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pCLIP-Cas9-Nuclease-TRE3G-Blast

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

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pCLIP-Cas9-Nuclease-TRE3G-Blast.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (10,478) FspI (9896) AflIII - PciI (8784) SmaI (8764) TspMI - XmaI (8762) AvrII (8741) SexAI * (8508) PmeI (8416) PsiI (8366) KpnI (8031) Acc65I (8027) Bsu36I (8023) SalI (7903) SacII (7808) EcoRI (7281) BamHI (6816) PmlI (5429) SpeI (41) NdeI (276) SnaBI (382) NheI (405) BmtI (409) BssHII (745) 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-Blast 10,596 bp
SspI  (10,478)
1 site
A A T A T T T T A T A A
FspI  (9896)
1 site
T G C G C A A C G C G T
AflIII  (8784)
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  (8784)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (8764)
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  (8762)
1 site
C C C G G G G G G C C C
XmaI  (8762)
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  (8741)
1 site
C C T A G G G G A T C C
SexAI  (8508)
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  (8416)
1 site
G T T T A A A C C A A A T T T G
PsiI  (8366)
1 site
T T A T A A A A T A T T
KpnI  (8031)
1 site
G G T A C C C C A T G G
Acc65I  (8027)
1 site
G G T A C C C C A T G G
Bsu36I  (8023)
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  (7903)
1 site
G T C G A C C A G C T G
SacII  (7808)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
EcoRI  (7281)
1 site
G A A T T C C T T A A G
BamHI  (6816)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
PmlI  (5429)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
SpeI  (41)
1 site
A C T A G T T G A T C A
NdeI  (276)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SnaBI  (382)
1 site
T A C G T A A T G C A T
NheI  (405)
1 site
G C T A G C C G A T C G
BmtI  (409)
1 site
G C T A G C C G A T C G
BssHII  (745)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BspQI  (1136)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1136)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
FseI  (1184)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
MfeI  (1223)
1 site
C A A T T G G T T A A C
BbvCI  (1458)
1 site
C C T C A G C G G A G T C G
AleI  (1611)
1 site
C A C N N N N G T G G T G N N N N C A C
AbsI  (2238)
1 site
C C T C G A G G G G A G C T C C
PspXI  (2238)
1 site
V C T C G A G B B G A G C T C V
XbaI  (2623)
1 site
T C T A G A A G A T C T
AfeI  (2630)
1 site
A