pCMV6-AC-IRES-GFP-Puro

PrecisionShuttle™ mammalian expression vector with TurboGFP and puromycin resistance markers, for fusing an ORF to C-terminal c-Myc and FLAG® tags.

Sequence Author: OriGene

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AanI (7000) ScaI (6203) AlwNI (5246) BstZ17I (4451) AanI (4338) BstBI (4161) SacII (3755) BstEII (3675) PflFI - Tth111I (3583) BsrGI (301) SpeI (357) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI (1030) NheI (1055) BmtI (1059) RsrII (1061) MluI (1067) PaeR7I - PspXI - XhoI (1082) EcoRV (1126) PmeI (1160) BmgBI (1727) FseI (2114) BsaBI * (2234) AleI (2560) XL39 (reverse primer) (2566 .. 2585) BbvCI - Bpu10I (2786) AgeI (3048) XcmI (3143) SfiI (3421) BclI * (3519) pCMV6-AC-IRES-GFP-Puro 7072 bp
AanI  (7000)
2 sites
T T A T A A A A T A T T
ScaI  (6203)
1 site
A G T A C T T C A T G A
AlwNI  (5246)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BstZ17I  (4451)
1 site
G T A T A C C A T A T G
AanI  (4338)
2 sites
T T A T A A A A T A T T
BstBI  (4161)
1 site
T T C G A A A A G C T T
SacII  (3755)
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.
BstEII  (3675)
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.
PflFI  (3583)
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  (3583)
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.
BsrGI  (301)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (357)
1 site
A C T A G T T G A T C A
NdeI  (592)
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  (698)
1 site
T A C G T A A T G C A T
Eco53kI  (924)
1 site
G A G C T C C T C G A G
SacI  (926)
1 site
G A G C T C C T C G A G
EcoRI  (979)
1 site
G A A T T C C T T A A G
BamHI  (992)
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.
AsiSI  (1024)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1024)
1 site
G C G A T C G C C G C T A G C G
MreI  (1026)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1026)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
AscI  (1030)
1 site
G G C G C G C C C C G C G C G G
NheI  (1055)
1 site
G C T A G C C G A T C G
BmtI  (1059)
1 site
G C T A G C C G A T C G
RsrII  (1061)
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.
MluI  (1067)
1 site
A C G C G T T G C G C A
PaeR7I  (1082)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1082)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1082)
1 site
C T C G A G G A G C T C
EcoRV  (1126)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
PmeI  (1160)
1 site
G T T T A A A C C A A A T T T G
BmgBI  (1727)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
FseI  (2114)
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.
BsaBI  (2234)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
AleI  (2560)
1 site
C A C N N N N G T G G T G N N