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pCLIP-Cas9-Nuclease-TRE3G-ZsGreen

Lentiviral CRISPR vector encoding Cas9 and the green fluorescent protein ZsGreen expressed from a Tet-responsive promoter.

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pCLIP-Cas9-Nuclease-TRE3G-ZsGreen.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (10,775) PvuI (10,341) FspI (10,193) DrdI (9189) AflIII - PciI (9081) SmaI (9061) TspMI - XmaI (9059) AvrII (9038) SexAI * (8805) PmeI (8713) PsiI (8663) KpnI (8328) Acc65I (8324) Bsu36I (8320) SacII (8105) EcoRI (7578) BsrGI (7432) PflFI - Tth111I (7116) SgrAI (6969) BamHI (6816) BsaBI * (6138) PmlI (5429) SpeI (41) NdeI (276) SnaBI (382) NheI (405) BmtI (409) BssHII (745) NruI * (867) BspQI - SapI (1136) FseI (1184) MfeI (1223) BbvCI (1458) AleI (1611) AbsI - PspXI (2238) XbaI (2623) AfeI (2630) PspOMI (3995) ApaI (3999) PasI (4409) DraIII (5039) pCLIP-Cas9-Nuclease-TRE3G-ZsGreen 10,893 bp
SspI  (10,775)
1 site
A A T A T T T T A T A A
PvuI  (10,341)
1 site
C G A T C G G C T A G C
FspI  (10,193)
1 site
T G C G C A A C G C G T
DrdI  (9189)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AflIII  (9081)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (9081)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (9061)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (9059)
1 site
C C C G G G G G G C C C
XmaI  (9059)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
AvrII  (9038)
1 site
C C T A G G G G A T C C
SexAI  (8805)
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.
PmeI  (8713)
1 site
G T T T A A A C C