pBiEx-1

Vector for high-level expression in both bacterial and insect cells of proteins with a cleavable N-terminal 6xHis-S-Tag cassette plus a C-terminal HSV tag.

Sequence Author: MilliporeSigma (Novagen)

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BspQI - SapI (5226) PciI (5109) DrdI (5007) AhdI (4221) BsaI (4155) ScaI (3740) XmnI (3621) EcoNI (3232) PfoI (3196) BstAPI (3088) BstEII (2582) BspDI * - ClaI * (207) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BseRI (781) BsiWI (784) SacII (1085) lac operator RBS NcoI (1177) ATG 6xHis enterokinase site BglII (1321) SrfI (1333) BamHI (1340) MfeI (1346) Eco53kI (1355) SacI (1357) BsrGI (1360) AscI (1368) PstI - SbfI (1378) SalI (1380) Acc65I (1386) AgeI (1389) KpnI (1390) BstBI (1395) HindIII (1398) NotI (1405) BstZ17I (1420) PmlI (1425) PaeR7I - XhoI (1467) DraIII (1478) BbvCI - Bpu10I (1488) BlpI (1525) EcoO109I (1552) Bsu36I (1602) PacI (1631) PflFI - Tth111I (1855) KasI (2125) NarI * (2126) SfoI (2127) PluTI (2129) BsmBI - Esp3I (2149) HpaI (2262) PspOMI (2557) ApaI (2561) pBiEx™-1 5475 bp
BspQI  (5226)
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  (5226)
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.
PciI  (5109)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (5007)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AhdI  (4221)
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.
BsaI  (4155)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
ScaI  (3740)
1 site
A G T A C T T C A T G A
XmnI  (3621)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoNI  (3232)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PfoI  (3196)
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.
BstAPI  (3088)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BstEII  (2582)
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 typically used at 60°C, but is 50% active at 37°C.
BspDI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PsiI  (409)
1 site
T T A T A A A A T A T T
NheI  (606)
1 site
G C T A G C C G A T C G
BmtI  (610)
1 site
G C T A G C C G A T C G
SphI  (614)
1 site
G C A T G C C G T A C G
BsmI  (775)
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.
BseRI  (781)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BsiWI  (784)
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.
SacII  (1085)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
NcoI  (1177)
1 site
C C A T G G G G T A C C
BglII  (1321)
1 site
A G A T C T T C T A G A
SrfI  (1333)
1 site
G C C C G G G C C G G G C C C G
BamHI  (1340)
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.
MfeI  (1346)
1 site
C A A T T G G T T A A C