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Plasmid Files

pcDNA™6.2/EmGFP-Bsd/V5-DEST

Vivid Colors™ Gateway® destination vector for expressing proteins in mammalian cells, with an EmGFP-Bsd fusion as a transfection marker.

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pcDNA6.2 EmGFP-Bsd V5-DEST Sequence and MappcDNA6.2 EmGFP-Bsd V5-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SgrDI (8196) PciI (6382) BspQI - SapI (6266) EagI (5431) BbsI (5400) StuI (4316) SphI (4061) MfeI (161) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) PstI (1434) BfuAI - BspMI (1437) BstXI (1572) TspMI - XmaI (1683) SmaI - SrfI (1685) BbvCI (1828) BssHII (2058) BamHI (2099) EcoRI (2555) PspOMI (3188) ApaI (3192) SacII (3195) BstBI (3199) PmeI (3267) BstAPI (3533) PaeR7I - PspXI - XhoI (4011) pcDNA™6.2/EmGFP-Bsd/V5-DEST 8198 bp
SgrDI  (8196)
1 site
C G T C G A C G G C A G C T G C
PciI  (6382)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (6266)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (6266)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
EagI  (5431)
1 site
C G G C C G G C C G G C
BbsI  (5400)
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.
StuI  (4316)
1 site
A G G C C T T C C G G A
SphI  (4061)
1 site
G C A T G C C G T A C G
MfeI  (161)
1 site
C A A T T G G T T A A C
NdeI  (484)
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  (590)
1 site
T A C G T A A T G C A T
Eco53kI  (816)
1 site
G A G C T C C T C G A G
SacI  (818)
1 site
G A G C T C C T C G A G
PstI  (1434)
1 site
C T G C A G G A C G T C
BfuAI  (1437)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1437)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BstXI  (1572)
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.
TspMI  (1683)
1 site
C C C G G G G G G C C C
XmaI  (1683)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1685)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (1685)
1 site
G C C C G G G C C G G G C C C G
BbvCI  (1828)
1 site
C C T C A G C G G A G T C G
BssHII  (2058)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BamHI  (2099)
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.
EcoRI  (2555)
1 site
G A A T T C C T T A A G
PspOMI  (3188)
1 site
G G G C C C C C C G G G
ApaI  (3192)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SacII  (3195)
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.
BstBI  (3199)
1 site
T T C G A A A A G C T T
PmeI  (3267)
1 site
G T T T A A A C C A A A T T T G
BstAPI  (3533)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PaeR7I  (4011)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4011)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4011)
1 site
C T C G A G G A G C T C
AmpR
7202 .. 8062  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7202 .. 7993  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7202 .. 8062  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7994 .. 8062  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7202 .. 8062  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
EmGFP
4601 .. 5317  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 1:  
   4601 .. 4603  =  3 bp
   1 amino acid  =  149.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
4601 .. 5317  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 2:  1a  
   4604 .. 4606  =  3 bp
   1 amino acid  =  117.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
4601 .. 5317  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 3:  
   4607 .. 5317  =  711 bp
   237 amino acids  =  26.7 kDa
Product: Emerald GFP
mammalian codon-optimized
EmGFP
4601 .. 5317  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: Emerald GFP
mammalian codon-optimized
CmR
2114 .. 2773  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2114 .. 2773  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
6443 .. 7031  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6443 .. 7031  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
4022 .. 4521  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
4022 .. 4521  =  500 bp
mouse phosphoglycerate kinase 1 promoter
f1 ori
3579 .. 4007  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3579 .. 4007  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
5318 .. 5713  =  396 bp
131 amino acids  =  13.5 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
5318 .. 5713  =  396 bp
131 amino acids  =  13.5 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
ccdB
1464 .. 1769  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
1464 .. 1769  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3054 .. 3178  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3054 .. 3178  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5871 .. 5992  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5871 .. 5992  =  122 bp
SV40 polyadenylation signal
AmpR promoter
8063 .. 8167  =  105 bp
AmpR promoter
8063 .. 8167  =  105 bp
cat promoter
2774 .. 2876  =  103 bp
promoter of the E. coli cat gene
cat promoter
2774 .. 2876  =  103 bp
promoter of the E. coli cat gene
HSV TK poly(A) signal
3329 .. 3377  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3329 .. 3377  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
4535 .. 4582  =  48 bp
synthetic bacterial promoter
EM7 promoter
4535 .. 4582  =  48 bp
synthetic bacterial promoter
V5 tag
3204 .. 3245  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3204 .. 3245  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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