pCMV6-AN-His

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

Sequence Author: OriGene

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BsrGI (301) AanI (5851) DraIII (5726) XmnI (5173) ScaI (5054) AlwNI (4100) PciI (3684) BstZ17I (3305) BsmI (3253) AanI (3192) PfoI (3108) BstBI (3015) PflFI - Tth111I (2451) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) T7 promoter EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) ATG 6xHis AsiSI - SgfI (1045) MreI - SgrAI (1047) AscI (1051) HindIII (1065) RsrII (1082) MluI (1088) NotI (1098) PaeR7I - PspXI - XhoI (1106) PmeI (1115) FseI (1125) SacII (1129) AleI (1215) XL39 (reverse primer) (1221 .. 1240) BbsI (1338) EcoO109I (1345) BsgI (1399) BbvCI - Bpu10I (1441) BlpI (1485) BsmBI (1505) BstXI (1592) AgeI (1703) PflMI (1706) XcmI (1798) SexAI * (1890) SfiI (2076) StuI (2122) AvrII (2123) BclI * (2174) BsaBI * (2192) KasI (2332) NarI (2333) SfoI (2334) PluTI (2336) PstI (2386) pCMV6-AN-His 5923 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  (5851)
2 sites
T T A T A A A A T A T T
DraIII  (5726)
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  (5173)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5054)
1 site
A G T A C T T C A T G A
AlwNI  (4100)
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  (3684)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3305)
1 site
G T A T A C C A T A T G
BsmI  (3253)
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  (3192)
2 sites
T T A T A A A A T A T T
PfoI  (3108)
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  (3015)
1 site
T T C G A A A A G C T T
PflFI  (2451)
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  (2451)
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.
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  (1045)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1045)
1 site
G C G A T C G C C G C T A G C G
MreI  (1047)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1047)
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  (1051)
1 site
G G C G C G C C C C G C G C G G
HindIII  (1065)
1 site
A A G C T T T T C G A A
RsrII  (1082)
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  (1088)
1 site
A C G C G T T G C G C A
NotI  (1098)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1106)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1106)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1106)
1 site
C T C G A G G A G C T C
PmeI  (1115)
1 site
G T T T A A A C C A A A T T T G
FseI  (1125)
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  (1129)
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  (1215)
1 site
C A C N N N N G T G G T G N N N N C A C
BbsI  (1338)
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  (1345)
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  (1399)
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  (1441)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1441)
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  (1485)
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  (1505)
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  (1592)
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  (1703)
1 site
A C C G G T T G G C C A
PflMI  (1706)
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  (1798)
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  (1890)
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  (2076)
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  (2122)
1 site
A G G C C T T C C G G A
AvrII  (2123)
1 site
C C T A G G G G A T C C
BclI  (2174)
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  (2192)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
KasI  (2332)
1 site
G G C G C C C C G C G G
NarI  (2333)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2334)
1 site
G G C G C C C C G C G G
PluTI  (2336)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
PstI  (2386)
1 site
C T G C A G G A C G T 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
1221 .. 1240  =  20 annealed bases
Tm  =  59°C
AmpR
4501 .. 5361  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4501 .. 5292  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4501 .. 5361  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5293 .. 5361  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4501 .. 5361  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2205 .. 2999  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2205 .. 2999  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
hGH poly(A) signal
1158 .. 1780  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1158 .. 1780  =  623 bp
human growth hormone polyadenylation signal
ori
3745 .. 4330  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3745 .. 4330  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5493 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5493 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis