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Plasmid Files

pET-40b(+)

Bacterial vector for expressing periplasmic DsbC fusion proteins.

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pET-40b(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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AvrII (131) DraIII (5948) AsiSI - PvuI (5247) NruI (4904) AcuI (4593) BssS α I (4218) PciI (4045) BspQI - SapI (3929) BstZ17I (3816) PflFI - Tth111I (3790) PaeR7I - PspXI - XhoI (174) EagI - NotI (182) HindIII (189) SalI (195) Eco53kI (204) SacI (206) EcoRI (208) BamHI (214) EcoRV (222) NcoI (227) SrfI (246) BseRI (250) Acc65I (267) KpnI (271) BglII (274) BstBI (301) MfeI (332) ScaI (341) SacII (365) SpeI (396) BfuAI - BspMI (600) BsrGI (941) NdeI (1114) XbaI (1152) SgrAI (1263) SphI (1419) BstAPI (1627) MluI (1944) BclI * (1958) NmeAIII (2150) PspOMI (2151) ApaI (2155) BssHII (2355) HpaI (2450) PshAI (2789) BglI (3008) FspI - FspAI (3026) PpuMI (3051) pET-40b(+) 6190 bp
AvrII  (131)
1 site
C C T A G G G G A T C C
DraIII  (5948)
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.
AsiSI  (5247)
1 site
G C G A T C G C C G C T A G C G
PvuI  (5247)
1 site
C G A T C G G C T A G C
NruI  (4904)
1 site
T C G C G A A G C G C T
AcuI  (4593)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
BssSαI  (4218)
1 site
C A C G A G G T G C T C
PciI  (4045)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3929)
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  (3929)
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  (3816)
1 site
G T A T A C C A T A T G
PflFI  (3790)
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  (3790)
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.
PaeR7I  (174)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (174)
1 site
V C T C G A G B B G A G C T C V
XhoI  (174)
1 site
C T C G A G G A G C T C
EagI  (182)
1 site
C G G C C G G C C G G C
NotI  (182)
1 site
G C G G C C G C C G C C G G C G
HindIII  (189)
1 site
A A G C T T T T C G A A
SalI  (195)
1 site
G T C G A C C A G C T G
Eco53kI  (204)
1 site
G A G C T C C T C G A G
SacI  (206)
1 site
G A G C T C C T C G A G
EcoRI  (208)
1 site
G A A T T C C T T A A G
BamHI  (214)
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.
EcoRV  (222)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
NcoI  (227)
1 site
C C A T G G G G T A C C
SrfI  (246)
1 site
G C C C G G G C C G G G C C C G
BseRI  (250)
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.
Acc65I  (267)
1 site
G G T A C C C C A T G G
KpnI  (271)
1 site
G G T A C C C C A T G G
BglII  (274)
1 site
A G A T C T T C T A G A
BstBI  (301)
1 site
T T C G A A A A G C T T
MfeI  (332)
1 site
C A A T T G G T T A A C
ScaI  (341)
1 site
A G T A C T T C A T G A
SacII  (365)
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.
SpeI  (396)
1 site
A C T A G T T G A T C A
BfuAI  (600)
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  (600)
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  (941)
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.
NdeI  (1114)
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  (1152)
1 site
T C T A G A A G A T C T
SgrAI  (1263)
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  (1419)
1 site
G C A T G C C G T A C G
BstAPI  (1627)
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  (1944)
1 site
A C G C G T T G C G C A
BclI  (1958)
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.
NmeAIII  (2150)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PspOMI  (2151)
1 site
G G G C C C C C C G G G
ApaI  (2155)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BssHII  (2355)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
HpaI  (2450)
1 site
G T T A A C C A A T T G
PshAI  (2789)
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.
BglI  (3008)
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  (3026)
1 site
T G C G C A A C G C G T
FspAI  (3026)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (3051)
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.
lacI
1594 .. 2676  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
1594 .. 2676  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
KanR
4816 .. 5631  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
4816 .. 5631  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
DsbC
408 .. 1115  =  708 bp
236 amino acids  =  25.6 kDa
Product: bacterial periplasmic protein disulfide isomerase
DsbC
408 .. 1115  =  708 bp
236 amino acids  =  25.6 kDa
Product: bacterial periplasmic protein disulfide isomerase
ori
4106 .. 4694  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4106 .. 4694  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5724 .. 6179  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis