pCMV6-AC-3HA

PrecisionShuttle™ mammalian expression vector for fusing an ORF to a C-terminal 3xHA tag.

Sequence Author: OriGene

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BsrGI (301) DraIII (5792) XmnI (5239) ScaI (5120) AlwNI (4166) PciI (3750) BstZ17I (3371) BsmI (3319) BstBI (3081) PflFI - Tth111I (2517) SpeI (357) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) T7 promoter EcoRI (979) Acc65I (998) KpnI (1002) AsiSI (1024) MreI - SgrAI (1026) AscI (1030) HindIII (1044) NheI (1055) BmtI (1059) RsrII (1061) MluI (1067) BsiWI (1070) NotI (1076) PaeR7I - PspXI - XhoI (1082) 3xHA PmeI (1181) FseI (1191) SacII (1195) AleI (1281) XL39 (reverse primer) (1287 .. 1306) BbsI (1404) EcoO109I (1411) BsgI (1465) BbvCI - Bpu10I (1507) BlpI (1551) BsmBI - Esp3I (1571) BstXI (1658) AgeI (1769) PflMI (1772) XcmI (1864) CsiI - SexAI * (1956) SfiI (2142) StuI (2188) AvrII (2189) BclI * (2240) BsaBI * (2258) KasI (2398) NarI (2399) SfoI (2400) PluTI (2402) PstI (2452) pCMV6-AC-3HA 5989 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.
DraIII  (5792)
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  (5239)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5120)
1 site
A G T A C T T C A T G A
AlwNI  (4166)
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  (3750)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3371)
1 site
G T A T A C C A T A T G
BsmI  (3319)
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.
BstBI  (3081)
1 site
T T C G A A A A G C T T
PflFI  (2517)
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  (2517)
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
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
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
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  (1181)
1 site
G T T T A A A C C A A A T T T G
FseI  (1191)
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  (1195)
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  (1281)
1 site
C A C N N N N G T G G T G N N N N C A C
BbsI  (1404)
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  (1411)
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  (1465)
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  (1507)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1507)
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.
BlpI  (1551)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsmBI  (1571)
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.
Esp3I  (1571)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BstXI  (1658)
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  (1769)
1 site
A C C G G T T G G C C A
PflMI  (1772)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
XcmI  (1864)
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.
CsiI  (1956)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (1956)
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  (2142)
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  (2188)
1 site
A G G C C T T C C G G A
AvrII  (2189)
1 site
C C T A G G G G A T C C
BclI  (2240)
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.
BsaBI  (2258)
1 site
G A T N N N N A T C C T A N N N