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Plasmid Files

pLVX-TRE3G

Lentiviral vector for doxycycline-inducible expression of a gene in cells containing the Tet-On 3G transactivator protein.

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pLVX-TRE3G Sequence and MappLVX-TRE3G.dna
Map and Sequence File   
Sequence Author:  Clontech
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 XcmI (7676) SgrDI (7451) SspI (7334) PvuI (6900) FspI (6752) AhdI (6530) DrdI (5748) PciI (5640) BmtI (5256) NheI (5252) KpnI (4470) Acc65I (4466) Bsu36I (4462) NcoI (4149) PflMI (3982) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI (1424) AleI (1577) KflI (1934) BspDI - ClaI (2180) PaeR7I - PspXI - XhoI (2203) BamHI (2556) PspOMI (2562) ApaI (2566) NotI (2569) TspMI - XmaI (2576) SmaI (2578) MluI (2582) NdeI (2589) EcoRI (2594) SphI (2643) AgeI (2714) SpeI (2849) BlpI (2877) BfuAI - BspMI (3113) PflFI - Tth111I (3166) PasI (3173) BsiWI (3180) RsrII (3240) BstEII (3258) MscI * (3499) SexAI * (3691) pLVX-TRE3G 7819 bp
XcmI  (7676)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SgrDI  (7451)
1 site
C G T C G A C G G C A G C T G C
SspI  (7334)
1 site
A A T A T T T T A T A A
PvuI  (6900)
1 site
C G A T C G G C T A G C
FspI  (6752)
1 site
T G C G C A A C G C G T
AhdI  (6530)
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.
DrdI  (5748)
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.
PciI  (5640)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BmtI  (5256)
1 site
G C T A G C C G A T C G
NheI  (5252)
1 site
G C T A G C C G A T C G
KpnI  (4470)
1 site
G G T A C C C C A T G G
Acc65I  (4466)
1 site
G G T A C C C C A T G G
Bsu36I  (4462)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
NcoI  (4149)
1 site
C C A T G G G G T A C C
PflMI  (3982)
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.
NruI  (833)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
FseI  (1150)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
EcoNI  (1170)
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  (1189)
1 site
C A A T T G G T T A A C
BbvCI  (1424)
1 site
C C T C A G C G G A G T C G
AleI  (1577)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (1934)
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  (2180)
1 site
A T C G A T T A G C T A
ClaI  (2180)
1 site
A T C G A T T A G C T A
PaeR7I  (2203)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2203)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2203)
1 site
C T C G A G G A G C T C
BamHI  (2556)
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.
PspOMI  (2562)
1 site
G G G C C C C C C G G G
ApaI  (2566)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
NotI  (2569)
1 site
G C G G C C G C C G C C G G C G
TspMI  (2576)
1 site
C C C G G G G G G C C C
XmaI  (2576)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (2578)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
MluI  (2582)
1 site
A C G C G T T G C G C A
NdeI  (2589)
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.
EcoRI  (2594)
1 site
G A A T T C C T T A A G
SphI  (2643)
1 site
G C A T G C C G T A C G
AgeI  (2714)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SpeI  (2849)
1 site
A C T A G T T G A T C A
BlpI  (2877)
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.
BfuAI  (3113)
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  (3113)
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.
PflFI  (3166)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3166)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PasI  (3173)
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.
BsiWI  (3180)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
RsrII  (3240)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstEII  (3258)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
MscI  (3499)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
SexAI  (3691)
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.
AmpR
6457 .. 7317  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6457 .. 7248  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6457 .. 7317  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7249 .. 7317  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6457 .. 7317  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
3' LTR
4537 .. 5170  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
4537 .. 5170  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
3124 .. 3723  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3124 .. 3723  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
3741 .. 4329  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
3741 .. 4329  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
5701 .. 6286  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5701 .. 6286  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
2604 .. 3103  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
2604 .. 3103  =  500 bp
mouse phosphoglycerate kinase 1 promoter
TRE3GV promoter
2209 .. 2555  =  347 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On 3G; optimized for
retroviral and lentiviral vectors
TRE3GV promoter
2209 .. 2555  =  347 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On 3G; optimized for
retroviral and lentiviral vectors
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
SV40 poly(A) signal
7470 .. 7604  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7470 .. 7604  =  135 bp
SV40 polyadenylation signal
HIV-1 ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
AmpR promoter
7318 .. 7422  =  105 bp
AmpR promoter
7318 .. 7422  =  105 bp
MCS
2556 .. 2599  =  44 bp
multiple cloning site
MCS
2556 .. 2599  =  44 bp
multiple cloning site
tet operator
2214 .. 2232  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2214 .. 2232  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2250 .. 2268  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2250 .. 2268  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2286 .. 2304  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2286 .. 2304  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2322 .. 2340  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2322 .. 2340  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2358 .. 2376  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2358 .. 2376  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2394 .. 2412  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2394 .. 2412  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2430 .. 2448  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2430 .. 2448  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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