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pETDuet™-1

Bacterial vector for the co-expression of two genes.

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pETDuet-1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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EcoRI (112) BamHI (106) NcoI (69) XbaI (30) BspDI * - ClaI * (5383) pET Upstream Primer (5358 .. 5373) SgrAI (5339) SphI (5191) EcoNI (5126) BstAPI (4982) MluI (4658) BclI * (4644) BstEII (4476) ApaI (4455) PspOMI (4451) HpaI (4156) PpuMI (3789) Bpu10I (3689) PflFI - Tth111I (3052) BstZ17I (3027) BspQI - SapI (2911) PciI (2794) Eco53kI (120) SacI (122) BfuAI - BspMI (124) AscI (125) PstI - SbfI (135) SalI (137) HindIII (143) NotI (150) AflII (163) DuetUP2 Primer (189 .. 208) BsrGI (190) DuetDOWN1 Primer (189 .. 208) NdeI (298) BglII (305) MfeI (311) EcoRV (319) FseI (328) AsiSI (337) ZraI (344) AatII (346) Acc65I (348) KpnI (352) AvaI - BsoBI - PaeR7I - PspXI - XhoI (354) BmeT110I (355) PacI (429) AvrII (433) BlpI (451) T7 Terminator Primer (448 .. 466) DraIII (779) PsiI (904) SspI (984) AhdI (1189) BsaI (1250) BglI (1309) FspI (1411) ScaI (1669) AlwNI (2385) pETDuet™-1 5420 bp
EcoRI  (112)
1 site
G A A T T C C T T A A G
BamHI  (106)
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  (69)
1 site
C C A T G G G G T A C C
XbaI  (30)
1 site
T C T A G A A G A T C T
BspDI  (5383)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (5383)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (5339)
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  (5191)
1 site
G C A T G C C G T A C G
EcoNI  (5126)
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.
BstAPI  (4982)
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.
MluI  (4658)
1 site
A C G C G T T G C G C A
BclI  (4644)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (4476)
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.
ApaI  (4455)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4451)
1 site
G G G C C C C C C G G G
HpaI  (4156)
1 site
G T T A A C C A A T T G
PpuMI  (3789)
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.
Bpu10I  (3689)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
PflFI  (3052)
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  (3052)
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.
BstZ17I  (3027)
1 site
G T A T A C C A T A T G
BspQI  (2911)
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  (2911)
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  (2794)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
Eco53kI  (120)
1 site
G A G C T C C T C G A G
SacI  (122)
1 site
G A G C T C C T C G A G
BfuAI  (124)
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  (124)
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  (125)
1 site
G G C G C G C C C C G C G C G G
PstI  (135)
1 site
C T G C A G G