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Plasmid Files

pLPCX

Retroviral vector for transient or stable constitutive expression of a gene.

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pLPCX Sequence and MappLPCX.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SspI (5902) XmnI (5697) PvuI (5468) FspI (5320) AseI (5270) PspFI (4513) BseYI (4509) AflIII - PciI (4205) BspQI - SapI (4089) BspEI * (3927) XbaI (3300) AscI (443) PshAI (577) SpeI (726) BsrGI (1348) BfuAI - BspMI (1478) AgeI (1504) BsiWI (1621) RsrII (1681) DraIII (2136) CMV enhancer BtgZI (2612) SnaBI (2618) BglII (2887) PaeR7I - XhoI (2891) HindIII (2900) BstBI (2905) EcoRI (2907) NotI (2940) BspDI - ClaI (2959) pLPCX 6264 bp
SspI  (5902)
1 site
A A T A T T T T A T A A
XmnI  (5697)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (5468)
1 site
C G A T C G G C T A G C
FspI  (5320)
1 site
T G C G C A A C G C G T
AseI  (5270)
1 site
A T T A A T T A A T T A
PspFI  (4513)
1 site
C C C A G C G G G T C G
BseYI  (4509)
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.
AflIII  (4205)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4205)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4089)
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  (4089)
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.
BspEI  (3927)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
XbaI  (3300)
1 site
T C T A G A A G A T C T
AscI  (443)
1 site
G G C G C G C C C C G C G C G G
PshAI  (577)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SpeI  (726)
1 site
A C T A G T T G A T C A
BsrGI  (1348)
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.
BfuAI  (1478)
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  (1478)
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.
AgeI  (1504)
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.
BsiWI  (1621)
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  (1681)
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.
DraIII  (2136)
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.
BtgZI  (2612)
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.
SnaBI  (2618)
1 site
T A C G T A A T G C A T
BglII  (2887)
1 site
A G A T C T T C T A G A
PaeR7I  (2891)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2891)
1 site
C T C G A G G A G C T C
HindIII  (2900)
1 site
A A G C T T T T C G A A
BstBI  (2905)
1 site
T T C G A A A A G C T T
EcoRI  (2907)
1 site
G A A T T C C T T A A G
NotI  (2940)
1 site
G C G G C C G C C G C C G G C G
BspDI  (2959)
1 site
A T C G A T T A G C T A
ClaI  (2959)
1 site
A T C G A T T A G C T A
AmpR
5025 .. 5885  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   5025 .. 5816  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5025 .. 5885  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5817 .. 5885  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5025 .. 5885  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
1565 .. 2164  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1565 .. 2164  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
3' LTR
3003 .. 3596  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
3' LTR
3003 .. 3596  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
5' LTR
1 .. 589  =  589 bp
long terminal repeat from Moloney murine sarcoma
virus
5' LTR
1 .. 589  =  589 bp
long terminal repeat from Moloney murine sarcoma
virus
ori
4266 .. 4854  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4266 .. 4854  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
gag (truncated)
1052 .. 1468  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
1052 .. 1468  =  417 bp
truncated MMLV gag gene lacking the start codon
CMV enhancer
2339 .. 2642  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2339 .. 2642  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
2643 .. 2846  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
2643 .. 2846  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
MMLV Ψ
652 .. 851  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
652 .. 851  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
AmpR promoter
5886 .. 5990  =  105 bp
AmpR promoter
5886 .. 5990  =  105 bp
MCS
2887 .. 2963  =  77 bp
multiple cloning site
MCS
2887 .. 2963  =  77 bp
multiple cloning site
loxP
3849 .. 3882  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
3849 .. 3882  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
3942 .. 3975  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
3942 .. 3975  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
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