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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AanI (6544) DraIII (6419) ScaI (5747) AlwNI (4793) PciI (4377) BstZ17I (3998) BsmI (3946) AanI (3885) PfoI (3801) BstBI (3708) BsrGI (301) SpeI (357) NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) 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) PflFI - Tth111I (3144) pCMV6-AN-GFP-AC-His 6616 bp
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
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.
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