: SlugCas9-HF


Map View

PciI (7445) pBR322ori-F (7290 .. 7309) FspI (6334) PvuI (6188) Amp-R (5988 .. 6007) pBRforEco (5579 .. 5597) pGEX 3' (5519 .. 5541) pRS-marker (5400 .. 5419) F1ori-F (5225 .. 5246) F1ori-R (5015 .. 5034) PluTI (4858) SfoI (4856) NarI (4855) KasI (4854) EagI - NotI (4706) LKO.1 5' (4526 .. 4545) hU6-F (4355 .. 4375) KpnI (4352) Acc65I (4348) SphI (4340) bGH poly(A) signal BGH-rev (4134 .. 4151) EcoRI (4110) AflII (4105) BamHI (4020) FseI (4000) BbvCI (3946) XbaI (137) SpeI (168) SnaBI (509) CMV-F (688 .. 708) Kozak sequence SV40 NLS AleI (852) BspDI - ClaI (895) BsmI (988) EcoNI (1110) PspOMI (1451) ApaI (1455) KflI (1987) NsiI (2013) BspQI - SapI (2023) PmeI (2627) StuI (2799) BsrGI (3172) XcmI (3593) SlugCas9-HF 7450 bp

Sequence View

5ʹ 3ʹ 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

SpeIXbaI 180 AAV2 ITR (alternate) CMV enhancer a g g g g t t c c t g c g g c c t c t a g a c t c g a g g c g t t g a c a t t g a t t a t t g a c t a g t t a t t a a t t c c c c a a g g a c g c c g g a g a t c t g a g c t c c g c a a c t g t a a c t a a t a a c t g a t c a a t a a t t a

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

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

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

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

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

SnaBI 540 CMV enhancer CMV promoter g g g a c t t t c c t a c t t g g c a g t a c a t c t a c g t a t t a g t c a t c g c t a t t a c c a t g g t g a t g c c c c t g a a a g g a t g a a c c g t c a t g t a g a t g c a t a a t c a g t a g c g a t a a t g g t a c c a c t a c g

600 CMV promoter g g t t t t g g c a g t a c a t c a a t g g g c g t g g a t a g c g g t t t g a c t c a c g g g g a t t t c c a a g t c c c a a a a c c g t c a t g t a g t t a c c c g c a c c t a t c g c c a a a c t g a g t g c c c c t a a a g g t t c a g

660 CMV promoter t c c a c c c c a t t g a c g t c a a t g g g a g t t t g t t t t g g c a c c a a a a t c a a c g g g a c t t t c c a a a g g t g g g g t a a c t g c a g t t a c c c t c a a a c a a a a c c g t g g t t t t a g t t g c c c t g a a a g g t t

720 CMV promoter C G C A A A T G G G C G G T A G G C G T G CMV-F MGGRRVRWE a a t g t c g t a a c a a c t c c g c c c c a t t g a c g c a a a t g g g c g g t a g g c g t g t a c g g t g g g a g g t t a c a g c a t t g t t g a g g c g g g g t a a c t g c g t t t a c c c g c c a t c c g c a c a t g c c a c c c t c c

780 PKKK CMV promoter Kozak sequence SV40 NLS VYISRALWLTTGATMAPKKK *SGTGGHGWLLL t c t a t a t a a g c a g a g c t c t c t g g c t a a c t a c c g g t g c c a c c a t g g c c c c a a a g a a g a a g c a g a t a t a t t c g t c t c g a g a g a c c g a t t g a t g g c c a c g g t g g t a c c g g g g t t t c t t c t t c g

840 130 RKV (in frame with SV40 NLS) GIHGVPAANQKFILGLD SV40 NLS RKVGIHGVPAANQKFILGLD PLDTDVSDWCGVLFEYQSQV 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 a a c c a a a a a t t c a t a c t g g g a c t g g a c a 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 t t g g t t t t t a a g t a t g a c c c t g a c c t g t

BspDIClaIAleI 900 (in frame with SV40 NLS) IGITSVGYGLIDYETKNIID IGITSVGYGLIDYETKNIID DSDGAHAVAQDVVLCLIDDI t c g g a a t c a c c a g c g t g g g c t a c g g c c t g a t c g a c t a c g a g a c a a a g a a t a t c a t c g a t g a g c c t t a g t g g t c g c a c c c g a t g c c g g a c t a g c t g a t g c t c t g t t t c t t a t a g t a g c t a c

960 (in frame with SV40 NLS) AGVRLFPEANVENNEGRRSK AGVRLFPEANVENNEGRRSK GANSQEGLGVHFVVLSSPGF c c g g c g t t a g a c t g t t c c c c 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 a a g a a g g t c c a a a c g g c c g c a a t c t g a c a a g g g g 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 t t c t t c c a g g t t t g

BsmI 1020 (in frame with SV40 NLS) RGSRRLKRRRIHRLERVKKL RGSRRLKRRRIHRLERVKKL TSASSQLSAANVSKFPHLLQ g t g g a a g c a g a a g a c t g a a g c g a c g c c g c a t t c a c a g a c t t g a a c g g g t g a a g a a g c t g c c a c c t t c g t c t t c t g a c t t c g c t g c g g c g t a a g t g t c t g a a c t t g c c c a c t t c t t c g a c g

1080 (in frame with SV40 NLS) LEDYNLLDQSQIPQSTNPYA LEDYNLLDQSQIPQSTNPYA ELIIIQQILGLNRLRCVGVG t c g a g g a t t a t a a t c t g c t g g a t c a g t c c c a g a t t c c t c a g t c t a c a a a c c c c t a c g c c a a g c t c c t a a t a t t a g a c g a c c t a g t c a g g g t c t a a g g a g t c a g a t g t t t g g g g a t g c g g t

EcoNI 1140 (in frame with SV40 NLS) IRVKGLSEALSKDELVIALL IRVKGLSEALSKDELVIALL DAHLAQRFGQALVFEHNGQE t c c g c g t g a a g g g c c t g t c t g a a g c c c t g a g c a a g g a c g a a c t c g t g a t t g c c c t g c t c c a g g c g c a c t t c c c g g a c a g a c t t c g g g a c t c g t t c c t g c t t g a g c a c t a a c g g g a c g a g g

1200 (in frame with SV40 NLS) HIAKRRGIHKIDVIDSNDDV HIAKRRGIHKIDVIDSNDDV MDGLSSADVLDVHDVAVVVH a t a t c g c c a a g a g a a g a g g c a t c c a c a a g a t c g a c g t g a t c g a c a g c a a c g a c g a c g t g g t a t a g c g g t t c t c t t c t c c g t a g g t g t t c t a g c t g c a c t a g c t g t c g t t g c t g c t g c a c c

1260 (in frame with SV40 NLS) GNELSTKEQLNKNSKLLKDK GNELSTKEQLNKNSKLLKDK PVLEAGLFLQILVALQQFVF g g a a c g a g c t c a g c a c c a a g g a a c a g c t g a a t a a g a a c a g c a a g c t g c t g a a a g a c a a a t c c t t g c t c g a g t c g t g g t t c c t t g t c g a c t t a t t c t t g t c g t t c g a c g a c t t t c t g t t t a

1320 (in frame with SV40 NLS) FVCQIQLERMNEGQVRGEKN FVCQIQLERMNEGQVRGEKN KHALDLQFSHILALHPSLFV t t g t g t g c c a g a t c c a g c t g g a a a g a a t g a a t g a g g g c c a g g t g c g g g g a g a g a a a a a c c a a c a c a c g g t c t a g g t c g a c c t t t c t t a c t t a c t c c c g g t c c a c g c c c c t c t c t t t t t g g

1380 (in frame with SV40 NLS) RFKTADIIKEIIQLLNVQKN RFKTADIIKEIIQLLNVQKN PELGSIDDLFDDLQQIHLLV g g t t c a a g a c c g c t g a t a t c a t c a a g g a a a t c a t c c a g c t g c t g a a t g t g c a g a a g a a c t c c a a g t t c t g g c g a c t a t a g t a g t t c c t t t a g t a g g t c g a c g a c t t a c a c g t c t t c t t g a

1440 (in frame with SV40 NLS) FHQLDENFINKYIELVEMRR FHQLDENFINKYIELVEMRR EVLQVLVEDVLVDFQNLHSP t c c a c c a g c t g g a c g a g a a c t t c a t c a a c a a g t a c a t c g a a c t g g t t g a g a t g a g a c g g g a g g t g g t c g a c c t g c t c t t g a a g t a g t t g t t c a t g t a g c t t g a c c a a c t c t a c t c t g c c c

PspOMIApaI 1500 (in frame with SV40 NLS) EYFEGPGKGSPYGWEGDPKA EYFEGPGKGSPYGWEGDPKA FVELAGALATWIAPFAVRLS a a t a c t t c g a g g g c c c c g g c a a g g g c a g t c c a t a t g g c t g g g a a g g c g a c c c t a a g g c t t t t a t g a a g c t c c c g g g g c c g t t c c c g t c a g g t a t a c c g a c c c t t c c g c t g g g a t t c c g a a

1560 (in frame with SV40 NLS) WYETLMGHCTYFPDELASVK WYETLMGHCTYFPDELASVK PVLCQHAVAGVEWILQSAHF g g t a c g a g a c a c t g a t g g g c c a c t g c a c c t a c t t c c c a g a t g a g c t g g c t a g c g t g a a a t c c a t g c t c t g t g a c t a c c c g g t g a c g t g g a t g a a g g g t c t a c t c g a c c g a t c g c a c t t t a

1620 (in frame with SV40 NLS) YAYSADLFNALNDLNNLVIQ YAYSADLFNALNDLNNLVIQ VGVTGVQEVSQVVQVVQDDL a c g c c t a c a g t g c c g a c c t g t t c a a c g c t c t g a a c g a c c t g a a c a a c c t g g t c a t c c a a a t g c g g a t g t c a c g g c t g g a c a a g t t g c g a g a c t t g c t g g a c t t g t t g g a c c a g t a g g t t t

1680 (in frame with SV40 NLS) RDGLSKLEYHEKYHIIENVF RDGLSKLEYHEKYHIIENVF SISQRLELIM g a g a t g g a c t g t c t a a g c t c g a g t a t c a t g a g a a g t a t c a c a t c a t c g a g a a c g t g t t c a c t c t a c c t g a c a g a t t c g a g c t c a t a g t a c t c t t c a t a g t g t a g t a g c t c t t g c a c a a g t

1740 (in frame with SV40 NLS) KQKKKPTLKQIANEINVNPE KQKKKPTLKQIANEINVNPE a g c a g a a g a a g a a a c c t a c a c t g a a g c a g a t c g c c a a t g a g a t c a a t g t c a a c c c t g a a g t c g t c t t c t t c t t t g g a t g t g a c t t c g t c t a g c g g t t a c t c t a g t t a c a g t t g g g a c t t c

1800 (in frame with SV40 NLS) DIKGYRITKSGKPQFTEFKL DIKGYRITKSGKPQFTEFKL a t a t c a a g g g c t a c a g a a t c a c a a a g t c t g g c a a g c c c c a g t t t a c c g a g t t t a a g c t c t t a t a g t t c c c g a t g t c t t a g t g t t t c a g a c c g t t c g g g g t c a a a t g g c t c a a a t t c g a g a

1860 (in frame with SV40 NLS) YHDLKSVLFDQSILENEDVL YHDLKSVLFDQSILENEDVL a c c a c g a c c t g a a a a g c g t g c t g t t t g a c c a g a g c a t c c t g g a g a a c g a a g a c g t g c t g g t g g t g c t g g a c t t t t c g c a c g a c a a a c t g g t c t c g t a g g a c c t c t t g c t t c t g c a c g a c c

1920 (in frame with SV40 NLS) DQIAEILTIYQDKDSIKSKL DQIAEILTIYQDKDSIKSKL a c c a g a t c g c t g a g a t c c t g a c c a t c t a c c a g g a c a a g g a t a g c a t c a a a t c t a a g c t g a t g g t c t a g c g a c t c t a g g a c t g g t a g a t g g t c c t g t t c c t a t c g t a g t t t a g a t t c g a c t

1980 (in frame with SV40 NLS) TELDILLNEEDKENIAQLTG TELDILLNEEDKENIAQLTG c g g a a c t g g a c a t c c t g c t g a a c g a g g a a g a t a a g g a a a a c a t c g c c c a g c t g a c t g g c t g c c t t g a c c t g t a g g a c g a c t t g c t c c t t c t a t t c c t t t t g t a g c g g g t c g a c t g a c c g a

BspQISapINsiIKflI 2040 (in frame with SV40 NLS) YTGTHRLSLKCIRLVLEEQW YTGTHRLSLKCIRLVLEEQW a c a c c g g g a c c c a c c g g c t c a g c c t g a a a t g c a t c c g g c t g g t c c t g g a a g a g c a g t g g t t g t g g c c c t g g g t g g c c g a g t c g g a c t t t a c g t a g g c c g a c c a g g a c c t t c t c g t c a c c a

2100 (in frame with SV40 NLS) YSSRAQMEIFAHLNIKPKKI YSSRAQMEIFAHLNIKPKKI a t t c t a g c c g g g c t c a g a t g g a a a t c t t c g c c c a c c t g a a c a t t a a g c c t a a g a a g a t c a t a a g a t c g g c c c g a g t c t a c c t t t a g a a g c g g g t g g a c t t g t a a t t c g g a t t c t t c t a g t

2160 (in frame with SV40 NLS) NLTAANKIPKAMIDEFILSP NLTAANKIPKAMIDEFILSP a c c t g a c a g c c g c c a a c a a g a t c c c g a a g g c t a t g a t c g a c g a g t t c a t c c t g a g c c c t g t g g a c t g t c g g c g g t t g t t c t a g g g c t t c c g a t a c t a g c t g c t c a a g t a g g a c t c g g g a c

2220 (in frame with SV40 NLS) VVKRTFGQAINLINKIIEKY VVKRTFGQAINLINKIIEKY t g g t g a a g a g g a c c t t c g g c c a g g c c a t t a a c c t t a t t a a c a a g a t c a t a g a a a a g t a c g a c c a c t t c t c c t g g a a g c c g g t c c g g t a a t t g g a a t a a t t g t t c t a g t a t c t t t t c a t g c

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

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

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

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

2520 (in frame with SV40 NLS) NPNHYEVDHIIPRSVSFDNS NPNHYEVDHIIPRSVSFDNS a c c c c a a c c a c t a c g a g g t g g a c c a c a t c a t t c c a a g a t c t g t g t c c t t c g a c a a c t c t t t g g g g t t g g t g a t g c t c c a c c t g g t g t a g t a a g g t t c t a g a c a c a g g a a g c t g t t g a g a a

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

PmeI 2640 (in frame with SV40 NLS) YQYFNSGKSKLSYNQFKQHI YQYFNSGKSKLSYNQFKQHI a c c a g t a c t t t a a c a g c g g c a a g t c c a a g c t c t c t t a c a a c c a g t t t a a a c a a c a c a t c c t g g t c a t g a a a t t g t c g c c g t t c a g g t t c g a g a g a a t g t t g g t c a a a t t t g t t g t g t a g g

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

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

StuI 2820 (in frame with SV40 NLS) TRYATAELTNYLKAYFSANN TRYATAELTNYLKAYFSANN c c c g g t a c g c c a c a g c c g a g c t g a c a a a c t a c c t g a a g g c c t a c t t c a g c g c c a a c a a t a g g g c c a t g c g g t g t c g g c t c g a c t g t t t g a t g g a c t t c c g g a t g a a g t c g c g g t t g t t a t

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

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

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

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

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

BsrGI 3180 (in frame with SV40 NLS) YSHRVDKKPNRQLINDTLYS YSHRVDKKPNRQLINDTLYS a c a g c c a c a g a g t g g a c a a g a a g c c t a a c a g a c a g c t g a t c a a c g a t a c a c t g t a c a g c a t g t c g g t g t c t c a c c t g t t c t t c g g a t t g t c t g t c g a c t a g t t g c t a t g t g a c a t g t c g t

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

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

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

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

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

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

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

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

3720 (in frame with SV40 NLS) LGKAIDKNAKFIASFYKNDL LGKAIDKNAKFIASFYKNDL t g g g c a a g g c c a t c g a c a a a a a c g c c a a g t t t a t c g c t a g c t t c t a c a a g a a t g a t c t g a a c c c g t t c c g g t a g c t g t t t t t g c g g t t c a a a t a g c g a t c g a a g a t g t t c t t a c t a g a c t

3780 (in frame with SV40 NLS) IKLDGEIYKIIGVNSDTRNM IKLDGEIYKIIGVNSDTRNM t c a a g c t g g a c g g c g a g a t c t a c a a g a t c a t c g g c g t g a a t a g c g a c a c c a g a a a c a t g a a g t t c g a c c t g c c g c t c t a g a t g t t c t a g t a g c c g c a c t t a t c g c t g t g g t c t t t g t a c t

3840 (in frame with SV40 NLS) IELDLPDIRYKEYCELNNIK IELDLPDIRYKEYCELNNIK t c g a a c t g g a t c t g c c t g a c a t c a g a t a c a a a g a a t a c t g c g a g c t g a a c a a t a t c a a g g a g c t t g a c c t a g a c g g a c t g t a g t c t a t g t t t c t t a t g a c g c t c g a c t t g t t a t a g t t c c

3900 (in frame with SV40 NLS) GEPRIKKTIGKKVNSIEKLT GEPRIKKTIGKKVNSIEKLT g c g a a c c t a g a a t c a a a a a g a c c a t c g g c a a a a a g g t g a a t a g c a t c g a a a a a c t g a c a a c g c t t g g a t c t t a g t t t t t c t g g t a g c c g t t t t t c c a c t t a t c g t a g c t t t t t g a c t g t t

BbvCI 3960 (in frame with SV40 NLS) TDVLGNVFTNTQYTKPQLLF TDVLGNVFTNTQYTKPQLLF c c g a c g t g c t g g g c a a c g t g t t c a c c a a c a c c c a g t a c a c a a a a c c t c a g c t g c t g t t c a g g c t g c a c g a c c c g t t g c a c a a g t g g t t g t g g g t c a t g t g t t t t g g a g t c g a c g a c a a g t

FseI 4020 KRGN 1 5 10 15 KRPAATKKAGQAKKKK nucleoplasmin NLS KRGNKRPAATKKAGQAKKKK a g c g a g g a a a t 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 a g g t c g c t c c t t t a 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 t c c

BamHI 4080 GS 1 5 10 15 YPYDVPDYAYPYDVPDYA 3xHA GSYPYDVPDYAYPYDVPDYA g a t c c t a c c c a t a c g a t g t t c c a g a t t a c g c t t a c c c a t a c g a t g t t c c a g a t t a c g c t t c t a g g a t g g g t a t g c t a c a a g g t c t a a t g c g a a t g g g t a t g c t a c a a g g t c t a a t g c g a a

EcoRIAflII 4140 20 25 YPYDVPDYA * 3xHA bGH poly(A) signal G G A G C T G BGH-rev YPYDVPDYA* a c c c a t a c g a t g t t c c a g a t t a c g c t 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 g g t a t g c t a c a a g g t c t a a t g c g a 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

4200 bGH poly(A) signal A C A C G G A A G A T BGH-rev 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 c t 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 g a

4260 bGH poly(A) signal 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 c t 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 g a

4320 bGH poly(A) signal 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 t g 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 a c

SphIAcc65IKpnI 4380 bGH 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 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 g t a c c 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 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 c a t g g 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

4440 U6 promoter 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 a 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 t

4500 U6 promoter 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 c 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 g

4560 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 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 t 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 a

4620 U6 promoter 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 c 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 g

4680 Sa gRNA scaffold 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 t 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 a

EagINotI 4740 Sa gRNA scaffold AAV2 ITR AAV2 ITR 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 c 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 g

4800 AAV2 ITR AAV2 ITR 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 c 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 g

KasINarISfoIPluTI 4860 AAV2 ITR AAV2 ITR 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 t 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 a

4920 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 t

4980 f1 ori 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 c 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 g

5040 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 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 c t t t 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 g g a a a

5100 f1 ori 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 c 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 g

5160 f1 ori 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 t 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 a

5220 f1 ori 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 t 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 a

5280 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 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 c t t t t 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 g a a a a

5340 f1 ori 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 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 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

5400 f1 ori A pRS-marker 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 a

5460 T G T T A G A C G A G A C T A C G G C pRS-marker 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 c 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 g

5520 C C pGEX 3' 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 c 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 g

5580 G G G A G C T G C A T G T G T C A G A G G pGEX 3' C G pBRforEco 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 c 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 g

5640 G A G C A C T A T G C G G A T A A pBRforEco 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 a 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 t

5700 AmpR promoter 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 a

5760 AmpR promoter 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 t

5820 1 5 10 15 signal sequence MSIQHFRVALIPFFAAF AmpR promoter AmpR MSIQHFRVALIPFFAAF 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 a

5880 20 signal sequence CLPVFA 25 30 35 HPETLVKVKDAEDQ AmpR CLPVFAHPETLVKVKDAEDQ 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 g 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 c

5940 40 45 50 55 LGARVGYIELDLNSGKILES AmpR LGARVGYIELDLNSGKILES 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 t 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 a

6000 60 65 70 75 FRPEERFPMMSTFKVLLCGA AmpR C G A T A C A C C G C G C Amp-R FRPEERFPMMSTFKVLLCGA 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 g 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 c

6060 80 85 90 95 VLSRIDAGQEQLGRRIHYSQ AmpR C A T A A T A Amp-R VLSRIDAGQEQLGRRIHYSQ 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 g 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 c

6120 100 105 110 115 NDLVEYSPVTEKHLTDGMTV AmpR NDLVEYSPVTEKHLTDGMTV 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 a 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 t

6180 120 125 130 135 RELCSAAITMSDNTAANLLL AmpR RELCSAAITMSDNTAANLLL 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 g 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 c

PvuI 6240 140 145 150 155 TTIGGPKELTAFLHNMGDHV AmpR TTIGGPKELTAFLHNMGDHV 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 a 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 t

6300 160 165 170 175 TRLDRWEPELNEAIPNDERD AmpR TRLDRWEPELNEAIPNDERD 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 c 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 g

FspI 6360 180 185 190 195 TTMPVAMATTLRKLLTGELL AmpR TTMPVAMATTLRKLLTGELL 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 t 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 a

6420 200 205 210 215 TLASRQQLIDWMEADKVAGP AmpR TLASRQQLIDWMEADKVAGP 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 t

6480 220 225 230 235 LLRSALPAGWFIADKSGAGE AmpR LLRSALPAGWFIADKSGAGE 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 g 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 c

6540 240 245 250 255 RGSRGIIAALGPDGKPSRIV AmpR RGSRGIIAALGPDGKPSRIV 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 a 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 t

6600 260 265 270 275 VIYTTGSQATMDERNRQIAE AmpR VIYTTGSQATMDERNRQIAE 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 g 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 c

6660 280 285 IGASLIKHW* AmpR IGASLIKHW* 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 t 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 a

6720 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 t 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 a

6780 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 t

6840 ori 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 a 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 t

6900 ori 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 a

6960 ori 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 t g t a g 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 a c a t c

7020 ori 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 a 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 t

7080 ori 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 a 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 t

7140 ori 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 g 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 c

7200 ori 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 a 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 t

7260 ori 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 a 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 t

7320 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F 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 c 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 g

7380 ori 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 c 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 g

7440 ori 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 t 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 a

PciI 3ʹ 5ʹ 7450 g c t c a c a t g t 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.