pCMV6-AC-FC

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

Sequence Author: OriGene

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DraIII (6404) XmnI (5851) AlwNI (4775) BstZ17I (3980) BsmI (3928) AanI (3867) BstBI (3690) AanI (6529) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI (1030) HindIII (1044) RsrII (1061) MluI (1067) BsiWI (1070) NotI (1076) PaeR7I - PspXI - XhoI (1082) PspOMI (1094) ApaI (1098) PmeI (1790) FseI (1800) SacII (1804) XL39 (reverse primer) (1896 .. 1915) BbvCI (2116) BlpI (2160) BsmBI (2180) BstXI (2267) AgeI (2378) PflMI (2381) XcmI (2473) SexAI * (2565) SfiI (2751) StuI (2797) AvrII (2798) BclI * (2849) BsaBI * (2867) KasI (3007) NarI (3008) SfoI (3009) PluTI (3011) PstI (3061) PflFI - Tth111I (3126) pCMV6-AC-FC 6601 bp
DraIII  (6404)
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  (5851)
1 site
G A A N N N N T T C C T T N N N N A A G
AlwNI  (4775)
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  (3980)
1 site
G T A T A C C A T A T G
BsmI  (3928)
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  (3867)
2 sites
T T A T A A A A T A T T
BstBI  (3690)
1 site
T T C G A A A A G C T T
AanI  (6529)
2 sites
T T A T A A A A T A T T
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
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
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
PspOMI  (1094)
1 site
G G G C C C C C C G G G
ApaI  (1098)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmeI  (1790)
1 site
G T T T A A A C C A A A T T T G
FseI  (1800)
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  (1804)
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  (2116)
1 site
C C T C A G C G G A G T C G
BlpI  (2160)
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  (2180)
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  (2267)
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  (2378)
1 site
A C C G G T T G G C C A
PflMI  (2381)
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  (2473)
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  (2565)
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  (2751)
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  (2797)
1 site
A G G C C T T C C G G A
AvrII  (2798)
1 site
C C T A G G G G A T C C
BclI  (2849)
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  (2867)
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  (3007)
1 site
G G C G C C C C G C G G
NarI  (3008)
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  (3009)
1 site
G G C G C C C C G C G G
PluTI  (3011)
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  (3061)
1 site
C T G C A G G A C G T C
PflFI  (3126)
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  (3126)
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.
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
1896 .. 1915  =  20 annealed bases
Tm  =  59°C
AmpR
5179 .. 6039  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5179 .. 5970  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5179 .. 6039  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5971 .. 6039  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5179 .. 6039  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2880 .. 3674  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2880 .. 3674  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
Fc tag
1088 .. 1783  =  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
1088 .. 1783  =  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
1833 .. 2455  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1833 .. 2455  =  623 bp
human growth hormone polyadenylation signal
ori
4420 .. 5008  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4420 .. 5008  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6171 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6171 .. 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
2484 .. 2813  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2484 .. 2813  =  330 bp
SV40 enhancer and early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter