pCMV6-AN-FC

PrecisionShuttle™ mammalian expression vector for fusing an ORF to an N-terminal murine Fc tag.

Sequence Author: OriGene

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AanI (6532) DraIII (6407) XmnI (5854) AlwNI (4778) BstZ17I (3983) BsmI (3931) AanI (3870) BstBI (3693) PflFI - Tth111I (3129) NdeI (592) SnaBI (698) CMV promoter VP1.5 (forward primer) (839 .. 856) T7 promoter EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) PspOMI (1029) ApaI (1033) AsiSI - SgfI (1723) MreI - SgrAI (1725) AscI (1729) HindIII (1743) RsrII (1760) MluI (1766) NotI (1776) PaeR7I - PspXI - XhoI (1784) PmeI (1793) FseI (1803) SacII (1807) XL39 (reverse primer) (1899 .. 1918) BbvCI (2119) BlpI (2163) BsmBI (2183) BstXI (2270) AgeI (2381) PflMI (2384) XcmI (2476) SexAI * (2568) SfiI (2754) StuI (2800) AvrII (2801) BclI * (2852) BsaBI * (2870) KasI (3010) NarI (3011) SfoI (3012) PluTI (3014) PstI (3064) pCMV6-AN-FC 6604 bp
AanI  (6532)
2 sites
T T A T A A A A T A T T
DraIII  (6407)
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  (5854)
1 site
G A A N N N N T T C C T T N N N N A A G
AlwNI  (4778)
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.
BstZ17I  (3983)
1 site
G T A T A C C A T A T G
BsmI  (3931)
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  (3870)
2 sites
T T A T A A A A T A T T
BstBI  (3693)
1 site
T T C G A A A A G C T T
PflFI  (3129)
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  (3129)
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
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
PspOMI  (1029)
1 site
G G G C C C C C C G G G
ApaI  (1033)
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
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
NotI  (1776)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1784)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1784)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1784)
1 site
C T C G A G G A G C T C
PmeI  (1793)
1 site
G T T T A A A C C A A A T T T G
FseI  (1803)
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  (1807)
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.
BbvCI  (2119)
1 site
C C T C A G C G G A G T C G
BlpI  (2163)
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  (2183)
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  (2270)
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  (2381)
1 site
A C C G G T T G G C C A
PflMI  (2384)
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  (2476)
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  (2568)
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  (2754)
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  (2800)
1 site
A G G C C T T C C G G A
AvrII  (2801)
1 site
C C T A G G G G A T C C
BclI  (2852)
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  (2870)
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  (3010)
1 site
G G C G C C C C G C G G
NarI  (3011)
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  (3012)
1 site
G G C G C C C C G C G G
PluTI  (3014)
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  (3064)
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
1899 .. 1918  =  20 annealed bases
Tm  =  59°C
AmpR
5182 .. 6042  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5182 .. 5973  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5182 .. 6042  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5974 .. 6042  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5182 .. 6042  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2883 .. 3677  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2883 .. 3677  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
Fc tag
1023 .. 1718  =  696 bp
232 amino acids  =  26.3 kDa
Product: CH2 and CH3 domains of the mouse IgG2a heavy chain plus the hinge region
Fc tag
1023 .. 1718  =  696 bp
232 amino acids  =  26.3 kDa
Product: CH2 and CH3 domains of the mouse IgG2a heavy chain plus the hinge region
hGH poly(A) signal
1836 .. 2458  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1836 .. 2458  =  623 bp
human growth hormone polyadenylation signal
ori
4423 .. 5011  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4423 .. 5011  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6174 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6174 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
2487 .. 2816  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2487 .. 2816  =  330 bp
SV40 enhancer and early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
3851 .. 3972  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3851 .. 3972  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6043 .. 6147  =  105 bp
AmpR promoter
6043 .. 6147  =  105 bp
MCS
1719 .. 1809  =  91 bp
multiple cloning site
MCS
1719 .. 1809  =  91 bp
multiple cloning site
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  3055 .. 3441  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  5312 .. 5578  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  882 .. 1754  =  873 bp
ORF:  290 amino acids  =  32.9 kDa
ORF:  2883 .. 3677  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1262 .. 1507  =  246 bp
ORF:  81 amino acids  =  8.7 kDa
ORF:  2200 .. 2424  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  5182 .. 6042  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  930 .. 1253  =  324 bp
ORF:  107 amino acids  =  12.4 kDa
ORF:  3192 .. 3728  =  537 bp
ORF:  178 amino acids  =  19.7 kDa
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