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Plasmid Files

pLXSN

Retroviral vector for co-expression of a gene together with a neomycin (G418) resistance marker.

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pLXSN Sequence and MappLXSN.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SacII (5854) SspI (5512) XmnI (5307) ScaI (5188) PvuI (5078) AseI (4880) AflIII - PciI (3815) NdeI (3638) BstZ17I (3588) AccI (3587) XbaI (3043) AscI (443) PshAI (577) SpeI (726) AfeI (989) BstEII (1153) BsrGI (1348) EcoRI (1470) HpaI (1478) PaeR7I - PspXI - XhoI (1481) BamHI (1486) SfiI (1770) StuI (1816) AvrII (1817) HindIII (1833) BclI * (1861) BsaBI * (1879) NgoMIV (2520) NaeI (2522) RsrII (2536) pLXSN 5874 bp
SacII  (5854)
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  (5512)
1 site
A A T A T T T T A T A A
XmnI  (5307)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5188)
1 site
A G T A C T T C A T G A
PvuI  (5078)
1 site
C G A T C G G C T A G C
AseI  (4880)
1 site
A T T A A T T A A T T A
AflIII  (3815)
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  (3815)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NdeI  (3638)
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.
BstZ17I  (3588)
1 site
G T A T A C C A T A T G
AccI  (3587)
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.
XbaI  (3043)
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
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.
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.
EcoRI  (1470)
1 site
G A A T T C C T T A A G
HpaI  (1478)
1 site
G T T A A C C A A T T G
PaeR7I  (1481)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1481)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1481)
1 site
C T C G A G G A G C T C
BamHI  (1486)
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.
SfiI  (1770)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
StuI  (1816)
1 site
A G G C C T T C C G G A
AvrII  (1817)
1 site
C C T A G G G G A T C C
HindIII  (1833)
1 site
A A G C T T T T C G A A
BclI  (1861)
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  (1879)
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.
NgoMIV  (2520)
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  (2522)
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  (2536)
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.
AmpR
4635 .. 5495  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4635 .. 5426  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4635 .. 5495  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5427 .. 5495  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4635 .. 5495  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
1892 .. 2686  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1892 .. 2686  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
3' LTR
2746 .. 3339  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
3' LTR
2746 .. 3339  =  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
3876 .. 4464  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3876 .. 4464  =  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
SV40 promoter
1503 .. 1832  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1503 .. 1832  =  330 bp
SV40 enhancer and 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
5496 .. 5600  =  105 bp
AmpR promoter
5496 .. 5600  =  105 bp
MCS
1470 .. 1491  =  22 bp
multiple cloning site
MCS
1470 .. 1491  =  22 bp
multiple cloning site
SV40 ori
1683 .. 1818  =  136 bp
SV40 origin of replication
SV40 ori
1683 .. 1818  =  136 bp
SV40 origin of replication
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