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pLKO.1 puro

Empty lentiviral vector for expression of shRNA. See also pLKO.1.

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pLKO.1 puro.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Weinberg Lab / Addgene
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EcoRI (6840) MluI (6834) AgeI (6816) BfuAI - BspMI (6815) NdeI (6748) BspDI * - ClaI * (6530) KflI - PpuMI (6441) AleI (6084) BbvCI - Bpu10I (5931) MfeI (5696) EcoNI (5677) NruI * (5344) SphI (4828) PciI (4325) PspFI (4025) BseYI (4021) XcmI (25) SpeI (253) BlpI (333) AscI (342) HincII (480) BamHI (510) PflFI - Tth111I (571) BsiWI (585) BstEII (663) SexAI * (1096) NsiI (1182) Acc65I (1185) KpnI (1189) BbsI (1287) NcoI (1763) SfiI (1809) AvrII (1856) NgoMIV (2189) NaeI (2191) ScaI (2956) pLKO.1 puro 7050 bp
EcoRI  (6840)
1 site
G A A T T C C T T A A G
MluI  (6834)
1 site
A C G C G T T G C G C A
AgeI  (6816)
1 site
A C C G G T T G G C C A
BfuAI  (6815)
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  (6815)
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.
NdeI  (6748)
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.
BspDI  (6530)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (6530)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
KflI  (6441)
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  (6441)
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.
AleI  (6084)
1 site
C A C N N N N G T G G T G N N N N C A C
BbvCI  (5931)
1 site
C C T C A G C G G A G T C G
Bpu10I  (5931)
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.
MfeI  (5696)
1 site
C A A T T G G T T A A C
EcoNI  (5677)
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.
NruI  (5344)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
SphI  (4828)
1 site
G C A T G C C G T A C G
PciI  (4325)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (4025)
1 site
C C C A G C G G G T C G
BseYI  (4021)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
XcmI  (25)
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.
SpeI  (253)
1 site
A C T A G T T G A T C A
BlpI  (333)
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.
AscI  (342)
1 site
G G C G C G C C C C G C G C G G
HincII  (480)
1 site
G T Y R A C C A R Y T G
BamHI  (510)
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.
PflFI  (571)
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  (571)
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.
BsiWI  (585)
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.
BstEII  (663)
1 site
G G