pBAD-TOPO (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning and arabinose-inducible expression.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTA and GC Cloning VectorspBAD-TOPO (linearized)
PflMI (3660) BspEI (3455) NruI (3448) EcoRV (3126) AflII (3041) BsaBI (2981) AvaI - BsoBI (2894) NsiI (2749) SphI (2747) MauBI (2725) BsmBI - Esp3I (2659) PfoI (2657) PflFI - Tth111I (2558) BsaAI (2552) BstZ17I (2533) AccI (2532) NdeI (2481) PciI (2302) AlwNI (1893) BanI (1461) BstEII (3817) BsmI (3837) BtgI - NcoI - StyI (4084) BanII (4092) End (4127) Start (1) BseRI (28) V5 tag 6xHis PmeI (91) HincII (872) ScaI (933) PvuI (1045) FspI (1191) AseI (1239) NmeAIII (1267) BglI (1296) AhdI (1414) pBAD-TOPO® 4126 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)
1 site		 C T T A A G G A A T T C
BsaBI  (2981)
1 site		 G A T N N N N A T C C T A N N N N T A G
AvaI  (2894)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2894)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2659)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I 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.
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.
BtgI  (4084)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI 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.
BanII  (4092)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
End  (4127)
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.
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.
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
2 segments
   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
2 segments
   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
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
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
ATG
4086 .. 4088  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4086 .. 4088  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
enterokinase site
4104 .. 4118  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
4104 .. 4118  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
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
RBS
4069 .. 4072  =  4 bp
ribosome binding site
RBS
4069 .. 4072  =  4 bp
ribosome binding site
ORF:  3400 .. 3648  =  249 bp
ORF:  82 amino acids  =  9.3 kDa
ORF:  506 .. 730  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  627 .. 1487  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1091 .. 1357  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  151 .. 477  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  2836 .. 3765  =  930 bp
ORF:  309 amino acids  =  35.2 kDa
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Individual Sequences & Maps

Allele TA Vector (linearized)
pCR2.1-TOPO (linearized)
pJAZZ-OK GC (left arm)
pSpark TA
pBAD-TOPO (linearized)
pCR4-TOPO (linearized)
pJAZZ-OK GC (right arm)
pSpark TA Done
pBAD Thio-TOPO (linearized)
pCR8 GW TOPO (linearized)
pLenti6.3 V5-TOPO (linearized)
pSTBlue-1 AccepTor
pBlue-TOPO (linearized)
pCRII-TOPO (linearized)
pLenti7.3 V5-TOPO (linearized)
pT7Blue T-Vector
pcDNA3.1 CT-GFP-TOPO (linearized)
pDrive (linearized)
pLPS-T (linearized)
pTA Plus (linearized)
pcDNA3.1 NT-GFP-TOPO (linearized)
pEF6 V5-His-TOPO (linearized)
pLUG (linearized)
pTargeT (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pENTR5'-TOPO (linearized)
pLUG-Multi (linearized)
pTOP TA V2 (linearized)
pcDNA3.3-TOPO (linearized)
pET SUMO (linearized)
pMT V5-His-TOPO (linearized)
pTrcHis-TOPO (linearized)
pcDNA3.4-TOPO (linearized)
pETBlue-1 AccepTor
pOptiVEC-TOPO (linearized)
pTrcHis2-TOPO (linearized)
pcDNA4 HisMax-TOPO (linearized)
pEXP5-CT TOPO (linearized)
pPrime (linearized)
pTZ57R_T
pcDNA5 FRT TO-TOPO (linearized)
pEXP5-NT TOPO (linearized)
pQE-30 UA (linearized)
pUC57-T
pcDNA5 FRT V5-His-TOPO (linearized)
pGC Blue (linearized)
pSC-A-amp kan (linearized)
pYES2.1 V5-His-TOPO (linearized)
pcDNA6.2 C-EmGFP-GW TOPO (linearized)
pGEM-T (linearized)
pSecTag FRT V5-His-TOPO (linearized)
RBC T&A Cloning Vector (linearized)
pcDNA6.2 C-YFP-GW TOPO (linearized)
pGEM-T Easy (linearized)
pSMART GC HK (linearized)
T-Vector pMD19 (Simple)
pcDNA6.2 N-EmGFP-GW TOPO (linearized)
pGlow-TOPO (linearized)
pSMART GC LK (linearized)
T-Vector pMD20
pCR-XL-TOPO (linearized)
pIB V5-His-TOPO (linearized)

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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