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pcDNA™6.2/C-EmGFP-GW TOPO® (linearized)

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

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pcDNA6.2 C-EmGFP-GW TOPO (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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SgrDI (4859) SspI (4742) ScaI (4418) AhdI (3938) PciI (3045) BspQI - SapI (2929) BstZ17I (2666) BsmI (2614) BsaBI * (2276) PpuMI (2137) EagI (2094) BbsI (2063) MscI (1981) BclI * (1900) SmaI (1872) BmeT110I (1871) AvaI - BsoBI - TspMI - XmaI (1870) MfeI (5022) MluI (5089) SpeI (5110) NdeI (5345) SnaBI (5451) Eco53kI (5677) SacI (5679) End (5814) Start (1) HpaI (48) PmeI (781) SexAI * (1616) StuI (1848) AvrII (1849) pcDNA™6.2/C-EmGFP-GW TOPO® 5813 bp
SgrDI  (4859)
1 site
C G T C G A C G G C A G C T G C
SspI  (4742)
1 site
A A T A T T T T A T A A
ScaI  (4418)
1 site
A G T A C T T C A T G A
AhdI  (3938)
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  (3045)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2929)
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  (2929)
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  (2666)
1 site
G T A T A C C A T A T G
BsmI  (2614)
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.
BsaBI  (2276)
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  (2137)
1 site
R G G W C C Y Y C C