pIB V5-His-DEST

Gateway® destination vector for stable constitutive expression of C-terminally V5-6xHis-tagged proteins in insect cells.

Sequence Author: Thermo Fisher (Invitrogen)

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AhdI (4870) BglI (4752) XmnI (4270) EarI (4071) BglII (4003) DraIII (3970) BlpI (3960) NruI * (3796) PpuMI (3724) MreI - SgrAI (3658) BbsI (3650) SpeI (3415) BciVI (2826) PciI (2623) BspDI * - ClaI * (2560) BspHI (0) FseI (293) BsiWI (317) RsrII (335) KasI (474) NarI (475) SfoI (476) PluTI (478) HindIII (557) EcoRV (565) AfeI (586) NotI (728) BspEI (1044) BsmBI - Esp3I (1272) PasI - PflMI * (1280) SspI (1360) BssHII (1542) BbvCI (1773) SrfI (1919) BstXI (2036) BfuAI - BspMI (2163) SalI (2176) PshAI (2315) XbaI (2319) SacII (2332) AgeI (2386) 6xHis PsiI (2493) pIB/V5-His-DEST 5038 bp
AhdI  (4870)
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.
BglI  (4752)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
XmnI  (4270)
1 site
G A A N N N N T T C C T T N N N N A A G
EarI  (4071)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
BglII  (4003)
1 site
A G A T C T T C T A G A
DraIII  (3970)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BlpI  (3960)
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.
NruI  (3796)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
PpuMI  (3724)
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.
MreI  (3658)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (3658)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BbsI  (3650)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
SpeI  (3415)
1 site
A C T A G T T G A T C A
BciVI  (2826)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
PciI  (2623)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspDI  (2560)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2560)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BspHI  (0)
1 site
T C A T G A A G T A C T
FseI  (293)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BsiWI  (317)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
RsrII  (335)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
KasI  (474)
1 site
G G C G C C C C G C G G
NarI  (475)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (476)
1 site
G G C G C C C C G C G G
PluTI  (478)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
HindIII  (557)
1 site
A A G C T T T T C G A A
EcoRV  (565)
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.
AfeI  (586)
1 site
A G C G C T T C G C G A
NotI  (728)
1 site
G C G G C C G C C G C C G G C G
BspEI  (1044)
1 site
T C C G G A A G G C C T
BsmBI  (1272)
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  (1272)
1 site
C G T C T C N G