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pET-45b(+)

Bacterial vector for expressing N-terminally 6xHis-tagged and C-terminally S-tagged proteins.

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pET-45b(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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DraIII (5018) PsiI (4890) SspI (4810) AhdI (4606) BsaI (4540) BglI (4488) FspI (4383) PvuI (4237) ScaI (4125) AlwNI (3412) PciI (2996) BspQI - SapI (2880) BstZ17I (2767) PflFI - Tth111I (2741) BlpI (80) AvrII (97) PacI (107) AvaI - BsoBI - PaeR7I - PspXI - XhoI (176) BmeT110I (177) EagI - NotI (184) HindIII (191) SalI (197) PstI - SbfI (207) AscI (209) BfuAI - BspMI (210) BsrGI (217) Eco53kI (226) SacI (228) MfeI (231) BamHI (237) PshAI (248) BstBI (265) AgeI (269) Acc65I (272) KpnI (276) PmlI (280) NcoI (301) XbaI (340) BspDI * - ClaI * (409) SgrAI (451) SphI (607) EcoNI (667) BstAPI (815) MluI (1132) BclI * (1146) BstEII (1313) PspOMI (1339) ApaI (1343) HpaI (1638) PpuMI (2002) Bpu10I (2102) pET-45b(+) 5260 bp
DraIII  (5018)
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.
PsiI  (4890)
1 site
T T A T A A A A T A T T
SspI  (4810)
1 site
A A T A T T T T A T A A
AhdI  (4606)
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  (4540)
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.
BglI  (4488)
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.
FspI  (4383)
1 site
T G C G C A A C G C G T
PvuI  (4237)
1 site
C G A T C G G C T A G C
ScaI  (4125)
1 site
A G T A C T T C A T G A
AlwNI  (3412)
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.
PciI  (2996)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2880)
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  (2880)
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  (2767)
1 site
G T A T A C C A T A T G
PflFI  (2741)
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  (2741)
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.
BlpI  (80)
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.
AvrII  (97)
1 site
C C T A G G G G A T C C
PacI  (107)
1 site
T T A A T T A A A A T T A A T T
AvaI  (176)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (176)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (176)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (176)
1 site
V C T C G A G B B G A G C T C V
XhoI  (176)
1 site
C T C G A G G A G C T C
BmeT110I  (177)
1 site
C Y C G R G G R G C Y C
EagI  (184)
1 site
C G G C C G G C C G G C
NotI  (184)
1 site
G C G G C C G C C G C C G G C G
HindIII  (191)
1 site
A A G C T T T T C G A A
SalI  (197)
1 site
G T C G A C C A G C T G
PstI  (207)
1 site
C T G C A G G A C G T C
SbfI  (207)
1 site
C C T G C A G G G G A C G T C C
AscI  (209)
1 site
G G C G C G C C C C G C G C G G
BfuAI  (210)
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  (210)
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.
BsrGI  (217)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
Eco53kI  (226)
1 site
G A G C T C C T C G A G
SacI  (228)
1 site
G A G C T C C T C G A G
MfeI  (231)
1 site
C A A T T G G T T A A C
BamHI  (237)
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.
PshAI  (248)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BstBI  (265)
1 site
T T C G A A A A G C T T
AgeI  (269)
1 site
A C C G G T T G G C C A
Acc65I  (272)
1 site
G G T A C C C C A T G G
KpnI  (276)
1 site