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Plasmid Files

pLHCX

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

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pLHCX Sequence and MappLHCX.dna
Map and Sequence File   
Sequence Author:  Clontech
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 XmnI (6276) FspI (5899) AflIII - PciI (4784) BspQI - SapI (4668) BstZ17I (4557) AccI (4556) 3' pLNCX Seq/PCR primer (3712 .. 3737) BspDI - ClaI (3671) HpaI (3666) HindIII (3658) 5' pLNCX Seq/PCR primer (3561 .. 3585) AscI (443) AfeI (989) BstEII (1153) SexAI * (1281) BfuAI - BspMI (1831) AsiSI (1882) RsrII (1926) BlpI (2669) AleI (2769) BglII (2829) BsgI (2846) CMV enhancer SnaBI (3336) pLHCX 6813 bp
XmnI  (6276)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (5899)
1 site
T G C G C A A C G C G T
AflIII  (4784)
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  (4784)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4668)
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  (4668)
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.
BstZ17I  (4557)
1 site
G T A T A C C A T A T G
AccI  (4556)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BspDI  (3671)
1 site
A T C G A T T A G C T A
ClaI  (3671)
1 site
A T C G A T T A G C T A
HpaI  (3666)
1 site
G T T A A C C A A T T G
HindIII  (3658)
1 site
A A G C T T T T C G A A
AscI  (443)
1 site
G G C G C G C C C C G C G C G G
AfeI  (989)
1 site
A G C G C T T C G C G A
BstEII  (1153)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
SexAI  (1281)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BfuAI  (1831)
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  (1831)
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.
AsiSI  (1882)
1 site
G C G A T C G C C G C T A G C G
RsrII  (1926)
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.
BlpI  (2669)
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.
AleI  (2769)
1 site
C A C N N N N G T G G T G N N N N C A C
BglII  (2829)
1 site
A G A T C T T C T A G A
BsgI  (2846)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
SnaBI  (3336)
1 site
T A C G T A A T G C A T
3' pLNCX Seq/PCR primer
26-mer  /  54% GC
1 binding site
3712 .. 3737  =  26 annealed bases
Tm  =  64°C
5' pLNCX Seq/PCR primer
25-mer  /  48% GC
1 binding site
3561 .. 3585  =  25 annealed bases
Tm  =  60°C
HygR
1524 .. 2543  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
1524 .. 2543  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
5604 .. 6464  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   5604 .. 6395  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5604 .. 6464  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   6396 .. 6464  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5604 .. 6464  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
3' LTR
3715 .. 4308  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
3' LTR
3715 .. 4308  =  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
4845 .. 5433  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4845 .. 5433  =  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
2981 .. 3360  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2981 .. 3360  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
3361 .. 3564  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
3361 .. 3564  =  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
6465 .. 6569  =  105 bp
AmpR promoter
6465 .. 6569  =  105 bp
MCS
3658 .. 3675  =  18 bp
multiple cloning site
MCS
3658 .. 3675  =  18 bp
multiple cloning site
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