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Plasmid Files

pCMV6-AN-GFP

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

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pCMV6-AN-GFP Sequence and MappCMV6-AN-GFP.dna
Map and Sequence File   
Sequence Author:  OriGene
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 DraIII (6404) ScaI (5732) AlwNI (4778) PciI (4362) BstZ17I (3983) BsmI (3931) AanI (3870) PfoI (3786) BstBI (3693) AanI (6529) BsrGI (301) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) AarI (1471) PspOMI (1604) ApaI (1608) AsiSI - SgfI (1723) MreI - SgrAI (1725) AscI (1729) HindIII (1743) RsrII (1760) MluI (1766) NotI (1776) PaeR7I - PspXI - XhoI (1784) PmeI (1793) SacII (1807) AleI (1893) XL39 (reverse primer) (1899 .. 1918) BbsI (2016) BbvCI - Bpu10I (2119) BsmBI (2183) BstXI (2270) AgeI (2381) XcmI (2476) SexAI * (2568) SfiI (2754) StuI (2800) AvrII (2801) BclI * (2852) PflFI - Tth111I (3129) pCMV6-AN-GFP 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.
ScaI  (5732)
1 site
A G T A C T T C A T G A
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.
PciI  (4362)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
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
PfoI  (3786)
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  (3693)
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
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.
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
AarI  (1471)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
PspOMI  (1604)
1 site
G G G C C C C C C G G G
ApaI  (1608)
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
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.
AleI  (1893)
1 site
C A C N N N N G T G G T G N N N N C A C
BbsI  (2016)
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.
BbvCI  (2119)
1 site
C C T C A G C G G A G T C G
Bpu10I  (2119)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
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.
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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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.
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.
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
5179 .. 6039  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
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®)
TurboGFP
1020 .. 1715  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
1020 .. 1715  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
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 .. 5008  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4423 .. 5008  =  586 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
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
6040 .. 6144  =  105 bp
AmpR promoter
6040 .. 6144  =  105 bp
MCS
1719 .. 1797  =  79 bp
multiple cloning site
MCS
1719 .. 1797  =  79 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
SV40 ori
2667 .. 2802  =  136 bp
SV40 origin of replication
SV40 ori
2667 .. 2802  =  136 bp
SV40 origin of replication
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