pLenti6 V5-D-TOPO

Lentiviral directional TOPO® cloning vector for C-terminal tagging of a protein with a V5 epitope tag.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTOPO Cloning VectorspLenti6 V5-D-TOPO
PciI (4114) AhdI (3226) ScaI (2745) DraIII (2086) NaeI (1980) NgoMIV (1978) SfiI (1598) KpnI (978) Acc65I (974) BlpI (955) KasI (4937) NarI (4938) SfoI (4939) PluTI (4941) AccI (5262) NotI (5444) EcoNI (5466) MfeI (5485) BbvCI - Bpu10I (5720) AleI (5873) KflI (6230) BspDI - ClaI (6319) NdeI (6513) SnaBI (6619) BsmBI - Esp3I (6868) BamHI (6924) SpeI (6930) BstXI (6943) PaeR7I - PspXI - XhoI (6) PspOMI (18) ApaI (22) SacII (25) BstBI (29) MluI (73) AgeI (79) CsiI - SexAI * (198) TspMI - XmaI (452) SmaI (454) BclI * (482) PmlI (491) HincII (496) MscI (563) BsaBI * (858) pLenti6/V5-D-TOPO® 6959 bp
PciI  (4114)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (3226)
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.
ScaI  (2745)
1 site		 A G T A C T T C A T G A
DraIII  (2086)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
NaeI  (1980)
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.
NgoMIV  (1978)
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.
SfiI  (1598)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
KpnI  (978)
1 site		 G G T A C C C C A T G G
Acc65I  (974)
1 site		 G G T A C C C C A T G G
BlpI  (955)
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.
KasI  (4937)
1 site		 G G C G C C C C G C G G
NarI  (4938)
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.
SfoI  (4939)
1 site		 G G C G C C C C G C G G
PluTI  (4941)
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.
AccI  (5262)
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.
NotI  (5444)
1 site		 G C G G C C G C C G C C G G C G
EcoNI  (5466)
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.
MfeI  (5485)
1 site		 C A A T T G G T T A A C
BbvCI  (5720)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (5720)
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.
AleI  (5873)
1 site		 C A C N N N N G T G G T G N N N N C A C
KflI  (6230)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
BspDI  (6319)
1 site		 A T C G A T T A G C T A
ClaI  (6319)
1 site		 A T C G A T T A G C T A
NdeI  (6513)
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.
SnaBI  (6619)
1 site		 T A C G T A A T G C A T
BsmBI  (6868)
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  (6868)
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.
BamHI  (6924)
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  (6930)
1 site		 A C T A G T T G A T C A
BstXI  (6943)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
PaeR7I  (6)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (6)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (6)
1 site		 C T C G A G G A G C T C
PspOMI  (18)
1 site		 G G G C C C C C C G G G
ApaI  (22)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
SacII  (25)
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.
BstBI  (29)
1 site		 T T C G A A A A G C T T
MluI  (73)
1 site		 A C G C G T T G C G C A
AgeI  (79)
1 site		 A C C G G T T G G C C A
CsiI  (198)
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  (198)
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.
TspMI  (452)
1 site		 C C C G G G G G G C C C
XmaI  (452)
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.
SmaI  (454)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BclI  (482)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
PmlI  (491)
1 site		 C A C G T G G T G C A C
HincII  (496)
1 site		 G T Y R A C C A R Y T G
MscI  (563)
1 site		 T G G C C A A C C G G T
BsaBI  (858)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
AmpR
2439 .. 3299  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2439 .. 2507  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2439 .. 3299  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2508 .. 3299  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2439 .. 3299  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3470 .. 4058  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3470 .. 4058  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1853 .. 2308  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1853 .. 2308  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
560 .. 958  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
560 .. 958  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
117 .. 446  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
117 .. 446  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
6340 .. 6643  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
6340 .. 6643  =  304 bp
human cytomegalovirus immediate early enhancer
3' LTR (ΔU3)
1045 .. 1278  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
1045 .. 1278  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RRE
5599 .. 5832  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
5599 .. 5832  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RSV promoter
4527 .. 4753  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
4527 .. 4753  =  227 bp
Rous sarcoma virus enhancer/promoter
CMV promoter
6644 .. 6847  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
6644 .. 6847  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR (truncated)
4754 .. 4934  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
4754 .. 4934  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
4981 .. 5106  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
4981 .. 5106  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
1350 .. 1471  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1350 .. 1471  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2334 .. 2438  =  105 bp
AmpR promoter
2334 .. 2438  =  105 bp
EM7 promoter
494 .. 541  =  48 bp
synthetic bacterial promoter
EM7 promoter
494 .. 541  =  48 bp
synthetic bacterial promoter
V5 tag
34 .. 75  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
34 .. 75  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
SV40 ori
297 .. 432  =  136 bp
SV40 origin of replication
SV40 ori
297 .. 432  =  136 bp
SV40 origin of replication
ORF:  1372 .. 1608  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  4456 .. 4701  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  560 .. 958  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  5477 .. 6325  =  849 bp
ORF:  282 amino acids  =  32.5 kDa
ORF:  894 .. 1124  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  2439 .. 3299  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  5392 .. 5772  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  4383 .. 4682  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  6726 .. 6962  =  237 bp
ORF:  78 amino acids  =  8.5 kDa  (no start codon)
ORF:  506 .. 862  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  2903 .. 3169  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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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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