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pCLIP-Cas9-Nuclease-TRE3G-Puro

Lentiviral CRISPR vector encoding Cas9 and a puromycin resistance marker expressed from a Tet-responsive promoter.

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pCLIP-Cas9-Nuclease-TRE3G-Puro.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (10,676) PvuI (10,242) FspI (10,094) DrdI (9090) AflIII - PciI (8982) SmaI (8962) TspMI - XmaI (8960) AvrII (8939) PmeI (8614) PsiI (8564) KpnI (8229) Acc65I (8225) Bsu36I (8221) SalI (8101) EcoRI (7479) BstEII (7013) RsrII (6995) BsiWI (6935) PflFI - Tth111I (6921) BamHI (6816) BsaBI * (6138) PmlI (5429) SpeI (41) NdeI (276) SnaBI (382) NheI (405) BmtI (409) NruI * (867) BspQI - SapI (1136) FseI (1184) MfeI (1223) BbvCI (1458) AleI (1611) AbsI - PspXI (2238) XbaI (2623) AfeI (2630) PspOMI (3995) ApaI (3999) pCLIP-Cas9-Nuclease-TRE3G-Puro 10,794 bp
SspI  (10,676)
1 site
A A T A T T T T A T A A
PvuI  (10,242)
1 site
C G A T C G G C T A G C
FspI  (10,094)
1 site
T G C G C A A C G C G T
DrdI  (9090)
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  (8982)
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  (8982)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (8962)
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  (8960)
1 site
C C C G G G G G G C C C
XmaI  (8960)
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  (8939)
1 site
C C T A G G G G A T C C
PmeI  (8614)
1 site
G T T T A A A C C A A A T T T G
PsiI  (8564)
1 site
T T A T A A A A T A T T
KpnI  (8229)
1 site
G G T A C C C C A T G G
Acc65I  (8225)
1 site
G G T A C C C C A T G G
Bsu36I  (8221)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SalI  (8101)
1 site
G T C G A C C A G C T G
EcoRI  (7479)
1 site
G A A T T C C T T A A G
BstEII  (7013)
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  (6995)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsiWI  (6935)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PflFI  (6921)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (6921)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BamHI  (6816)
1 site