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pET-25b(+)

Bacterial vector with a signal sequence for expression of C-terminally HSV-tagged proteins in the periplasm.

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pET-25b(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  MilliporeSigma (Novagen)
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DraIII (5305) PsiI (5177) ScaI (4642) PvuI (4532) PstI (4407) BsaI (4223) AhdI (4162) AlwNI (3685) PciI (3269) BspQI - SapI (3153) BstZ17I (3040) PflFI - Tth111I (3014) BlpI (80) NheI (197) BmtI (201) PaeR7I - XhoI (212) EagI - NotI (220) HindIII (227) SalI (233) Eco53kI (242) SacI (244) EcoRI (246) BamHI (252) NcoI (274) MscI (279) BseRI (314) BfuAI - BspMI (322) NdeI (342) XbaI (380) BglII (446) SgrAI (487) SphI (643) EcoNI (703) PflMI (750) BstAPI (851) MluI (1168) BclI * (1182) BstEII (1349) PspOMI (1375) ApaI (1379) BssHII (1579) HpaI (1674) PshAI (2013) FspAI (2250) PpuMI (2275) Bpu10I (2375) pET-25b(+) 5547 bp
DraIII  (5305)
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  (5177)
1 site
T T A T A A A A T A T T
ScaI  (4642)
1 site
A G T A C T T C A T G A
PvuI  (4532)
1 site
C G A T C G G C T A G C
PstI  (4407)
1 site
C T G C A G G A C G T C
BsaI  (4223)
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.
AhdI  (4162)
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.
AlwNI  (3685)
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  (3269)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3153)
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  (3153)
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  (3040)
1 site
G T A T A C C A T A T G
PflFI  (3014)
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  (3014)
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.
NheI  (197)
1 site
G C T A G C C G A T C G
BmtI  (201)
1 site
G C T A G C C G A T C G
PaeR7I  (212)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (212)
1 site
C T C G A G G A G C T C
EagI  (220)
1 site
C G G C C G G C C G G C
NotI  (220)
1 site
G C G G C C G C C G C C G G C G
HindIII  (227)
1 site
A A G C T T T T C G A A
SalI  (233)
1 site
G T C G A C C A G C T G
Eco53kI  (242)
1 site
G A G C T C C T C G A G
SacI  (244)
1 site
G A G C T C C T C G A G
EcoRI  (246)
1 site
G A A T T C C T T A A G
BamHI  (252)
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.
NcoI  (274)
1 site
C C A T G G G G T A C C
MscI  (279)
1 site
T G G C C A A C C G G T
BseRI  (314)
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.
BfuAI  (322)
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  (322)
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.
NdeI  (342)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
XbaI  (380)
1 site
T C T A G A A G A T C T
BglII  (446)
1 site
A G A T C T T C T A G A
SgrAI  (487)
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.
SphI  (643)
1 site
G C A T G C C G T A C G
EcoNI  (703)
1 site
C C T N N N N N A G G G G A N N N N N T C C