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Plasmid Files

pLXIN

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

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pLXIN Sequence and MappLXIN.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SacII (6078) SspI (5736) ScaI (5412) PvuI (5302) AseI (5104) NdeI (3862) BstZ17I (3812) AccI (3811) XbaI (3267) AscI (443) PshAI (577) AfeI (989) BstEII (1153) SexAI * (1281) BsrGI (1348) HpaI (1478) PaeR7I - PspXI - XhoI (1481) BamHI (1486) PspOMI (1645) ApaI (1649) AvrII (1683) HindIII (1756) PmlI (1848) AarI (1871) DraIII (1895) PflMI (1985) BmgBI (2075) SphI (2649) NcoI (2676) NgoMIV (2744) NaeI (2746) RsrII (2760) pLXIN 6098 bp
SacII  (6078)
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  (5736)
1 site
A A T A T T T T A T A A
ScaI  (5412)
1 site
A G T A C T T C A T G A
PvuI  (5302)
1 site
C G A T C G G C T A G C
AseI  (5104)
1 site
A T T A A T T A A T T A
NdeI  (3862)
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  (3812)
1 site
G T A T A C C A T A T G
AccI  (3811)
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  (3267)
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.
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.
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.
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.
PspOMI  (1645)
1 site
G G G C C C C C C G G G
ApaI  (1649)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (1683)
1 site
C C T A G G G G A T C C
HindIII  (1756)
1 site
A A G C T T T T C G A A
PmlI  (1848)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (1871)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
DraIII  (1895)
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.
PflMI  (1985)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BmgBI  (2075)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
SphI  (2649)
1 site
G C A T G C C G T A C G
NcoI  (2676)
1 site
C C A T G G G G T A C C
NgoMIV  (2744)
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  (2746)
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  (2760)
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
4859 .. 5719  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   4859 .. 5650  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4859 .. 5719  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   5651 .. 5719  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4859 .. 5719  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
2107 .. 2910  =  804 bp
267 amino acids  =  29.2 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
2107 .. 2910  =  804 bp
267 amino acids  =  29.2 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
3' LTR
2970 .. 3563  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
3' LTR
2970 .. 3563  =  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
4100 .. 4688  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4100 .. 4688  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
1532 .. 2105  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
1532 .. 2105  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
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
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
5720 .. 5824  =  105 bp
AmpR promoter
5720 .. 5824  =  105 bp
MCS
1476 .. 1491  =  16 bp
multiple cloning site
MCS
1476 .. 1491  =  16 bp
multiple cloning site
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