pcDNA6.2 N-EmGFP-DEST

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspcDNA6.2 N-EmGFP-DEST
MfeI (31) SgrDI (7396) AhdI (6475) PciI (5582) BspQI - SapI (5466) BlpI (4910) PpuMI (4674) BclI * (4437) AvrII (4386) StuI (4385) CsiI - 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
CsiI  (4153)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
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
2 segments
   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
2 segments
   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
3 segments
   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
3 segments
   Segment 2:  1a  
   816 .. 818  =  3 bp
   1 amino acid  =  117.1 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
813 .. 1529  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   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
3 segments
   Segment 1:  -35  
   1694 .. 1699  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1694 .. 1724  =  31 bp
3 segments
   Segment 2:  
   1700 .. 1717  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1694 .. 1724  =  31 bp
3 segments
   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
ORF:  2779 .. 3084  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  6532 .. 6798  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1778 .. 2437  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  813 .. 1580  =  768 bp
ORF:  255 amino acids  =  28.7 kDa
ORF:  4515 .. 4913  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  4461 .. 4817  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  6402 .. 7262  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pcDNA6.2 N-EmGFP-DEST.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

Plasmid Sets

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