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Plasmid Files

pcDNA™6.2/C-EmGFP-DEST

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

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pcDNA6.2 C-EmGFP-DEST Sequence and MappcDNA6.2 C-EmGFP-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 MfeI (31) SgrDI (7310) AhdI (6389) PciI (5496) BspQI - SapI (5380) PpuMI (4588) BbsI (4514) BclI * (4351) AvrII (4300) StuI (4299) SexAI * (4067) MluI (98) SpeI (119) CMV enhancer NdeI (354) SnaBI (460) Eco53kI (686) SacI (688) NotI (911) lac UV5 promoter BspEI (1227) EcoRI (1231) BsmBI (1455) PflMI * (1463) BssHII (1725) BbvCI (1956) SrfI (2102) BmgBI (2136) BstXI (2219) BfuAI - BspMI (2346) PstI (2357) HpaI (2499) PmeI (3232) pcDNA™6.2/C-EmGFP-DEST 7442 bp
MfeI  (31)
1 site
C A A T T G G T T A A C
SgrDI  (7310)
1 site
C G T C G A C G G C A G C T G C
AhdI  (6389)
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  (5496)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (5380)
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  (5380)
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.
PpuMI  (4588)
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.
BbsI  (4514)
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.
BclI  (4351)
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  (4300)
1 site
C C T A G G G G A T C C
StuI  (4299)
1 site
A G G C C T T C C G G A
SexAI  (4067)
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.
MluI  (98)
1 site
A C G C G T T G C G C A
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
NotI  (911)
1 site
G C G G C C G C C G C C G G C G
BspEI  (1227)
1 site
T C C G G A A G G C C T
EcoRI  (1231)
1 site
G A A T T C C T T A A G
BsmBI  (1455)
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.
PflMI  (1463)
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  (1725)
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  (1956)
1 site
C C T C A G C G G A G T C G
SrfI  (2102)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (2136)
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  (2219)
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  (2346)
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  (2346)
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  (2357)
1 site
C T G C A G G A C G T C
HpaI  (2499)
1 site
G T T A A C C A A T T G
PmeI  (3232)
1 site
G T T T A A A C C A A A T T T G
AmpR
6316 .. 7176  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6316 .. 7107  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6316 .. 7176  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7108 .. 7176  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6316 .. 7176  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
EmGFP
2503 .. 3222  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 1:  
   2503 .. 2505  =  3 bp
   1 amino acid  =  149.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
2503 .. 3222  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 2:  1a  
   2506 .. 2508  =  3 bp
   1 amino acid  =  117.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
2503 .. 3222  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 3:  
   2509 .. 3222  =  714 bp
   237 amino acids  =  26.7 kDa
Product: Emerald GFP
mammalian codon-optimized
EmGFP
2503 .. 3222  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: Emerald GFP
mammalian codon-optimized
CmR
1018 .. 1677  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1018 .. 1677  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5557 .. 6145  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5557 .. 6145  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3544 .. 3972  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3544 .. 3972  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
4429 .. 4827  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4429 .. 4827  =  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
3986 .. 4315  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3986 .. 4315  =  330 bp
SV40 enhancer and early promoter
ccdB
2019 .. 2324  =  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
2019 .. 2324  =  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
785 .. 909  =  125 bp
recombination site for the Gateway® LR reaction
attR1
785 .. 909  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2365 .. 2489  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2365 .. 2489  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
4985 .. 5106  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4985 .. 5106  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7177 .. 7281  =  105 bp
AmpR promoter
7177 .. 7281  =  105 bp
HSV TK poly(A) signal
3294 .. 3342  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3294 .. 3342  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
4363 .. 4410  =  48 bp
synthetic bacterial promoter
EM7 promoter
4363 .. 4410  =  48 bp
synthetic bacterial promoter
lac UV5 promoter
934 .. 964  =  31 bp
   Segment 1:  -35  
   934 .. 939  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
934 .. 964  =  31 bp
   Segment 2:  
   940 .. 957  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
934 .. 964  =  31 bp
   Segment 3:  -10  
   958 .. 964  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
934 .. 964  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
733 .. 751  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
733 .. 751  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
4166 .. 4301  =  136 bp
SV40 origin of replication
SV40 ori
4166 .. 4301  =  136 bp
SV40 origin of replication
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