Resources
Plasmid Files

pET-48b(+)

Bacterial vector for expressing proteins with a cleavable N-terminal thioredoxin tag and a cleavable C-terminal S-tag.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pET-48b(+).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
Download Free Trial Get SnapGene Viewer

 BstBI (187) PaeR7I - XhoI (147) PacI (141) AvrII (131) BlpI (80) T7 terminator DraIII (5363) PsiI (5235) NruI (4319) AcuI (4008) AlwNI (3876) BssS α I (3633) PciI (3460) BspQI - SapI (3344) BstZ17I (3231) PflFI - Tth111I (3205) S-Tag thrombin site Eco53kI (236) SacI (238) EagI - NotI (242) HindIII (249) SalI (255) PshAI (262) ZraI (264) AatII (266) AscI (270) StuI (280) BsrGI (283) EcoRI (289) BamHI (295) Acc65I (302) KpnI - TspMI - XmaI (306) SmaI (308) KflI (310) HRV 3C site SacII (338) 6xHis SpeI (417) MscI (426) RsrII (664) NdeI (766) RBS XbaI (804) T7 promoter SgrAI (915) SphI (1071) BstAPI (1279) MluI (1596) BclI * (1610) BstEII (1777) NmeAIII (1802) PspOMI (1803) ApaI (1807) HpaI (2102) pET-48b(+) 5605 bp
BstBI  (187)
1 site
T T C G A A A A G C T T
PaeR7I  (147)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (147)
1 site
C T C G A G G A G C T C
PacI  (141)
1 site
T T A A T T A A A A T T A A T T
AvrII  (131)
1 site
C C T A G G G G A T C C
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.
DraIII  (5363)
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  (5235)
1 site
T T A T A A A A T A T T
NruI  (4319)
1 site
T C G C G A A G C G C T
AcuI  (4008)
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  (3876)
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  (3633)
1 site
C A C G A G G T G C T C
PciI  (3460)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3344)
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  (3344)
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  (3231)
1 site
G T A T A C C A T A T G
PflFI  (3205)
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  (3205)
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.
Eco53kI  (236)
1 site
G A G C T C C T C G A G
SacI  (238)
1 site
G A G C T C C T C G A G
EagI  (242)
1 site
C G G C C G G C C G G C
NotI  (242)
1 site
G C G G C C G C C G C C G G C G
HindIII  (249)
1 site
A A G C T T T T C G A A
SalI  (255)
1 site
G T C G A C C A G C T G
PshAI  (262)
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.
ZraI  (264)
1 site
G A C G T C C T G C A G
AatII  (266)
1 site
G A C G T C C T G C A G
AscI  (270)
1 site
G G C G C G C C C C G C G C G G
StuI  (280)
1 site
A G G C C T T C C G G A
BsrGI  (283)
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.
EcoRI  (289)
1 site
G A A T T C C T T A A G
BamHI  (295)
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.
Acc65I  (302)
1 site
G G T A C C C C A T G G
KpnI  (306)
1 site
G G T A C C C C A T G G
TspMI  (306)
1 site
C C C G G G G G G C C C
XmaI  (306)
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.
SmaI  (308)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KflI  (310)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
SacII  (338)
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  (417)
1 site
A C T A G T T G A T C A
MscI  (426)
1 site
T G G C C A A C C G G T
RsrII  (664)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
NdeI  (766)
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  (804)
1 site
T C T A G A A G A T C T
SgrAI  (915)
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  (1071)
1 site
G C A T G C C G T A C G
BstAPI  (1279)
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  (1596)
1 site
A C G C G T T G C G C A
BclI  (1610)
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  (1777)
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  (1802)
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  (1803)
1 site
G G G C C C C C C G G G
ApaI  (1807)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
HpaI  (2102)
1 site
G T T A A C C A A T T G
lacI
1246 .. 2328  =  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
1246 .. 2328  =  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
4231 .. 5046  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
4231 .. 5046  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
3521 .. 4109  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3521 .. 4109  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
5139 .. 5594  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
5139 .. 5594  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
TrxA
441 .. 767  =  327 bp
109 amino acids  =  11.8 kDa
Product: E. coli thioredoxin
TrxA
441 .. 767  =  327 bp
109 amino acids  =  11.8 kDa
Product: E. coli thioredoxin
rop
2900 .. 3091  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2900 .. 3091  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
lacI promoter
1168 .. 1245  =  78 bp
lacI promoter
1168 .. 1245  =  78 bp
MCS
234 .. 308  =  75 bp
multiple cloning site
MCS
234 .. 308  =  75 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
168 .. 212  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
S-Tag
168 .. 212  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
lac operator
812 .. 836  =  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
812 .. 836  =  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).
HRV 3C site
309 .. 332  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human
rhinovirus 3C protease
HRV 3C site
309 .. 332  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human
rhinovirus 3C protease
T7 promoter
837 .. 855  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
837 .. 855  =  19 bp
promoter for bacteriophage T7 RNA polymerase
thrombin site
213 .. 230  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
213 .. 230  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
6xHis
342 .. 359  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
342 .. 359  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
776 .. 781  =  6 bp
ribosome binding site
RBS
776 .. 781  =  6 bp
ribosome binding site
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2017 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter