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pcDNA™6.2/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.

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pcDNA6.2 N-EmGFP-GW TOPO (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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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) MfeI (4350) SpeI (4438) NdeI (4673) SnaBI (4779) Eco53kI (5005) SacI (5007) BsmBI (5028) BspEI (5090) XbaI (5097) BamHI (5103) EcoRV (5119) PspXI (5851) End (5902) Start (1) PmeI (109) SexAI * (944) StuI (1176) AvrII (1177) TspMI - XmaI (1198) 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.
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