Resources
Plasmid Files

pBAD/Thio-TOPO® (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning and arabinose-inducible expression of a His-Patch thioredoxin fusion protein.

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

pBAD Thio-TOPO (linearized) Sequence and MappBAD Thio-TOPO (linearized).dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
Download Free Trial Get SnapGene Viewer

 PflMI (3660) BspEI (3455) NruI (3448) EcoRV (3126) AflII (3041) BsaBI (2981) NsiI (2749) SphI (2747) MauBI (2725) BsmBI (2659) PfoI (2657) PflFI - Tth111I (2558) BsaAI (2552) BstZ17I (2533) AccI (2532) NdeI (2481) PciI (2302) AlwNI (1893) BanI (1461) AhdI (1414) BstEII (3817) BsmI (3837) NcoI - StyI (4084) AflII (4140) AleI (4181) RsrII (4188) BspDI * - ClaI * (4270) TspMI - XmaI (4280) SmaI (4282) NgoMIV (4413) NaeI (4415) End (4454) Start (1) BseRI (28) V5 tag 6xHis PmeI (91) HincII (872) ScaI (933) PvuI (1045) FspI (1191) AseI (1239) NmeAIII (1267) BglI (1296) pBAD/Thio-TOPO® 4453 bp
PflMI  (3660)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BspEI  (3455)
1 site
T C C G G A A G G C C T
NruI  (3448)
1 site
T C G C G A A G C G C T
EcoRV  (3126)
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.
AflII  (3041)
2 sites
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
BsaBI  (2981)
1 site
G A T N N N N A T C C T A N N N N T A G
NsiI  (2749)
1 site
A T G C A T T A C G T A
SphI  (2747)
1 site
G C A T G C C G T A C G
MauBI  (2725)
1 site
C G C G C G C G G C G C G C G C
BsmBI  (2659)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PfoI  (2657)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
PflFI  (2558)
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  (2558)
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.
BsaAI  (2552)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2533)
1 site
G T A T A C C A T A T G
AccI  (2532)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
NdeI  (2481)
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.
PciI  (2302)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (1893)
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.
BanI  (1461)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
AhdI  (1414)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BstEII  (3817)
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.
BsmI  (3837)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
NcoI  (4084)
1 site
C C A T G G G G T A C C
StyI  (4084)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
AflII  (4140)
2 sites
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
AleI  (4181)
1 site
C A C N N N N G T G G T G N N N N C A C
RsrII  (4188)
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.
BspDI  (4270)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4270)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
TspMI  (4280)
1 site
C C C G G G G G G C C C
XmaI  (4280)
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  (4282)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NgoMIV  (4413)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (4415)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
End  (4454)
0 sites
Start  (1)
0 sites
BseRI  (28)
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.
PmeI  (91)
1 site
G T T T A A A C C A A A T T T G
HincII  (872)
1 site
G T Y R A C C A R Y T G
ScaI  (933)
1 site
A G T A C T T C A T G A
PvuI  (1045)
1 site
C G A T C G G C T A G C
FspI  (1191)
1 site
T G C G C A A C G C G T
AseI  (1239)
1 site
A T T A A T T A A T T A
NmeAIII  (1267)
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).
BglI  (1296)
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.
araC
2836 .. 3714  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
araC
2836 .. 3714  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
AmpR
627 .. 1487  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   627 .. 695  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
627 .. 1487  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   696 .. 1487  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
627 .. 1487  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1658 .. 2246  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
1658 .. 2246  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
HP-thioredoxin
4086 .. 4415  =  330 bp
110 amino acids  =  11.9 kDa
Product: His-Patch thioredoxin
The histidines at positions 8, 32, and 64 coordinate
a divalent cation, allowing HP-thioredoxin to be
purified on metal-chelating resins.
HP-thioredoxin
4086 .. 4415  =  330 bp
110 amino acids  =  11.9 kDa
Product: His-Patch thioredoxin
The histidines at positions 8, 32, and 64 coordinate
a divalent cation, allowing HP-thioredoxin to be
purified on metal-chelating resins.
araBAD promoter
3860 .. 4025  =  166 bp
promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction
araBAD promoter
3860 .. 4025  =  166 bp
promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction
AmpR promoter
535 .. 626  =  92 bp
AmpR promoter
535 .. 626  =  92 bp
rrnB T1 terminator
310 .. 396  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
310 .. 396  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
V5 tag
16 .. 57  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
16 .. 57  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
rrnB T2 terminator
488 .. 515  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
488 .. 515  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
6xHis
67 .. 84  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
67 .. 84  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
enterokinase site
4431 .. 4445  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
4431 .. 4445  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
RBS
4069 .. 4074  =  6 bp
ribosome binding site
RBS
4069 .. 4074  =  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 © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter