pENTR SD D-TOPO

Directional TOPO® cloning vector, for creating a Gateway® entry vector that is optimal for recombination with a prokaryotic destination vector.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTOPO Cloning VectorspENTR SD D-TOPO
PciI (1886) DrdI (1784) BssSI - BssSαI (1713) BciVI (1688) BsiHKAI (1576) ApaLI (1572) AlwNI (1477) AcuI - Eco57MI (1344) PflMI (974) Bpu10I (728) BsmBI - Esp3I (727) AsiSI - PvuI (711) BsrFI (665) SspI (636) NspI (1890) BspQI - SapI (2003) rrnB T2 terminator BsaHI (2219) AclI (2256) BbsI (2329) HincII - HpaI (2393) AhdI (2441) AflII (2446) AvaI - BsoBI (2452) PspOMI (2455) EcoO109I (2456) ApaI (2459) BtgI (2561) EagI - NotI - SacII (2564) AscI - BssHII (9) EcoRV (129) M13 rev NruI (368) EcoNI (623) pENTR™/SD/D-TOPO® 2597 bp
PciI  (1886)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (1784)
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  (1713)
1 site		 C A C G A G G T G C T C
BssSαI  (1713)
1 site		 C A C G A G G T G C T C
BciVI  (1688)
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.
BsiHKAI  (1576)
1 site		 G W G C W C C W C G W G
Sticky ends from different BsiHKAI sites may not be compatible.
ApaLI  (1572)
1 site		 G T G C A C C A C G T G
AlwNI  (1477)
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  (1344)
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  (1344)
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  (974)
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  (728)
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  (727)
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  (727)
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  (711)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (711)
1 site		 C G A T C G G C T A G C
BsrFI  (665)
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  (636)
1 site		 A A T A T T T T A T A A
NspI  (1890)
1 site		 R C A T G Y Y G T A C R
BspQI  (2003)
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  (2003)
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.
BsaHI  (2219)
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  (2256)
1 site		 A A C G T T T T G C A A
BbsI  (2329)
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.
HincII  (2393)
1 site		 G T Y R A C C A R Y T G
HpaI  (2393)
1 site		 G T T A A C C A A T T G
AhdI  (2441)
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  (2446)
1 site		 C T T A A G G A A T T C
AvaI  (2452)
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  (2452)
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.
PspOMI  (2455)
1 site		 G G G C C C C C C G G G
EcoO109I  (2456)
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.
ApaI  (2459)
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  (2561)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
EagI  (2564)
1 site		 C G G C C G G C C G G C
NotI  (2564)
1 site		 G C G G C C G C C G C C G G C G
SacII  (2564)
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.
AscI  (9)
1 site		 G G C G C G C C C C G C G C G G
BssHII  (9)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
EcoRV  (129)
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  (368)
1 site		 T C G C G A A G C G C T
EcoNI  (623)
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.
KanR
286 .. 1095  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
286 .. 1095  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
1242 .. 1830  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1242 .. 1830  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
attL2
17 .. 116  =  100 bp
recombination site for the Gateway® LR reaction
attL2
17 .. 116  =  100 bp
recombination site for the Gateway® LR reaction
attL1
2461 .. 2560  =  100 bp
recombination site for the Gateway® LR reaction
attL1
2461 .. 2560  =  100 bp
recombination site for the Gateway® LR reaction
rrnB T1 terminator
2279 .. 2365  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
2279 .. 2365  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T2 terminator
2160 .. 2187  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
2160 .. 2187  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
RBS
2572 .. 2592  =  21 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
2572 .. 2592  =  21 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
134 .. 152  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
134 .. 152  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
157 .. 173  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
157 .. 173  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2429 .. 2445  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2429 .. 2445  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  286 .. 1095  =  810 bp
ORF:  269 amino acids  =  30.8 kDa
ORF:  2286 .. 2600  =  315 bp
ORF:  104 amino acids  =  11.5 kDa  (no start codon)
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Download pENTR SD D-TOPO.dna file

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Individual Sequences & Maps

pBAD-TOPO (linearized)
pcDNA5 FRT TO-TOPO (linearized)
pEF6 V5-His-TOPO (linearized)
pGlow-TOPO (linearized)
pBAD Thio-TOPO (linearized)
pcDNA5 FRT V5-His-TOPO (linearized)
pENTR D-TOPO
pIB V5-His-TOPO (linearized)
pBAD202 D-TOPO
pcDNA6.2 C-EmGFP-GW TOPO (linearized)
pENTR SD D-TOPO
pLenti6.3 V5-TOPO (linearized)
pBlue-TOPO (linearized)
pcDNA6.2 C-YFP-GW TOPO (linearized)
pENTR TEV D-TOPO
pLenti6 V5-D-TOPO
pcDNA3.1D V5-His-TOPO
pcDNA6.2 N-EmGFP-GW TOPO (linearized)
pENTR5'-TOPO (linearized)
pLenti7.3 V5-TOPO (linearized)
pcDNA3.2 V5 GW D-TOPO
pCR-XL-2-TOPO (linearized)
pET100 D-TOPO
pLPS-B (linearized)
pcDNA6.2 cGeneBLAzer-GW D-TOPO
pCR-XL-TOPO (linearized)
pET101 D-TOPO
pMT V5-His-TOPO (linearized)
pcDNA6.2 V5 GW D-TOPO
pCR2.1-TOPO (linearized)
pET102 D-TOPO
pOptiVEC-TOPO (linearized)
pcDNA3.1 CT-GFP-TOPO (linearized)
pCR4-TOPO (linearized)
pET151 D-TOPO
pSecTag FRT V5-His-TOPO (linearized)
pcDNA3.1 NT-GFP-TOPO (linearized)
pCR4Blunt-TOPO (linearized)
pET160 GW D-TOPO
pTOP Blunt V2 (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pCR8 GW TOPO (linearized)
pET161 GW D-TOPO
pTrcHis-TOPO (linearized)
pcDNA3.3-TOPO (linearized)
pCR-Blunt II-TOPO (linearized)
pET200 D-TOPO
pTrcHis2-TOPO (linearized)
pcDNA3.4-TOPO (linearized)
pCRII-TOPO (linearized)
pEXP5-CT TOPO (linearized)
pYES2.1 V5-His-TOPO (linearized)
pcDNA4 HisMax-TOPO (linearized)
pEF5 FRT V5-D-TOPO
pEXP5-NT 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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