Resources
Plasmid Files

pCMV6-AC-IRES-GFP-Puro

PrecisionShuttle™ mammalian expression vector with TurboGFP and puromycin resistance markers, for fusing an ORF to C-terminal c-Myc and FLAG® tags.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pCMV6-AC-IRES-GFP-Puro Sequence and MappCMV6-AC-IRES-GFP-Puro.dna
Map and Sequence File   
Sequence Author:  OriGene
Download Free Trial Get SnapGene Viewer

 AanI (7000) ScaI (6203) AlwNI (5246) BstZ17I (4451) AanI (4338) BstBI (4161) SacII (3755) BstEII (3675) PflFI - Tth111I (3583) BsrGI (301) SpeI (357) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI (1030) NheI (1055) BmtI (1059) RsrII (1061) MluI (1067) PaeR7I - PspXI - XhoI (1082) EcoRV (1126) PmeI (1160) BmgBI (1727) FseI (2114) BsaBI * (2234) AleI (2560) XL39 (reverse primer) (2566 .. 2585) BbvCI - Bpu10I (2786) AgeI (3048) XcmI (3143) SfiI (3421) BclI * (3519) pCMV6-AC-IRES-GFP-Puro 7072 bp
AanI  (7000)
2 sites
T T A T A A A A T A T T
ScaI  (6203)
1 site
A G T A C T T C A T G A
AlwNI  (5246)
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  (4451)
1 site
G T A T A C C A T A T G
AanI  (4338)
2 sites
T T A T A A A A T A T T
BstBI  (4161)
1 site
T T C G A A A A G C T T
SacII  (3755)
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.
BstEII  (3675)
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.
PflFI  (3583)
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  (3583)
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.
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.
SpeI  (357)
1 site
A C T A G T T G A T C A
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.
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
NheI  (1055)
1 site
G C T A G C C G A T C G
BmtI  (1059)
1 site
G C T A G C C G A T C G
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
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
EcoRV  (1126)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
PmeI  (1160)
1 site
G T T T A A A C C A A A T T T G
BmgBI  (1727)
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.
FseI  (2114)
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.
BsaBI  (2234)
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.
AleI  (2560)
1 site
C A C N N N N G T G G T G N N N N C A C
BbvCI  (2786)
1 site
C C T C A G C G G A G T C G
Bpu10I  (2786)
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.
AgeI  (3048)
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  (3143)
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.
SfiI  (3421)
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.
BclI  (3519)
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.
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
2566 .. 2585  =  20 annealed bases
Tm  =  59°C
AmpR
5650 .. 6510  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5650 .. 6441  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5650 .. 6510  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6442 .. 6510  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5650 .. 6510  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TurboGFP
1771 .. 2469  =  699 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
1771 .. 2469  =  699 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
2503 .. 3125  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
2503 .. 3125  =  623 bp
human growth hormone polyadenylation signal
PuroR
3541 .. 4140  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3541 .. 4140  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4891 .. 5479  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4891 .. 5479  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES2
1184 .. 1770  =  587 bp
   Segment 1:  
   1184 .. 1758  =  575 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1184 .. 1770  =  587 bp
   Segment 2:  ATG  
   1759 .. 1761  =  3 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1184 .. 1770  =  587 bp
   Segment 3:  
   1762 .. 1770  =  9 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1184 .. 1770  =  587 bp
3 segments
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
f1 ori
6642 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6642 .. 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
3154 .. 3483  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3154 .. 3483  =  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
4319 .. 4440  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4319 .. 4440  =  122 bp
SV40 polyadenylation signal
MCS
979 .. 1087  =  109 bp
multiple cloning site
MCS
979 .. 1087  =  109 bp
multiple cloning site
AmpR promoter
6511 .. 6615  =  105 bp
AmpR promoter
6511 .. 6615  =  105 bp
FLAG
1133 .. 1156  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
FLAG
1133 .. 1156  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage 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
3334 .. 3469  =  136 bp
SV40 origin of replication
SV40 ori
3334 .. 3469  =  136 bp
SV40 origin of replication
Myc
1085 .. 1114  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
1085 .. 1114  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
ATG
1759 .. 1761  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1759 .. 1761  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter