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pMIB/V5-His C

Insect cell vector for expression of secreted, C-terminally V5-6xHis-tagged proteins. For other reading frames, use pMIB/V5‑His A or pMIB/V5‑His B.

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pMIB V5-His C.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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AhdI (3424) BpmI (3355) BglI (3306) ScaI (2943) XmnI (2824) EarI (2625) BglII (2557) SmaI (2553) TspMI - XmaI (2551) BmgBI (2548) DraIII (2524) BsaBI * (2417) NruI * (2350) PvuII (2326) EcoO109I - PpuMI (2278) BbsI (2204) MscI (2122) BtgI - NcoI - StyI (2117) PmlI (2050) BspHI (0) PspOMI (150) ApaI (154) BsiWI (317) RsrII (335) KasI (474) NarI (475) SfoI (476) PluTI (478) SphI (633) HindIII (636) Acc65I (642) KpnI (646) Eco53kI (650) SacI (652) BamHI (654) SpeI (660) EcoRI (677) EcoRV (689) NotI (704) PaeR7I - PspXI - XhoI (710) BstEII (715) AgeI (775) PsiI (882) BspDI * - ClaI * (949) PciI (1012) BciVI (1215) pMIB/V5-His C 3592 bp
AhdI  (3424)
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.
BpmI  (3355)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BglI  (3306)
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.
ScaI  (2943)
1 site
A G T A C T T C A T G A
XmnI  (2824)
1 site
G A A N N N N T T C C T T N N N N A A G
EarI  (2625)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI recognition sequence.
Sticky ends from different EarI sites may not be compatible.
BglII  (2557)
1 site
A G A T C T T C T A G A
SmaI  (2553)
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  (2551)
1 site
C C C G G G G G G C C C
XmaI  (2551)
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.
BmgBI  (2548)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
DraIII  (2524)
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.
BsaBI  (2417)
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.
NruI  (2350)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
PvuII  (2326)
1 site
C A G C T G G T C G A C
EcoO109I  (2278)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PpuMI  (2278)
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.
BbsI  (2204)
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.
MscI  (2122)
1 site
T G G C C A A C C G G T
BtgI  (2117)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (2117)
1 site
C C A T G G G G T A C C
StyI  (2117)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PmlI  (2050)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BspHI  (0)
1 site
T C A T G A A G T A C T
PspOMI  (150)
1 site
G G G C C C C C C G G G
ApaI  (154)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°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.
SphI  (633)
1 site
G C A T G C C G T A C G
HindIII  (636)
1 site
A A G C T T T T C G A A
Acc65I  (642)
1 site
G G T A C C C C A T G G
KpnI  (646)
1 site
G G T A C C C C A T G G
Eco53kI  (650)
1 site
G A G C T C C T C G A G
SacI  (652)
1 site
G A G C T C C T C G A G
BamHI  (654)
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.
SpeI  (660)
1 site
A C T A G T T G A T C A
EcoRI  (677)
1 site
G A A T T C C T T A A G
EcoRV  (689)
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.
NotI  (704)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (710)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (710)
1 site
V C T C G A G B B G A G C T C V
XhoI  (710)
1 site
C T C G A G G A G C T C
BstEII  (715)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typ