pCMV6-AN-GFP-AC-His

PrecisionShuttle™ mammalian expression vector for fusing an ORF to an N-terminal TurboGFP tag and a C-terminal 6xHis tag.

Sequence Author: OriGene

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HomePlasmidsMammalian Expression VectorspCMV6-AN-GFP-AC-His
BsrGI (301) AanI (6544) DraIII (6419) ScaI (5747) AlwNI (4793) PciI (4377) BstZ17I (3998) BsmI (3946) AanI (3885) PfoI (3801) BstBI (3708) PflFI - Tth111I (3144) SpeI (357) NdeI (592) SnaBI (698) CMV promoter VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) T7 promoter EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) AarI (1471) PspOMI (1604) ApaI (1608) AsiSI - SgfI (1723) MreI - SgrAI (1725) AscI (1729) HindIII (1743) NheI (1754) BmtI (1758) RsrII (1760) MluI (1766) BsiWI (1769) NotI (1775) PaeR7I - PspXI - XhoI (1781) 6xHis PmeI (1808) SacII (1822) AleI (1908) XL39 (reverse primer) (1914 .. 1933) BbsI (2031) BbvCI - Bpu10I (2134) BsmBI (2198) BstXI (2285) AgeI (2396) XcmI (2491) SexAI * (2583) SfiI (2769) StuI (2815) AvrII (2816) BclI * (2867) pCMV6-AN-GFP-AC-His 6616 bp
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.
AanI  (6544)
2 sites		 T T A T A A A A T A T T
DraIII  (6419)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
ScaI  (5747)
1 site		 A G T A C T T C A T G A
AlwNI  (4793)
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.
PciI  (4377)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BstZ17I  (3998)
1 site		 G T A T A C C A T A T G
BsmI  (3946)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
AanI  (3885)
2 sites		 T T A T A A A A T A T T
PfoI  (3801)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
BstBI  (3708)
1 site		 T T C G A A A A G C T T
PflFI  (3144)
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  (3144)
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.
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.
Acc65I  (998)
1 site		 G G T A C C C C A T G G
KpnI  (1002)
1 site		 G G T A C C C C A T G G
AarI  (1471)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
PspOMI  (1604)
1 site		 G G G C C C C C C G G G
ApaI  (1608)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
AsiSI  (1723)
1 site		 G C G A T C G C C G C T A G C G
SgfI  (1723)
1 site		 G C G A T C G C C G C T A G C G
MreI  (1725)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (1725)
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  (1729)
1 site		 G G C G C G C C C C G C G C G G
HindIII  (1743)
1 site		 A A G C T T T T C G A A
NheI  (1754)
1 site		 G C T A G C C G A T C G
BmtI  (1758)
1 site		 G C T A G C C G A T C G
RsrII  (1760)
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  (1766)
1 site		 A C G C G T T G C G C A
BsiWI  (1769)
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.
NotI  (1775)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (1781)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1781)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1781)
1 site		 C T C G A G G A G C T C
PmeI  (1808)
1 site		 G T T T A A A C C A A A T T T G
SacII  (1822)
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.
AleI  (1908)
1 site		 C A C N N N N G T G G T G N N N N C A C
BbsI  (2031)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbvCI  (2134)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (2134)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsmBI  (2198)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
BstXI  (2285)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
AgeI  (2396)
1 site		 A C C G G T T G G C C A
XcmI  (2491)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SexAI  (2583)
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.
SfiI  (2769)
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.
StuI  (2815)
1 site		 A G G C C T T C C G G A
AvrII  (2816)
1 site		 C C T A G G G G A T C C
BclI  (2867)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
VP1.5 (forward primer)
18-mer  /  44% GC
1 binding site
839 .. 856  =  18 annealed bases
Tm  =  51°C
XL39 (reverse primer)
20-mer  /  55% GC
1 binding site
1914 .. 1933  =  20 annealed bases
Tm  =  59°C
AmpR
5194 .. 6054  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5194 .. 5985  =  792 bp