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Plasmid Files

pcDNA™6.2/N-EmGFP-DEST

Vivid Colors™ Gateway® destination vector for expressing N-terminally Emerald GFP-tagged proteins in mammalian cells.

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pcDNA6.2 N-EmGFP-DEST Sequence and MappcDNA6.2 N-EmGFP-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 MfeI (31) SgrDI (7396) AhdI (6475) PciI (5582) BspQI - SapI (5466) BlpI (4910) PpuMI (4674) BclI * (4437) AvrII (4386) StuI (4385) SexAI * (4153) SpeI (119) NdeI (354) SnaBI (460) Eco53kI (686) SacI (688) XbaI (778) EcoRV (800) PspXI (1532) NotI (1671) EcoRI (1991) PflMI * (2223) BssHII (2485) BbvCI (2716) SrfI (2862) BmgBI (2896) BstXI (2979) BfuAI - BspMI (3106) PstI (3117) PmeI (3320) pcDNA™6.2/N-EmGFP-DEST 7528 bp
MfeI  (31)
1 site
C A A T T G G T T A A C
SgrDI  (7396)
1 site
C G T C G A C G G C A G C T G C
AhdI  (6475)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PciI  (5582)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (5466)
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  (5466)
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.
BlpI  (4910)
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.
PpuMI  (4674)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BclI  (4437)
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.
AvrII  (4386)
1 site
C C T A G G G G A T C C
StuI  (4385)
1 site
A G G C C T T C C G G A
SexAI  (4153)
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.
SpeI  (119)
1 site
A C T A G T T G A T C A
NdeI  (354)
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  (460)
1 site
T A C G T A A T G C A T
Eco53kI  (686)
1 site
G A G C T C C T C G A G
SacI  (688)
1 site
G A G C T C C T C G A G
XbaI  (778)
1 site
T C T A G A A G A T C T
EcoRV  (800)
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.
PspXI  (1532)
1 site
V C T C G A G B B G A G C T C V
NotI  (1671)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (1991)
1 site
G A A T T C C T T A A G
PflMI  (2223)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BssHII  (2485)
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.
BbvCI  (2716)
1 site
C C T C A G C G G A G T C G
SrfI  (2862)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (2896)
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.
BstXI  (2979)
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.
BfuAI  (3106)
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  (3106)
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.
PstI  (3117)
1 site
C T G C A G G A C G T C
PmeI  (3320)
1 site
G T T T A A A C C A A A T T T G
AmpR
6402 .. 7262  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6402 .. 7193  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6402 .. 7262  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7194 .. 7262  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6402 .. 7262  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
EmGFP
813 .. 1529  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 1:  
   813 .. 815  =  3 bp
   1 amino acid  =  149.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
813 .. 1529  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 2:  1a  
   816 .. 818  =  3 bp
   1 amino acid  =  117.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
813 .. 1529  =  717 bp
239 amino acids  =  26.9 kDa
   Segment 3:  
   819 .. 1529  =  711 bp
   237 amino acids  =  26.7 kDa
Product: Emerald GFP
mammalian codon-optimized
EmGFP
813 .. 1529  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: Emerald GFP
mammalian codon-optimized
CmR
1778 .. 2437  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1778 .. 2437  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5643 .. 6231  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5643 .. 6231  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3632 .. 4058  =  427 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3632 .. 4058  =  427 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
4515 .. 4913  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4515 .. 4913  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
105 .. 484  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
105 .. 484  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
4072 .. 4401  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4072 .. 4401  =  330 bp
SV40 enhancer and early promoter
ccdB
2779 .. 3084  =  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
2779 .. 3084  =  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
485 .. 688  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
485 .. 688  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
attR1
1545 .. 1669  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1545 .. 1669  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3125 .. 3249  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3125 .. 3249  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5071 .. 5192  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5071 .. 5192  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7263 .. 7367  =  105 bp
AmpR promoter
7263 .. 7367  =  105 bp
HSV TK poly(A) signal
3382 .. 3430  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3382 .. 3430  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
4449 .. 4496  =  48 bp
synthetic bacterial promoter
EM7 promoter
4449 .. 4496  =  48 bp
synthetic bacterial promoter
V5 tag
3257 .. 3298  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3257 .. 3298  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
1694 .. 1724  =  31 bp
   Segment 1:  -35  
   1694 .. 1699  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1694 .. 1724  =  31 bp
   Segment 2:  
   1700 .. 1717  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1694 .. 1724  =  31 bp
   Segment 3:  -10  
   1718 .. 1724  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1694 .. 1724  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
SV40 ori
4252 .. 4387  =  136 bp
SV40 origin of replication
SV40 ori
4252 .. 4387  =  136 bp
SV40 origin of replication
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