Resources
Plasmid Files

pEF6/V5-His-TOPO® (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products and expression of tagged proteins in mammalian cells.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pEF6 V5-His-TOPO (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
Download Free Trial Get SnapGene Viewer


SspI (4084) ScaI (3760) FspI (3502) AhdI (3280) BspQI - SapI (2271) BstZ17I (2008) BsmI (1956) PfoI (1811) BlpI (1715) BsaBI * (1618) PflFI - Tth111I (1594) PpuMI (1479) MscI (1323) SmaI (1214) TspMI - XmaI (1212) AvrII (1191) ZraI (4200) SgrDI (4201) AatII (4202) AflII (4935) BfuAI - BspMI (5080) FseI (5260) PasI (5304) BbvCI (5458) KasI (5494) NarI (5495) SfoI (5496) PluTI (5498) Acc65I (5796) KpnI (5800) BamHI (5808) SpeI (5814) End (5841) Start (1) EcoRV (18) NotI (33) PspXI (39) XbaI (45) BstBI (62) PmeI (142) SexAI * (958) pEF6/V5-His-TOPO® 5840 bp
SspI  (4084)
1 site
A A T A T T T T A T A A
ScaI  (3760)
1 site
A G T A C T T C A T G A
FspI  (3502)
1 site
T G C G C A A C G C G T
AhdI  (3280)
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.
BspQI  (2271)
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  (2271)
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  (2008)
1 site
G T A T A C C A T A T G
BsmI  (1956)
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.
PfoI  (1811)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BlpI  (1715)
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  (1618)
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.
PflFI  (1594)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1594)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PpuMI  (1479)
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.
MscI  (1323)
1 site
T G G C C A A C C G G T
SmaI  (1214)
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  (1212)
1 site
C C C G G G G G G C C C
XmaI  (1212)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
AvrII  (1191)
1 site