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Plasmid Files

pLenti6.3/TO/V5-DEST

Lentiviral Gateway® destination vector for tetracycline-inducible expression of a gene.

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pLenti6.3 TO V5-DEST Sequence and MappLenti6.3 TO V5-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 KasI (413) PciI (8941) AhdI (8053) DraIII (6913) SfiI (6425) BlpI (5782) PmlI (5318) BclI * (5309) SexAI * (5025) NarI (414) SfoI (415) PluTI (417) EcoNI (942) MfeI (961) AleI (1349) KflI (1706) HpaI (1800) BspDI - ClaI (1923) NdeI (2116) SnaBI (2222) tet operator tet operator SpeI (2498) lac UV5 promoter BspEI (2974) BsmBI (3202) PasI (3210) BstZ17I (3513) SrfI (3849) BmgBI (3883) BfuAI - BspMI (4093) PaeR7I - PspXI - XhoI (4261) XbaI (4267) BstBI (4284) MluI (4328) PfoI (4627) pLenti6.3/TO/V5-DEST 9351 bp
KasI  (413)
1 site
G G C G C C C C G C G G
PciI  (8941)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (8053)
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.
DraIII  (6913)
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  (6425)
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  (5782)
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.
PmlI  (5318)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BclI  (5309)
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.
SexAI  (5025)
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.
NarI  (414)
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  (415)
1 site
G G C G C C C C G C G G
PluTI  (417)
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  (942)
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  (961)
1 site
C A A T T G G T T A A C
AleI  (1349)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (1706)
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  (1800)
1 site
G T T A A C C A A T T G
BspDI  (1923)
1 site
A T C G A T T A G C T A
ClaI  (1923)
1 site
A T C G A T T A G C T A
NdeI  (2116)
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  (2222)
1 site
T A C G T A A T G C A T
SpeI  (2498)
1 site
A C T A G T T G A T C A
BspEI  (2974)
1 site
T C C G G A A G G C C T
BsmBI  (3202)
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.
PasI  (3210)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
BstZ17I  (3513)
1 site
G T A T A C C A T A T G
SrfI  (3849)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (3883)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BfuAI  (4093)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (4093)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PaeR7I  (4261)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4261)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4261)
1 site
C T C G A G G A G C T C
XbaI  (4267)
1 site
T C T A G A A G A T C T
BstBI  (4284)
1 site
T T C G A A A A G C T T
MluI  (4328)
1 site
A C G C G T T G C G C A
PfoI  (4627)
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.
AmpR
7266 .. 8126  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7266 .. 7334  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7266 .. 8126  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7335 .. 8126  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7266 .. 8126  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
2765 .. 3424  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2765 .. 3424  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4354 .. 4942  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4354 .. 4942  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
8297 .. 8885  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
8297 .. 8885  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
6680 .. 7135  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6680 .. 7135  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
5387 .. 5785  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
5387 .. 5785  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
4957 .. 5273  =  317 bp
SV40 enhancer and early promoter
SV40 promoter
4957 .. 5273  =  317 bp
SV40 enhancer and early promoter
ccdB
3766 .. 4071  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
3766 .. 4071  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
RRE
1075 .. 1308  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1075 .. 1308  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
3' LTR (ΔU3)
5872 .. 6105  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
5872 .. 6105  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
RSV promoter
1 .. 229  =  229 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
1 .. 229  =  229 bp
Rous sarcoma virus enhancer/promoter
CMV promoter
2247 .. 2450  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
2247 .. 2450  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
5' LTR (truncated)
230 .. 410  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
230 .. 410  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
457 .. 582  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
457 .. 582  =  126 bp
packaging signal of human immunodeficiency virus
type 1
attR1
2532 .. 2656  =  125 bp
recombination site for the Gateway® LR reaction
attR1
2532 .. 2656  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4112 .. 4236  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4112 .. 4236  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
6177 .. 6298  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6177 .. 6298  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
1804 .. 1921  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
1804 .. 1921  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
7161 .. 7265  =  105 bp
AmpR promoter
7161 .. 7265  =  105 bp
EM7 promoter
5321 .. 5368  =  48 bp
synthetic bacterial promoter
EM7 promoter
5321 .. 5368  =  48 bp
synthetic bacterial promoter
V5 tag
4289 .. 4330  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4289 .. 4330  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
2681 .. 2711  =  31 bp
   Segment 1:  -35  
   2681 .. 2686  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2681 .. 2711  =  31 bp
   Segment 2:  
   2687 .. 2704  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2681 .. 2711  =  31 bp
   Segment 3:  -10  
   2705 .. 2711  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2681 .. 2711  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
tet operator
2452 .. 2470  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2452 .. 2470  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2473 .. 2491  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2473 .. 2491  =  19 bp
bacterial operator O2 for the tetR and tetA genes
SV40 ori
5124 .. 5259  =  136 bp
SV40 origin of replication
SV40 ori
5124 .. 5259  =  136 bp
SV40 origin of replication
TATA box
2436 .. 2442  =  7 bp
TATA box
2436 .. 2442  =  7 bp
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