pLenti6.3 V5-TOPO (linearized)

Linearized lentiviral expression vector with a blasticidin resistance gene with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsTA and GC Cloning VectorspLenti6.3 V5-TOPO (linearized)
KasI (5543) PciI (4720) AhdI (3832) ScaI (3351) DraIII (2692) SfiI (2204) BlpI (1561) MscI (1169) PmlI (1097) NarI (5544) SfoI (5545) PluTI (5547) EcoNI (6072) MfeI (6091) BbvCI - Bpu10I (6326) AleI (6479) KflI (6835) HpaI (6929) BspDI - ClaI (7052) NdeI (7246) SnaBI (7352) BsmBI - Esp3I (7601) BamHI (7657) SpeI (7663) End (7691) Start (1) PstI (16) EcoRV (19) PaeR7I - PspXI - XhoI (40) PspOMI (52) ApaI (56) BstBI (63) MluI (107) PflMI (374) PfoI (406) CsiI - SexAI * (804) TspMI - XmaI (1058) SmaI (1060) BclI * (1088) pLenti6.3/V5-TOPO® 7690 bp
KasI  (5543)
1 site		 G G C G C C C C G C G G
PciI  (4720)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (3832)
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  (3351)
1 site		 A G T A C T T C A T G A
DraIII  (2692)
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.
SfiI  (2204)
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.
BlpI  (1561)
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.
MscI  (1169)
1 site		 T G G C C A A C C G G T
PmlI  (1097)
1 site		 C A C G T G G T G C A C
NarI  (5544)
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  (5545)
1 site		 G G C G C C C C G C G G
PluTI  (5547)
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.
EcoNI  (6072)
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  (6091)
1 site		 C A A T T G G T T A A C
BbvCI  (6326)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (6326)
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  (6479)
1 site		 C A C N N N N G T G G T G N N N N C A C
KflI  (6835)
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.
HpaI  (6929)
1 site		 G T T A A C C A A T T G
BspDI  (7052)
1 site		 A T C G A T T A G C T A
ClaI  (7052)
1 site		 A T C G A T T A G C T A
NdeI  (7246)
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  (7352)
1 site		 T A C G T A A T G C A T
BsmBI  (7601)
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  (7601)
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  (7657)
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  (7663)
1 site		 A C T A G T T G A T C A
End  (7691)
0 sites
Start  (1)
0 sites
PstI  (16)
1 site		 C T G C A G G A C G T C
EcoRV  (19)
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.
PaeR7I  (40)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (40)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (40)
1 site		 C T C G A G G A G C T C
PspOMI  (52)
1 site		 G G G C C C C C C G G G
ApaI  (56)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BstBI  (63)
1 site		 T T C G A A A A G C T T
MluI  (107)
1 site		 A C G C G T T G C G C A
PflMI  (374)
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.
PfoI  (406)
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.
CsiI  (804)
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  (804)
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  (1058)
1 site		 C C C G G G G G G C C C
XmaI  (1058)
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  (1060)
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  (1088)
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.
AmpR
3045 .. 3905  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3045 .. 3113  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3045 .. 3905  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3114 .. 3905  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3045 .. 3905  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
133 .. 721  =  589 bp
woodchuck posttranscriptional regulatory element
WPRE
133 .. 721  =  589 bp
woodchuck posttranscriptional regulatory element
ori
4076 .. 4664  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
4076 .. 4664  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
f1 ori
2459 .. 2914  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2459 .. 2914  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
1166 .. 1564  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
1166 .. 1564  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
736 .. 1052  =  317 bp
SV40 enhancer and early promoter
SV40 promoter
736 .. 1052  =  317 bp
SV40 enhancer and early promoter
CMV enhancer
7073 .. 7376  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
7073 .. 7376  =  304 bp
human cytomegalovirus immediate early enhancer
3'LTR (ΔU3)
1651 .. 1884  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3'LTR (ΔU3)
1651 .. 1884  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
HIV-1 RRE
6205 .. 6438  =  234 bp
The Rev response element (RRE) of human immunodeficiency virus type 1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
HIV-1 RRE
6205 .. 6438  =  234 bp
The Rev response element (RRE) of human immunodeficiency virus type 1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RSV promoter
5131 .. 5354  =  224 bp
Rous sarcoma virus promoter
RSV promoter
5131 .. 5354  =  224 bp
Rous sarcoma virus promoter
CMV promoter
7377 .. 7580  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
7377 .. 7580  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR
5360 .. 5540  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR
5360 .. 5540  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 ψ
5587 .. 5712  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 ψ
5587 .. 5712  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
1956 .. 2077  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1956 .. 2077  =  122 bp
SV40 polyadenylation signal
HIV-1 cPPT
6933 .. 7050  =  118 bp
central polypurine tract from pol gene of human immunodeficiency virus type 1
HIV-1 cPPT
6933 .. 7050  =  118 bp
central polypurine tract from pol gene of human immunodeficiency virus type 1
AmpR promoter
2940 .. 3044  =  105 bp
AmpR promoter
2940 .. 3044  =  105 bp
EM7 promoter
1100 .. 1147  =  48 bp
synthetic bacterial promoter
EM7 promoter
1100 .. 1147  =  48 bp
synthetic bacterial promoter
V5 tag
68 .. 109  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
68 .. 109  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
SV40 ori
903 .. 1038  =  136 bp
SV40 origin of replication
SV40 ori
903 .. 1038  =  136 bp
SV40 origin of replication
ORF:  124 .. 882  =  759 bp
ORF:  252 amino acids  =  27.2 kDa
ORF:  1978 .. 2214  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  5062 .. 5307  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  6544 .. 7092  =  549 bp
ORF:  182 amino acids  =  21.2 kDa
ORF:  236 .. 532  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  728 .. 1000  =  273 bp
ORF:  90 amino acids  =  10.3 kDa
ORF:  1166 .. 1564  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  6083 .. 6583  =  501 bp
ORF:  166 amino acids  =  18.4 kDa
ORF:  543 .. 797  =  255 bp
ORF:  84 amino acids  =  8.9 kDa
ORF:  1500 .. 1730  =  231 bp
ORF:  76 amino acids  =  8.6 kDa
ORF:  3045 .. 3905  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4989 .. 5288  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  1112 .. 1468  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  3509 .. 3775  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5998 .. 6378  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  7459 .. 7689  =  231 bp
ORF:  76 amino acids  =  8.2 kDa  (no start codon)
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Individual Sequences & Maps

Allele TA Vector (linearized)
pCR2.1-TOPO (linearized)
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pLenti6.3 V5-TOPO (linearized)
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pTargeT (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pENTR5'-TOPO (linearized)
pLUG-Multi (linearized)
pTOP TA V2 (linearized)
pcDNA3.3-TOPO (linearized)
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pMT V5-His-TOPO (linearized)
pTrcHis-TOPO (linearized)
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pOptiVEC-TOPO (linearized)
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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)
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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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