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pCMV6-XL5-DDK-His

PrecisionShuttle™ mammalian transient expression vector for fusing an ORF to C-terminal FLAG® and 6xHis tags.

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pCMV6-XL5-DDK-His.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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PsiI (4543) DraIII (4418) XmnI (3865) ScaI (3746) AlwNI (2789) PciI (2373) BspQI - SapI (2257) PvuII (95) BsrGI (301) SpeI (357) NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI - BssHII (1030) NheI (1055) BmtI (1059) RsrII (1061) MluI (1067) BglII (1132) EcoRV (1146) XbaI (1162) SacII (1177) TspMI - XmaI (1203) SmaI (1205) AleI (1263) XL39 (reverse primer) (1269 .. 1288) BbsI (1386) EcoO109I (1393) BsgI (1447) BbvCI - Bpu10I (1489) BlpI (1533) BsmBI (1553) BstXI (1640) AgeI (1751) PflMI (1754) XcmI (1846) SexAI * (1938) SfiI (2124) StuI (2170) AvrII (2171) pCMV6-XL5-DDK-His 4626 bp
PsiI  (4543)
1 site
T T A T A A A A T A T T
DraIII  (4418)
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.
XmnI  (3865)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3746)
1 site
A G T A C T T C A T G A
AlwNI  (2789)
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  (2373)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2257)
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  (2257)
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.
PvuII  (95)
1 site
C A G C T G G T C G A C
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
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
BssHII  (1030)
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.
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
BglII  (1132)
1 site
A G A T C T T C T A G A
EcoRV  (1146)
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.
XbaI  (1162)
1 site
T C T A G A A G A T C T
SacII  (1177)
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.
TspMI  (1203)
1 site
C C C G G G G G G C C C
XmaI  (1203)
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.
SmaI  (1205)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
AleI  (1263)
1 site
C A C N N N N G T G G T G N N N N C A C
BbsI  (1386)
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.
EcoO109I  (1393)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BsgI  (1447)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N