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Plasmid Files

pLNCX2

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

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pLNCX2 Sequence and MappLNCX2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SacII (6112) SspI (5800) XmnI (5595) ScaI (5476) PvuI (5366) AseI (5168) AflIII - PciI (4103) BstZ17I (3876) XbaI (3331) 3' pLNCX Seq/PCR primer (3031 .. 3056) AscI (443) PshAI (577) SpeI (726) BstEII (1153) SexAI * (1281) BsrGI (1348) BclI * (1481) BsaBI * (1499) SphI (2045) NgoMIV (2140) NaeI (2142) RsrII (2156) BstBI (2322) CMV enhancer SnaBI (2656) 5' pLNCX Seq/PCR primer (2881 .. 2905) BglII (2925) PaeR7I - XhoI (2929) HindIII (2938) NotI (2956) SalI (2965) StuI (2973) BspDI - ClaI (2990) pLNCX2 6132 bp
SacII  (6112)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
SspI  (5800)
1 site
A A T A T T T T A T A A
XmnI  (5595)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5476)
1 site
A G T A C T T C A T G A
PvuI  (5366)
1 site
C G A T C G G C T A G C
AseI  (5168)
1 site
A T T A A T T A A T T A
AflIII  (4103)
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  (4103)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3876)
1 site
G T A T A C C A T A T G
XbaI  (3331)
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
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.
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.
BclI  (1481)
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.
BsaBI  (1499)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
SphI  (2045)
1 site
G C A T G C C G T A C G
NgoMIV  (2140)
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  (2142)
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.
RsrII  (2156)
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.
BstBI  (2322)
1 site
T T C G A A A A G C T T
SnaBI  (2656)
1 site
T A C G T A A T G C A T
BglII  (2925)
1 site
A G A T C T T C T A G A
PaeR7I  (2929)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2929)
1 site
C T C G A G G A G C T C
HindIII  (2938)
1 site
A A G C T T T T C G A A
NotI  (2956)
1 site
G C G G C C G C C G C C G G C G
SalI  (2965)
1 site
G T C G A C C A G C T G
StuI  (2973)
1 site
A G G C C T T C C G G A
BspDI  (2990)
1 site
A T C G A T T A G C T A
ClaI  (2990)
1 site
A T C G A T T A G C T A
3' pLNCX Seq/PCR primer
26-mer  /  54% GC
1 binding site
3031 .. 3056  =  26 annealed bases
Tm  =  64°C
5' pLNCX Seq/PCR primer
25-mer  /  48% GC
1 binding site
2881 .. 2905  =  25 annealed bases
Tm  =  61°C
AmpR
4923 .. 5783  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4923 .. 5714  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4923 .. 5783  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5715 .. 5783  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4923 .. 5783  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
1512 .. 2306  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1512 .. 2306  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
3' LTR
3034 .. 3627  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
3' LTR
3034 .. 3627  =  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
4164 .. 4752  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4164 .. 4752  =  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
2377 .. 2680  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2377 .. 2680  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
2681 .. 2884  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
2681 .. 2884  =  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
5784 .. 5888  =  105 bp
AmpR promoter
5784 .. 5888  =  105 bp
MCS
2925 .. 2994  =  70 bp
multiple cloning site
MCS
2925 .. 2994  =  70 bp
multiple cloning site
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