pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro

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AmpR promoter SspI (10,966) PciI (9269) SfiI (8847) AanI (8618) AanI (8562) KpnI (8227) Acc65I (8223) HincII (7628) SalI (7626) SexAI * (7587) BstEII (7154) RsrII (7136) BsiWI (7076) PflFI - Tth111I (7062) BamHI (6954) NotI (6946) FseI (6925) nucleoplasmin NLS BsaBI * (6291) MluI (1) NruI * (612) MfeI (964) BbvCI (1199) AleI (1352) BspDI - ClaI (1798) BbsI - BpiI (2064) BbsI - BpiI (2090) PaeR7I - PspXI - XhoI (2113) EcoRI (2280) AgeI (2289) BstBI (2298) AfeI (2358) SV40 NLS SV40 NLS SV40 NLS PspOMI (4148) ApaI (4152) EcoRV (4646) PmlI (5582) pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro 11,553 bp
SspI  (10,966)
1 site
A A T A T T T T A T A A
PciI  (9269)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SfiI  (8847)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AanI  (8618)
2 sites
T T A T A A A A T A T T
AanI  (8562)
2 sites
T T A T A A A A T A T T
KpnI  (8227)
1 site
G G T A C C C C A T G G
Acc65I  (8223)
1 site
G G T A C C C C A T G G
HincII  (7628)
1 site
G T Y R A C C A R Y T G
SalI  (7626)
1 site
G T C G A C C A G C T G
SexAI  (7587)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BstEII  (7154)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII  (7136)
1 site
C G G W C C G G C C W