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Plasmid Files

pLP2

Lentiviral packaging plasmid containing the HIV-1 rev gene.

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pLP2 Sequence and MappLP2.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 Bsu36I (4141) MluI (4069) BspQI - SapI (3807) PciI (3690) PspFI (3390) BseYI (3386) AhdI (2802) BmrI (2762) BsaI (2736) NmeAIII (2655) ScaI (2321) XmnI (2202) SphI (93) AleI (251) PaeR7I - PspXI - XhoI (269) Eco53kI (274) SacI (276) BseRI (415) KflI - PpuMI (488) BamHI (563) BbvCI (780) BglII (864) AflII (893) KasI (1099) NarI (1100) SfoI (1101) PluTI (1103) NgoMIV (1298) NaeI (1300) BtgZI (1398) BsaAI (1403) DraIII (1406) PsiI (1531) PfoI (1763) ZraI (1881) AatII (1883) pLP2 4180 bp
Bsu36I  (4141)
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.
MluI  (4069)
1 site
A C G C G T T G C G C A
BspQI  (3807)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3807)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PciI  (3690)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (3390)
1 site
C C C A G C G G G T C G
BseYI  (3386)
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.
AhdI  (2802)
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.
BmrI  (2762)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
BsaI  (2736)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
NmeAIII  (2655)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
ScaI  (2321)
1 site
A G T A C T T C A T G A
XmnI  (2202)
1 site
G A A N N N N T T C C T T N N N N A A G
SphI  (93)
1 site
G C A T G C C G T A C G
AleI  (251)
1 site
C A C N N N N G T G G T G N N N N C A C
PaeR7I  (269)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (269)
1 site
V C T C G A G B B G A G C T C V
XhoI  (269)
1 site
C T C G A G G A G C T C
Eco53kI  (274)
1 site
G A G C T C C T C G A G
SacI  (276)
1 site
G A G C T C C T C G A G
BseRI  (415)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
KflI  (488)
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  (488)
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.
BamHI  (563)
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.
BbvCI  (780)
1 site
C C T C A G C G G A G T C G
BglII  (864)
1 site
A G A T C T T C T A G A
AflII  (893)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
KasI  (1099)
1 site
G G C G C C C C G C G G
NarI  (1100)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (1101)
1 site
G G C G C C C C G C G G
PluTI  (1103)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
NgoMIV  (1298)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (1300)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
BtgZI  (1398)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (1403)
1 site
Y A C G T R R T G C A Y
DraIII  (1406)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
PsiI  (1531)
1 site
T T A T A A A A T A T T
PfoI  (1763)
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.
ZraI  (1881)
1 site
G A C G T C C T G C A G
AatII  (1883)
1 site
G A C G T C C T G C A G
AmpR
2015 .. 2875  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2015 .. 2083  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2015 .. 2875  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2084 .. 2875  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2015 .. 2875  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3046 .. 3634  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3046 .. 3634  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1173 .. 1628  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1173 .. 1628  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rev
391 .. 741  =  351 bp
116 amino acids  =  13.1 kDa
Product: HIV-1 Rev protein
rev
391 .. 741  =  351 bp
116 amino acids  =  13.1 kDa
Product: HIV-1 Rev protein
RSV promoter
1 .. 262  =  262 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
1 .. 262  =  262 bp
Rous sarcoma virus enhancer/promoter
HIV-1 poly(A)
850 .. 971  =  122 bp
polyadenylation signal from the LTR of HIV-1
HIV-1 poly(A)
850 .. 971  =  122 bp
polyadenylation signal from the LTR of HIV-1
AmpR promoter
1910 .. 2014  =  105 bp
AmpR promoter
1910 .. 2014  =  105 bp
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