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Plasmid Files

pCMV6-AN-mGFP

PrecisionShuttle™ mammalian expression vector for fusing an ORF to an N-terminal TagGFP2 (mTagGFP) tag.

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pCMV6-AN-mGFP Sequence and MappCMV6-AN-mGFP.dna
Map and Sequence File   
Sequence Author:  OriGene
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 AanI (6559) ScaI (5762) PciI (4389) BstZ17I (4010) BsmI (3958) AanI (3897) BstBI (3720) PflFI - Tth111I (3156) NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) BmgBI (1082) BstEII (1199) PasI (1580) BspEI (1734) AsiSI - SgfI (1750) MreI - SgrAI (1752) AscI (1756) HindIII (1770) RsrII (1787) MluI (1793) NotI (1803) PaeR7I - PspXI - XhoI (1811) PmeI (1820) FseI (1830) XL39 (reverse primer) (1926 .. 1945) BbsI (2043) EcoO109I (2050) BbvCI (2146) BlpI (2190) BsmBI (2210) BstXI (2297) AgeI (2408) SexAI * (2595) SfiI (2781) StuI (2827) AvrII (2828) BclI * (2879) KasI (3037) NarI (3038) SfoI (3039) PluTI (3041) PstI (3091) pCMV6-AN-mGFP 6631 bp
AanI  (6559)
2 sites
T T A T A A A A T A T T
ScaI  (5762)
1 site
A G T A C T T C A T G A
PciI  (4389)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (4010)
1 site
G T A T A C C A T A T G
BsmI  (3958)
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  (3897)
2 sites
T T A T A A A A T A T T
BstBI  (3720)
1 site
T T C G A A A A G C T T
PflFI  (3156)
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  (3156)
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
BmgBI  (1082)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BstEII  (1199)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
PasI  (1580)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
BspEI  (1734)
1 site
T C C G G A A G G C C T
AsiSI  (1750)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1750)
1 site
G C G A T C G C C G C T A G C G
MreI  (1752)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1752)
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  (1756)
1 site
G G C G C G C C C C G C G C G G
HindIII  (1770)
1 site
A A G C T T T T C G A A
RsrII  (1787)
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  (1793)
1 site
A C G C G T T G C G C A
NotI  (1803)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1811)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1811)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1811)
1 site
C T C G A G G A G C T C
PmeI  (1820)
1 site
G T T T A A A C C A A A T T T G
FseI  (1830)
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.
BbsI  (2043)
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  (2050)
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.
BbvCI  (2146)
1 site
C C T C A G C G G A G T C G
BlpI  (2190)
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  (2210)
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  (2297)
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  (2408)
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.
SexAI  (2595)
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  (2781)
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  (2827)
1 site
A G G C C T T C C G G A
AvrII  (2828)
1 site
C C T A G G G G A T C C
BclI  (2879)
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.
KasI  (3037)
1 site
G G C G C C C C G C G G
NarI  (3038)
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  (3039)
1 site
G G C G C C C C G C G G
PluTI  (3041)
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  (3091)
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
1926 .. 1945  =  20 annealed bases
Tm  =  59°C
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5209 .. 6000  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6001 .. 6069  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
2910 .. 3704  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
2910 .. 3704  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
TagGFP2
1020 .. 1733  =  714 bp
238 amino acids  =  26.9 kDa
Product: monomeric green fluorescent protein, also
known as mTagGFP
mammalian codon-optimized
TagGFP2
1020 .. 1733  =  714 bp
238 amino acids  =  26.9 kDa
Product: monomeric green fluorescent protein, also
known as mTagGFP
mammalian codon-optimized
hGH poly(A) signal
1863 .. 2485  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1863 .. 2485  =  623 bp
human growth hormone polyadenylation signal
ori
4450 .. 5038  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4450 .. 5038  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
6201 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6201 .. 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
2514 .. 2843  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2514 .. 2843  =  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
3878 .. 3999  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3878 .. 3999  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6070 .. 6174  =  105 bp
AmpR promoter
6070 .. 6174  =  105 bp
MCS
1734 .. 1836  =  103 bp
multiple cloning site
MCS
1734 .. 1836  =  103 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
2694 .. 2829  =  136 bp
SV40 origin of replication
SV40 ori
2694 .. 2829  =  136 bp
SV40 origin of replication
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