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pLVX-PTuner Green

Lentiviral vector for fusing a destabilization domain to the N-terminus of a partner protein, with co-expression of ZsGreen1.

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pLVX-PTuner Green.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SgrDI (8169) SspI (8052) PvuI (7618) FspI (7470) DrdI (6463) BmtI (5972) NheI (5968) StuI (5957) Bsu36I (5178) PstI (5063) PfoI (4731) MluI (4445) BsrGI (4299) BssHII (711) NruI * (833) FseI (1150) MfeI (1189) BbvCI - Bpu10I (1424) BstAPI (1453) AleI (1577) KflI - PpuMI (1934) BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) SwaI (2797) TspMI - XmaI (2923) SmaI (2925) AgeI (3129) EcoRI (3135) PaeR7I - XhoI (3141) SpeI (3147) XbaI (3153) NotI (3160) BamHI (3167) PspOMI (3288) ApaI (3292) AvrII (3326) BclI * (3831) SgrAI (3836) PflFI - Tth111I (3983) pLVX-PTuner Green 8536 bp
SgrDI  (8169)
1 site
C G T C G A C G G C A G C T G C
SspI  (8052)
1 site
A A T A T T T T A T A A
PvuI  (7618)
1 site
C G A T C G G C T A G C
FspI  (7470)
1 site
T G C G C A A C G C G T
DrdI  (6463)
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.
BmtI  (5972)
1 site
G C T A G C C G A T C G
NheI  (5968)
1 site
G C T A G C C G A T C G
StuI  (5957)
1 site
A G G C C T T C C G G A
Bsu36I  (5178)
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.
PstI  (5063)
1 site
C T G C A G G A C G T C
PfoI  (4731)
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.
MluI  (4445)
1 site
A C G C G T T G C G C A
BsrGI  (4299)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BssHII  (711)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
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.
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
Bpu10I  (1424)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
BstAPI  (1453)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
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.
PpuMI  (1934)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI 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
NdeI  (2374)
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  (2480)
1 site
T A C G T A A T G C A T
SwaI  (2797)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
TspMI  (2923)
1 site
C C C G G G G G G C C C
XmaI  (2923)
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  (2925)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
AgeI  (3129)
1 site
A C C G G T T G G C C A
EcoRI  (3135)
1 site
G A A T T C C T T A A G
PaeR7I  (3141)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3141)
1 site
C T C G A G G A G C T C
SpeI  (3147)
1 site
A C T A G T T G A T C A
XbaI  (3153)
1 site
T C T A G A A G A T C T
NotI  (3160)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3167)
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  (3288)
1 site
G G G C C C C C C G G G
ApaI  (3292)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (3326)
1 site
C C T A G G G G A T C C
BclI  (3831)
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.
SgrAI  (3836)
1 site
C R C C G G Y G