pBlue-TOPO (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products to measure promoter activity using β-galactosidase.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTA and GC Cloning VectorspBlue-TOPO (linearized)
T7 promoter BglII (7689) SgrDI (7675) ScaI (7234) AhdI (6754) BsmI (5430) BstBI (5192) RsrII (5026) PflFI - Tth111I (4628) MscI (4592) PluTI (4513) SfoI (4511) NarI (4510) KasI (4509) SmaI (4323) TspMI - XmaI (4321) AvrII (4300) StuI (4299) BseRI (4296) CsiI - SexAI * (4067) AflII (7739) BamHI (7760) SpeI (7766) End (7793) Start (1) 6xHis Xpress™ tag Bpu10I (642) BspDI - ClaI (961) EcoRV (1250) MauBI (1635) AfeI (1972) BsiWI (2908) NdeI (3094) BlpI (3150) NotI (3224) PaeR7I - PspXI - XhoI (3230) XbaI (3236) EcoO109I - PspOMI (3242) ApaI (3246) pBlue-TOPO® 7792 bp
BglII  (7689)
1 site		 A G A T C T T C T A G A
SgrDI  (7675)
1 site		 C G T C G A C G G C A G C T G C
ScaI  (7234)
1 site		 A G T A C T T C A T G A
AhdI  (6754)
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.
BsmI  (5430)
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.
BstBI  (5192)
1 site		 T T C G A A A A G C T T
RsrII  (5026)
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.
PflFI  (4628)
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  (4628)
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.
MscI  (4592)
1 site		 T G G C C A A C C G G T
PluTI  (4513)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (4511)
1 site		 G G C G C C C C G C G G
NarI  (4510)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (4509)
1 site		 G G C G C C C C G C G G
SmaI  (4323)
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  (4321)
1 site		 C C C G G G G G G C C C
XmaI  (4321)
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.
AvrII  (4300)
1 site		 C C T A G G G G A T C C
StuI  (4299)
1 site		 A G G C C T T C C G G A
BseRI  (4296)
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.
CsiI  (4067)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (4067)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AflII  (7739)
1 site		 C T T A A G G A A T T C
BamHI  (7760)
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.
SpeI  (7766)
1 site		 A C T A G T T G A T C A
End  (7793)
0 sites
Start  (1)
0 sites
Bpu10I  (642)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BspDI  (961)
1 site		 A T C G A T T A G C T A
ClaI  (961)
1 site		 A T C G A T T A G C T A
EcoRV  (1250)
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.
MauBI  (1635)
1 site		 C G C G C G C G G C G C G C G C
AfeI  (1972)
1 site		 A G C G C T T C G C G A
BsiWI  (2908)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
NdeI  (3094)
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.
BlpI  (3150)
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.
NotI  (3224)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (3230)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3230)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (3230)
1 site		 C T C G A G G A G C T C
XbaI  (3236)
1 site		 T C T A G A A G A T C T
EcoO109I  (3242)
1 site		 R G G N C C Y Y C C N G G R
Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (3242)
1 site		 G G G C C C C C C G G G
ApaI  (3246)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
ATG
27 .. 29  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
27 .. 29  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
39 .. 56  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
39 .. 56  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 tag
60 .. 92  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 tag
60 .. 92  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
Xpress™ tag
96 .. 119  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 1:  
   96 .. 104  =  9 bp
   3 amino acids  =  409.4 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
96 .. 119  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 2:  enterokinase site  
   105 .. 119  =  15 bp
   5 amino acids  =  606.5 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
96 .. 119  =  24 bp
8 amino acids  =  998.0 Da
2 segments
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
lacZ
150 .. 3197  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
150 .. 3197  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
AmpR
6681 .. 7541  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6681 .. 7472  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6681 .. 7541  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7473 .. 7541  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6681 .. 7541  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
4382 .. 5176  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
4382 .. 5176  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
5922 .. 6510  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
5922 .. 6510  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
f1 ori
3543 .. 3971  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3543 .. 3971  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
3985 .. 4315  =  331 bp
SV40 enhancer and early promoter
SV40 promoter
3985 .. 4315  =  331 bp
SV40 enhancer and early promoter
bGH poly(A) signal
3273 .. 3497  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
3273 .. 3497  =  225 bp
bovine growth hormone polyadenylation signal
SV40 poly(A) signal
5350 .. 5471  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5350 .. 5471  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7542 .. 7646  =  105 bp
AmpR promoter
7542 .. 7646  =  105 bp
T7 promoter
7694 .. 7712  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
7694 .. 7712  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
4166 .. 4301  =  136 bp
SV40 origin of replication
SV40 ori
4166 .. 4301  =  136 bp
SV40 origin of replication
ORF:  6811 .. 7077  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3494 .. 3760  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  4382 .. 5176  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3 .. 3197  =  3195 bp
ORF:  1064 amino acids  =  120.9 kDa  (no start codon)
ORF:  4554 .. 4940  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  996 .. 1310  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2421 .. 2657  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  6681 .. 7541  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2060 .. 2371  =  312 bp
ORF:  103 amino acids  =  10.4 kDa
ORF:  4691 .. 5227  =  537 bp
ORF:  178 amino acids  =  19.8 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)
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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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