pET-50b(+)

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

Sequence Author: MilliporeSigma (Novagen)

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: pET & Duet Vectors (Novagen) | More Plasmid Sets
No matches
S-Tag PaeR7I - XhoI (147) PacI (141) AvrII (131) DraIII (6491) PsiI (6363) AlwNI (5004) PciI (4588) BspQI - SapI (4472) BstZ17I (4359) PflFI - Tth111I (4333) thrombin site Eco53kI (236) SacI (238) EagI - NotI (242) HindIII (249) SalI (255) PshAI (262) AscI (270) BsrGI (283) EcoRI (289) BamHI (295) Acc65I (302) KpnI - TspMI - XmaI (306) SmaI (308) KflI (310) HRV 3C site SacII (338) 6xHis SpeI (369) BglII (484) MscI (616) BfuAI - BspMI (818) NcoI (944) 6xHis ATG NdeI (1894) RBS XbaI (1932) T7 promoter SphI (2199) MluI (2724) BstEII (2905) NmeAIII (2930) PspOMI (2931) ApaI (2935) HpaI (3230) pET-50b(+) 6733 bp
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
DraIII  (6491)
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  (6363)
1 site
T T A T A A A A T A T T
AlwNI  (5004)
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.
PciI  (4588)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4472)
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  (4472)
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  (4359)
1 site
G T A T A C C A T A T G
PflFI  (4333)
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  (4333)
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.
AscI  (270)
1 site
G G C G C G C C C C G C G C G G
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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  (369)
1 site
A C T A G T T G A T C A
BglII  (484)
1 site
A G A T C T T C T A G A
MscI  (616)
1 site
T G G C C A A C C G G T
BfuAI  (818)
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  (818)
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.
NcoI  (944)
1 site
C C A T G G G G T A C C
NdeI  (1894)
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  (1932)
1 site
T C T A G A A G A T C T
SphI  (2199)
1 site
G C A T G C C G T A C G
MluI  (2724)
1 site
A C G C G T T G C G C A
BstEII  (2905)
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  (2930)
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  (2931)
1 site
G G G C C C C C C G G G
ApaI  (2935)
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  (3230)
1 site
G T T A A C C A A T T G
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
thrombin site
213 .. 230  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
213 .. 230  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
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
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
NusA
375 .. 1859  =  1485 bp
495 amino acids  =  54.9 kDa
Product: transcription elongation protein (N utilization substance protein A)
highly soluble in E. coli
NusA
375 .. 1859  =  1485 bp
495 amino acids  =  54.9 kDa
Product: transcription elongation protein (N utilization substance protein A)
highly soluble in E. coli
6xHis
1866 .. 1883  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1866 .. 1883  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
1893 .. 1895  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1893 .. 1895  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
lacI
2374 .. 3456  =  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
2374 .. 3456  =  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
5359 .. 6174  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
5359 .. 6174  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
4649 .. 5237  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4649 .. 5237  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6267 .. 6722  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6267 .. 6722  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
4028 .. 4219  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
4028 .. 4219  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
lacI promoter
2296 .. 2373  =  78 bp
lacI promoter
2296 .. 2373  =  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
lac operator
1940 .. 1964  =  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
1940 .. 1964  =  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
1965 .. 1983  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1965 .. 1983  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
1904 .. 1909  =  6 bp
ribosome binding site
RBS
1904 .. 1909  =  6 bp
ribosome binding site
ORF:  469 .. 705  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2497 .. 3456  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  5359 .. 6174  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  3995 .. 4219  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  345 .. 1673  =  1329 bp
ORF:  442 amino acids  =  51.3 kDa
ORF:  1866 .. 2624  =  759 bp
ORF:  252 amino acids  =  26.5 kDa
ORF:  3213 .. 3476  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2072 .. 2335  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  3239 .. 3490  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  3628 .. 3996  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  141 .. 1895  =  1755 bp
ORF:  584 amino acids  =  64.3 kDa
ORF:  1947 .. 2198  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
Click here to try SnapGene

Download pET-50b(+).dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

The maps, notes, and annotations in the zip file on this page 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.

Discover the most user-friendly molecular biology experience.