pLVX-Tight-Puro

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

Sequence Author: Clontech (TaKaRa)

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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) CsiI - 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
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