pBiEx-3

Vector for high-level expression in both bacterial and insect cells of proteins with a C-terminal S-Tag-8xHis cassette.

Sequence Author: MilliporeSigma (Novagen)

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TspMI - XmaI (5368) BspQI - SapI (5124) PciI (5007) DrdI (4905) AlwNI (4598) AhdI (4119) BsaI (4053) ScaI (3638) XmnI (3519) EcoNI (3130) PfoI (3094) BstAPI (2986) SmaI (5370) BspDI * - ClaI * (207) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BseRI (781) BsiWI (784) SacII (1085) T7 promoter NcoI (1177) ATG BmgBI (1194) Eco53kI (1202) SacI (1204) BamHI (1208) MfeI (1214) BglII (1222) BfuAI - BspMI (1229) AscI (1230) PstI - SbfI (1240) SalI (1242) AccI (1243) Acc65I (1248) AgeI (1251) KpnI (1252) BstBI (1257) HindIII (1260) NotI (1267) PaeR7I - PspXI - XhoI (1344) DraIII (1376) BbvCI - Bpu10I (1386) BlpI (1423) EcoO109I (1450) Bsu36I (1500) PacI (1529) PflFI - Tth111I (1753) KasI (2023) NarI * (2024) SfoI (2025) PluTI (2027) BsmBI - Esp3I (2047) PspOMI (2455) ApaI (2459) BstEII (2480) pBiEx™-3 5373 bp
TspMI  (5368)
1 site
C C C G G G G G G C C C
XmaI  (5368)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
BspQI  (5124)
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  (5124)
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  (5007)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (4905)
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.
AlwNI  (4598)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (4119)
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  (4053)
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  (3638)
1 site
A G T A C T T C A T G A
XmnI  (3519)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoNI  (3130)
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  (3094)
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  (2986)
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.
SmaI  (5370)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
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
BmgBI  (1194)
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.
Eco53kI  (1202)
1 site
G A G C T C C T C G A G
SacI  (1204)
1 site
G A G C T C C T C G A G
BamHI  (1208)
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  (1214)
1 site
C A A T T G G T T A A C
BglII  (1222)
1 site
A G A T C T T C T A G A
BfuAI  (1229)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1229)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
AscI  (1230)
1 site
G G C G C G C C C C G C G C G G
PstI  (1240)
1 site
C T G C A G G A C G T C
SbfI  (1240)
1 site
C C T G C A G G G G A C G T C C
SalI  (1242)
1 site
G T C G A C C A G C T G
AccI  (1243)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
Acc65I  (1248)
1 site
G G T A C C C C A T G G
AgeI  (1251)
1 site
A C C G G T T G G C C A
KpnI  (1252)
1 site
G G T A C C C C A T G G
BstBI  (1257)
1 site
T T C G A A A A G C T T
HindIII  (1260)
1 site
A A G C T T T T C G A A
NotI  (1267)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1344)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.