pENTR5'-TOPO (linearized)

Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products to create a multisite Gateway® entry clone.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTA and GC Cloning VectorspENTR5'-TOPO (linearized)
PciI (1969) DrdI (1867) BssSI - BssSαI (1796) BciVI (1771) PspFI (1669) BseYI (1665) ApaLI (1655) AlwNI (1560) AcuI - Eco57MI (1427) PflMI (1057) Bpu10I (811) BsmBI - Esp3I (810) AsiSI - PvuI (794) BsrFI (748) SspI (719) EcoNI (706) NspI (1973) BspQI - SapI (2086) PvuII (2149) rrnB T2 terminator BsaHI (2302) AclI (2339) BbsI (2412) AhdI (2524) AflII (2529) PspOMI (2538) ApaI (2542) BtgI - NcoI - StyI (2556) PsiI (2636) End (2680) Start (1) BsrGI - TatI (23) attR1 NdeI (125) XmnI (178) XbaI (188) PstI (199) EagI - NotI (203) EcoRV (212) NruI (451) pENTR™5'-TOPO® 2679 bp
PciI  (1969)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (1867)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
BssSI  (1796)
1 site		 C A C G A G G T G C T C
BssSαI  (1796)
1 site		 C A C G A G G T G C T C
BciVI  (1771)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
PspFI  (1669)
1 site		 C C C A G C G G G T C G
BseYI  (1665)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
ApaLI  (1655)
1 site		 G T G C A C C A C G T G
AlwNI  (1560)
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.
AcuI  (1427)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
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).
Eco57MI  (1427)
1 site		 C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14
Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PflMI  (1057)
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.
Bpu10I  (811)
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.
BsmBI  (810)
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  (810)
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.
AsiSI  (794)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (794)
1 site		 C G A T C G G C T A G C
BsrFI  (748)
1 site		 R C C G G Y Y G G C C R
Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
SspI  (719)
1 site		 A A T A T T T T A T A A
EcoNI  (706)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
NspI  (1973)
1 site		 R C A T G Y Y G T A C R
BspQI  (2086)
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  (2086)
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.
PvuII  (2149)
1 site		 C A G C T G G T C G A C
BsaHI  (2302)
1 site		 G R C G Y C C Y G C R G
BsaHI is typically used at 37°C, but is even more active at 60°C.
AclI  (2339)
1 site		 A A C G T T T T G C A A
BbsI  (2412)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
AhdI  (2524)
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.
AflII  (2529)
1 site		 C T T A A G G A A T T C
PspOMI  (2538)
1 site		 G G G C C C C C C G G G
ApaI  (2542)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BtgI  (2556)
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  (2556)
1 site		 C C A T G G G G T A C C
StyI  (2556)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
PsiI  (2636)
1 site		 T T A T A A A A T A T T
End  (2680)
0 sites
Start  (1)
0 sites
BsrGI  (23)
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.
TatI  (23)
1 site		 W G T A C W W C A T G W
NdeI  (125)
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.
XmnI  (178)
1 site		 G A A N N N N T T C C T T N N N N A A G
XbaI  (188)
1 site		 T C T A G A A G A T C T
PstI  (199)
1 site		 C T G C A G G A C G T C
EagI  (203)
1 site		 C G G C C G G C C G G C
NotI  (203)
1 site		 G C G G C C G C C G C C G G C G
EcoRV  (212)
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.
NruI  (451)
1 site		 T C G C G A A G C G C T
KanR
369 .. 1178  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
369 .. 1178  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
1325 .. 1913  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1325 .. 1913  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
attR1
17 .. 140  =  124 bp
recombination site for the Gateway® LR reaction
attR1
17 .. 140  =  124 bp
recombination site for the Gateway® LR reaction
attL4
2568 .. 2663  =  96 bp
recombination site for the Gateway® LR reaction
attL4
2568 .. 2663  =  96 bp
recombination site for the Gateway® LR reaction
rrnB T1 terminator
2362 .. 2448  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
2362 .. 2448  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T2 terminator
2243 .. 2270  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
2243 .. 2270  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
T7 promoter
217 .. 235  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
217 .. 235  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
240 .. 256  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
240 .. 256  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2512 .. 2528  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2512 .. 2528  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  369 .. 1178  =  810 bp
ORF:  269 amino acids  =  30.8 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)
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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