pcDNA6.2 N-EmGFP-GW TOPO (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning to add an N-terminal GFP tag and create a Gateway® expression clone.

Sequence Author: Thermo Fisher (Invitrogen)

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: TA and GC Cloning Vectors | More Plasmid Sets
No matches
SgrDI (4187) SspI (4070) ScaI (3746) AhdI (3266) PciI (2373) BspQI - SapI (2257) BstZ17I (1994) BsmI (1942) BlpI (1701) BsaBI * (1604) PpuMI (1465) EagI (1422) MscI (1309) BclI * (1228) SmaI (1200) TspMI - XmaI (1198) MfeI (4350) SpeI (4438) NdeI (4673) SnaBI (4779) Eco53kI (5005) SacI (5007) BsmBI - Esp3I (5028) BspEI (5090) XbaI (5097) BamHI (5103) EcoRV (5119) PspXI (5851) End (5902) Start (1) attB2 PmeI (109) CsiI - SexAI * (944) StuI (1176) AvrII (1177) pcDNA™6.2/N-EmGFP-GW TOPO® 5901 bp
SgrDI  (4187)
1 site
C G T C G A C G G C A G C T G C
SspI  (4070)
1 site
A A T A T T T T A T A A
ScaI  (3746)
1 site
A G T A C T T C A T G A
AhdI  (3266)
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  (2373)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2257)
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  (2257)
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.
BstZ17I  (1994)
1 site
G T A T A C C A T A T G
BsmI  (1942)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BlpI  (1701)
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.
BsaBI  (1604)
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.
PpuMI  (1465)
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.
EagI  (1422)
1 site
C G G C C G G C C G G C
MscI  (1309)
1 site
T G G C C A A C C G G T
BclI  (1228)
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.
SmaI  (1200)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (1198)
1 site
C C C G G G G G G C C C
XmaI  (1198)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
MfeI  (4350)
1 site
C A A T T G G T T A A C
SpeI  (4438)
1 site
A C T A G T T G A T C A
NdeI  (4673)
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  (4779)
1 site
T A C G T A A T G C A T
Eco53kI  (5005)
1 site
G A G C T C C T C G A G
SacI  (5007)
1 site
G A G C T C C T C G A G
BsmBI  (5028)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (5028)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BspEI  (5090)
1 site
T C C G G A A G G C C T
XbaI  (5097)
1 site
T C T A G A A G A T C T
BamHI  (5103)
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.
EcoRV  (5119)
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  (5851)
1 site
V C T C G A G B B G A G C T C V
End  (5902)
0 sites
Start  (1)
0 sites
PmeI  (109)
1 site
G T T T A A A C C A A A T T T G
CsiI  (944)
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  (944)
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.
StuI  (1176)
1 site
A G G C C T T C C G G A
AvrII  (1177)
1 site
C C T A G G G G A T C C
AmpR
3193 .. 4053  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3193 .. 3984  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3193 .. 4053  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3985 .. 4053  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3193 .. 4053  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
EmGFP
5132 .. 5848  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   5132 .. 5134  =  3 bp
   1 amino acid  =  149.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
5132 .. 5848  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   5135 .. 5137  =  3 bp
   1 amino acid  =  117.1 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
5132 .. 5848  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   5138 .. 5848  =  711 bp
   237 amino acids  =  26.7 kDa
Product: Emerald GFP
mammalian codon-optimized
EmGFP
5132 .. 5848  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: Emerald GFP
mammalian codon-optimized
ori
2434 .. 3022  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
2434 .. 3022  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
f1 ori
421 .. 849  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
421 .. 849  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
1306 .. 1704  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
1306 .. 1704  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
4424 .. 4803  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4424 .. 4803  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
863 .. 1192  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
863 .. 1192  =  330 bp
SV40 enhancer and early promoter
CMV promoter
4804 .. 5007  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
4804 .. 5007  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
1862 .. 1983  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1862 .. 1983  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4054 .. 4158  =  105 bp
AmpR promoter
4054 .. 4158  =  105 bp
HSV TK poly(A) signal
171 .. 219  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
171 .. 219  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
1240 .. 1287  =  48 bp
synthetic bacterial promoter
EM7 promoter
1240 .. 1287  =  48 bp
synthetic bacterial promoter
V5 tag
46 .. 87  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
46 .. 87  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
attB2
14 .. 38  =  25 bp
recombination site for the Gateway® BP reaction
attB2
14 .. 38  =  25 bp
recombination site for the Gateway® BP reaction
attB1
5864 .. 5888  =  25 bp
recombination site for the Gateway® BP reaction
attB1
5864 .. 5888  =  25 bp
recombination site for the Gateway® BP reaction
SV40 ori
1043 .. 1178  =  136 bp
SV40 origin of replication
SV40 ori
1043 .. 1178  =  136 bp
SV40 origin of replication
ORF:  1306 .. 1704  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  3323 .. 3589  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5132 .. 5902  =  771 bp
ORF:  257 amino acids  =  28.7 kDa
ORF:  1252 .. 1608  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  3193 .. 4053  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
Click here to try SnapGene

Download pcDNA6.2 N-EmGFP-GW TOPO (linearized).dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps

The maps, notes, and annotations in the zip file on this page are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be