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Plasmid Files

pET-39b(+)

Bacterial vector for expressing periplasmic DsbA fusion proteins.

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pET-39b(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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 8xHis AvrII (131) T7 terminator DraIII (5864) PsiI (5736) AsiSI - PvuI (5163) NruI (4820) AcuI (4509) AlwNI (4377) BssS α I (4134) PciI (3961) BspQI - SapI (3845) BstZ17I (3732) PflFI - Tth111I (3706) PpuMI (2967) PaeR7I - PspXI - XhoI (174) EagI - NotI (182) HindIII (189) SalI (195) Eco53kI (204) SacI (206) EcoRI (208) BamHI (214) NcoI (227) SrfI (246) BseRI (250) enterokinase site Acc65I (267) KpnI (271) BstBI (301) ScaI (341) thrombin site SacII (365) 6xHis SpeI (396) AflII (418) PstI (552) BfuAI - BspMI (636) AscI (917) AgeI (928) NdeI (1030) RBS XbaI (1068) T7 promoter SgrAI (1179) SphI (1335) BstAPI (1543) MluI (1860) BclI * (1874) NmeAIII (2066) PspOMI (2067) ApaI (2071) PshAI (2705) BglI (2924) FspI - FspAI (2942) pET-39b(+) 6106 bp
AvrII  (131)
1 site
C C T A G G G G A T C C
DraIII  (5864)
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  (5736)
1 site
T T A T A A A A T A T T
AsiSI  (5163)
1 site
G C G A T C G C C G C T A G C G
PvuI  (5163)
1 site
C G A T C G G C T A G C
NruI  (4820)
1 site
T C G C G A A G C G C T
AcuI  (4509)
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).
AlwNI  (4377)
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.
BssSαI  (4134)
1 site
C A C G A G G T G C T C
PciI  (3961)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3845)
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  (3845)
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  (3732)
1 site
G T A T A C C A T A T G
PflFI  (3706)
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  (3706)
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.
PpuMI  (2967)
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.
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.
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
BstBI  (301)
1 site
T T C G A A A A G C T T
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
AflII  (418)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
PstI  (552)
1 site
C T G C A G G A C G T C
BfuAI  (636)
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  (636)
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  (917)
1 site
G G C G C G C C C C G C G C G G
AgeI  (928)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
NdeI  (1030)
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  (1068)
1 site
T C T A G A A G A T C T
SgrAI  (1179)
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  (1335)
1 site
G C A T G C C G T A C G
BstAPI  (1543)
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  (1860)
1 site
A C G C G T T G C G C A
BclI  (1874)
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  (2066)
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  (2067)
1 site
G G G C C C C C C G G G
ApaI  (2071)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PshAI  (2705)
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  (2924)
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  (2942)
1 site
T G C G C A A C G C G T
FspAI  (2942)
1 site
R T G C G C A Y Y A C G C G T R
lacI
1510 .. 2592  =  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
1510 .. 2592  =  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
4732 .. 5547  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
4732 .. 5547  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
DsbA
408 .. 1031  =  624 bp
208 amino acids  =  23.1 kDa
Product: bacterial periplasmic oxidoreductase
DsbA
408 .. 1031  =  624 bp
208 amino acids  =  23.1 kDa
Product: bacterial periplasmic oxidoreductase
ori
4022 .. 4610  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4022 .. 4610  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
5640 .. 6095  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
5640 .. 6095  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rop
3401 .. 3592  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
3401 .. 3592  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
lacI promoter
1432 .. 1509  =  78 bp
lacI promoter
1432 .. 1509  =  78 bp
MCS
174 .. 250  =  77 bp
multiple cloning site
MCS
174 .. 250  =  77 bp
multiple cloning site
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
S-Tag
282 .. 326  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
S-Tag
282 .. 326  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
lac operator
1076 .. 1100  =  25 bp
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).
lac operator
1076 .. 1100  =  25 bp
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).
8xHis
150 .. 173  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
150 .. 173  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
T7 promoter
1101 .. 1119  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1101 .. 1119  =  19 bp
promoter for bacteriophage T7 RNA polymerase
thrombin site
342 .. 359  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
342 .. 359  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
6xHis
369 .. 386  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
369 .. 386  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
enterokinase site
252 .. 266  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
252 .. 266  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
RBS
1040 .. 1045  =  6 bp
ribosome binding site
RBS
1040 .. 1045  =  6 bp
ribosome binding site
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