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Plasmid Files

pLVX-Tight-Puro

Lentiviral vector for expressing a gene with the Tet-On® Advanced or Tet-Off® Advanced system.

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pLVX-Tight-Puro Sequence and MappLVX-Tight-Puro.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SgrDI (7423) SspI (7306) PvuI (6872) FspI (6724) AhdI (6502) PciI (5609) BmtI (5226) NheI (5222) KpnI (4440) Acc65I (4436) Bsu36I (4432) NcoI (4120) PflMI (3953) FseI (1150) MfeI (1189) BbvCI (1424) AleI (1577) KflI (1934) BspDI - ClaI (2180) PaeR7I - PspXI - XhoI (2203) BamHI (2528) NotI (2535) XbaI * (2548) MluI (2560) EcoRI (2566) SphI (2615) AgeI (2686) SpeI (2821) BlpI (2849) BfuAI - BspMI (3085) PflFI - Tth111I (3138) PasI (3145) BsiWI (3152) RsrII (3212) BstEII (3230) MscI * (3471) SexAI * (3663) pLVX-Tight-Puro 7790 bp
SgrDI  (7423)
1 site
C G T C G A C G G C A G C T G C
SspI  (7306)
1 site
A A T A T T T T A T A A
PvuI  (6872)
1 site
C G A T C G G C T A G C
FspI  (6724)
1 site
T G C G C A A C G C G T
AhdI  (6502)
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.
PciI  (5609)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BmtI  (5226)
1 site
G C T A G C C G A T C G
NheI  (5222)
1 site
G C T A G C C G A T C G
KpnI  (4440)
1 site
G G T A C C C C A T G G
Acc65I  (4436)
1 site
G G T A C C C C A T G G
Bsu36I  (4432)
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  (4120)
1 site
C C A T G G G G T A C C
PflMI  (3953)
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.
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.
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  (2528)
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.
NotI  (2535)
1 site
G C G G C C G C C G C C G G C G
XbaI  (2548)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MluI  (2560)
1 site
A C G C G T T G C G C A
EcoRI  (2566)
1 site
G A A T T C C T T A A G
SphI  (2615)
1 site
G C A T G C C G T A C G
AgeI  (2686)
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  (2821)
1 site
A C T A G T T G A T C A
BlpI  (2849)
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  (3085)
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  (3085)
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  (3138)
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  (3138)
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  (3145)
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  (3152)
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  (3212)
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  (3230)
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  (3471)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
SexAI  (3663)
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
6429 .. 7289  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6429 .. 7220  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6429 .. 7289  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7221 .. 7289  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6429 .. 7289  =  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
4507 .. 5140  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
4507 .. 5140  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
3096 .. 3695  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3096 .. 3695  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
3712 .. 4300  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
3712 .. 4300  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
5670 .. 6258  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5670 .. 6258  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
2576 .. 3075  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
2576 .. 3075  =  500 bp
mouse phosphoglycerate kinase 1 promoter
tight TRE promoter
2206 .. 2520  =  315 bp
Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter
tight TRE promoter
2206 .. 2520  =  315 bp
Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter
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
7442 .. 7576  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7442 .. 7576  =  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
7290 .. 7394  =  105 bp
AmpR promoter
7290 .. 7394  =  105 bp
MCS
2528 .. 2571  =  44 bp
multiple cloning site
MCS
2528 .. 2571  =  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
2285 .. 2303  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2285 .. 2303  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2321 .. 2339  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2321 .. 2339  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2357 .. 2375  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2357 .. 2375  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2392 .. 2410  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2392 .. 2410  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2428 .. 2446  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2428 .. 2446  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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