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pET-26b(+)

Bacterial vector with a signal sequence and a kanamycin resistance marker for expression of proteins in the periplasm.

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pET-26b(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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DraIII (5118) PsiI (4990) AsiSI - PvuI (4417) SmaI (4291) TspMI - XmaI (4289) BspDI - ClaI (4108) NruI (4074) AcuI (3763) AlwNI (3631) BssS α I (3388) PciI (3215) BspQI - SapI (3099) TatI (3019) BstZ17I (2986) PflFI - Tth111I (2960) BlpI (80) PaeR7I - PspXI - XhoI (158) EagI - NotI (166) HindIII (173) SalI (179) Eco53kI (188) SacI (190) EcoRI (192) BamHI (198) NcoI (220) MscI (225) BseRI (260) BfuAI - BspMI (268) NdeI (288) XbaI (326) BglII (392) SgrAI (433) SphI (589) BstAPI (797) MluI (1114) BclI * (1128) BstEII (1295) NmeAIII (1320) PspOMI (1321) ApaI (1325) BssHII (1525) HpaI (1620) PshAI (1959) FspI - FspAI (2196) PpuMI (2221) pET-26b(+) 5360 bp
DraIII  (5118)
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  (4990)
1 site
T T A T A A A A T A T T
AsiSI  (4417)
1 site
G C G A T C G C C G C T A G C G
PvuI  (4417)
1 site
C G A T C G G C T A G C
SmaI  (4291)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (4289)
1 site
C C C G G G G G G C C C
XmaI  (4289)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
BspDI  (4108)
1 site
A T C G A T T A G C T A
ClaI  (4108)
1 site
A T C G A T T A G C T A
NruI  (4074)
1 site
T C G C G A A G C G C T
AcuI  (3763)
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  (3631)
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  (3388)
1 site
C A C G A G G T G C T C
PciI  (3215)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3099)
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  (3099)
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.
TatI  (3019)
1 site
W G T A C W W C A T G W
BstZ17I  (2986)
1 site
G T A T A C C A T A T G
PflFI  (2960)
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  (2960)
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.
PaeR7I  (158)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site
V C T C G A G B B G A G C T C V
XhoI  (158)
1 site
C T C G A G G A G C T C
EagI  (166)
1 site
C G G C C G G C C G G C
NotI  (166)
1 site
G C G G C C G C C G C C G G C G
HindIII  (173)
1 site
A A G C T T T T C G A A
SalI  (179)
1 site
G T C G A C C A G C T G
Eco53kI  (188)
1 site
G A G C T C C T C G A G
SacI  (190)
1 site
G A G C T C C T C G A G
EcoRI  (192)
1 site
G A A T T C C T T A A G
BamHI  (198)
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  (220)
1 site
C C A T G G G G T A C C
MscI  (225)
1 site
T G G C C A A C C G G T
BseRI  (260)
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  (268)
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  (268)
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  (288)
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  (326)
1 site
T C T A G A A G A T C T
BglII  (392)
1 site
A G A T C T T C T A G A
SgrAI  (433)
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  (589)
1 site
G C A T G C C G T A C G
BstAPI  (797)
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  (1114)
1 site
A C G C G T T G C G C A
BclI  (1128)
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  (1295)
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.
NmeAIII  (1320)
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  (1321)
1 site
G G G C C C C C C G G G
ApaI  (1325)
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  (1525)
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  (1620)
1 site
G T T A A C C A A T T G
PshAI  (1959)
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.
FspI  (2196)
1 site
T G C G C A A C G C G T
FspAI  (2196)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (2221)
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
764 .. 1846  =  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
764 .. 1846  =  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
3986 .. 4801  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
3986 .. 4801  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
3276 .. 3864  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3276 .. 3864  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4894 .. 5349  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4894 .. 5349  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
2655 .. 2846  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2655 .. 2846  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
lacI promoter
686 .. 763  =  78 bp
lacI promoter
686 .. 763  =  78 bp
MCS
158 .. 225  =  68 bp
multiple cloning site
MCS
158 .. 225  =  68 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
lac operator
334 .. 358  =  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
334 .. 358  =  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).
T7 promoter
359 .. 377  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
359 .. 377  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
298 .. 303  =  6 bp
ribosome binding site
RBS
298 .. 303  =  6 bp
ribosome binding site
pelB signal sequence
224 .. 289  =  66 bp
22 amino acids  =  2.2 kDa
Product: leader peptide for secretion
pelB signal sequence
224 .. 289  =  66 bp
22 amino acids  =  2.2 kDa
Product: leader peptide for secretion
ORF:  514 .. 1014  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  1603 .. 1866  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  290 .. 517  =  228 bp
ORF:  75 amino acids  =  7.9 kDa
ORF:  887 .. 1846  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  3986 .. 4801  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  1902 .. 2258  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  2622 .. 2846  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  462 .. 725  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  1629 .. 1880  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  2255 .. 2623  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  349 .. 588  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
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