pCMV6-AC-HA

PrecisionShuttle™ mammalian expression vector for fusing an ORF to a C-terminal HA tag.

Sequence Author: OriGene

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SpeI (357) BsrGI (301) AanI (5854) DraIII (5729) XmnI (5176) ScaI (5057) AlwNI (4103) PciI (3687) BstZ17I (3308) BsmI (3256) AanI (3195) PfoI (3111) BstBI (3018) PflFI - Tth111I (2454) PstI (2389) NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) T7 promoter EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI (1030) HindIII (1044) NheI (1055) BmtI (1059) RsrII (1061) MluI (1067) BsiWI (1070) NotI (1076) PaeR7I - PspXI - XhoI (1082) HA PmeI (1118) FseI (1128) SacII (1132) AleI (1218) XL39 (reverse primer) (1224 .. 1243) BbsI (1341) EcoO109I (1348) BsgI (1402) BbvCI - Bpu10I (1444) BlpI (1488) BsmBI (1508) BstXI (1595) AgeI (1706) PflMI (1709) XcmI (1801) SexAI * (1893) SfiI (2079) StuI (2125) AvrII (2126) BclI * (2177) BsaBI * (2195) KasI (2335) NarI (2336) SfoI (2337) PluTI (2339) pCMV6-AC-HA 5926 bp
SpeI  (357)
1 site
A C T A G T T G A T C A
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  (5854)
2 sites
T T A T A A A A T A T T
DraIII  (5729)
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  (5176)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5057)
1 site
A G T A C T T C A T G A
AlwNI  (4103)
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  (3687)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3308)
1 site
G T A T A C C A T A T G
BsmI  (3256)
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  (3195)
2 sites
T T A T A A A A T A T T
PfoI  (3111)
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  (3018)
1 site
T T C G A A A A G C T T
PflFI  (2454)
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  (2454)
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.
PstI  (2389)
1 site
C T G C A G G A C G T C
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
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
HindIII  (1044)
1 site
A A G C T T T T C G A A
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
BsiWI  (1070)
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  (1076)
1 site
G C G G C C G C C G C C G G C G
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
PmeI  (1118)
1 site
G T T T A A A C C A A A T T T G
FseI  (1128)
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.
SacII  (1132)
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  (1218)
1 site
C A C N N N N G T G G T G N N N N C A C
BbsI  (1341)
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  (1348)
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  (1402)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BbvCI  (1444)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1444)
1 site
C C T N A G C G G A N