pLVX-IRES-Neo

Lentiviral vector for bicistronic expression of a gene together with a neomycin (G418) resistance marker.

Sequence Author: Clontech (TaKaRa)

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SgrDI (7949) SspI (7832) PvuI (7398) AhdI (7028) BmtI (5752) NheI (5748) StuI (5737) Bsu36I (4958) PfoI (4511) MluI (4225) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) BstAPI (1453) AleI (1577) KflI - PpuMI (1934) BspDI - ClaI (2180) CMV enhancer NdeI (2374) SnaBI (2480) BsmBI - Esp3I (2729) EcoRI (2803) PaeR7I - XhoI (2809) SpeI (2815) XbaI (2821) NotI (2828) BamHI (2835) PspOMI (2956) ApaI (2960) AvrII (2994) PaqCI (3182) BmgBI (3386) BstXI (3429) MscI (3640) PflFI - Tth111I (3676) SphI (3963) RsrII (4074) pLVX-IRES-Neo 8269 bp
SgrDI  (7949)
1 site
C G T C G A C G G C A G C T G C
SspI  (7832)
1 site
A A T A T T T T A T A A
PvuI  (7398)
1 site
C G A T C G G C T A G C
AhdI  (7028)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BmtI  (5752)
1 site
G C T A G C C G A T C G
NheI  (5748)
1 site
G C T A G C C G A T C G
StuI  (5737)
1 site
A G G C C T T C C G G A
Bsu36I  (4958)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PfoI  (4511)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
MluI  (4225)
1 site
A C G C G T T G C G C A
NruI  (833)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
FseI  (1150)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
EcoNI  (1170)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
MfeI  (1189)
1 site
C A A T T G G T T A A C
BbvCI  (1424)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1424)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BstAPI  (1453)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
AleI  (1577)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (1934)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1934)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BspDI  (2180)
1 site
A T C G A T T A G C T A
ClaI  (2180)
1 site
A T C G A T T A G C T A
NdeI  (2374)
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.
SnaBI  (2480)
1 site
T A C G T A A T G C A T
BsmBI  (2729)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2729)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
EcoRI  (2803)
1 site
G A A T T C C T T A A G
PaeR7I  (2809)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2809)
1 site
C T C G A G G A G C T C
SpeI  (2815)
1 site
A C T A G T T G A T C A
XbaI  (2821)
1 site
T C T A G A A G A T C T
NotI  (2828)
1 site
G C G G C C G C C G C C G G C G
BamHI  (2835)
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  (2956)
1 site
G G G C C C C C C G G G
ApaI  (2960)
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  (2994)
1 site
C C T A G G G G A T C C
PaqCI  (3182)
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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
BmgBI  (3386)
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.
BstXI  (3429)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
MscI  (3640)
1 site
T G G C C A A C C G G T
PflFI  (3676)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3676)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
SphI  (3963)
1 site
G C A T G C C G T A C G
RsrII  (4074)
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
6955 .. 7815  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6955 .. 7746  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6955 .. 7815  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7747 .. 7815  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6955 .. 7815  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
3430 .. 4224  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
3430 .. 4224  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
3' LTR
5033 .. 5666  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5033 .. 5666  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
4238 .. 4826  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4238 .. 4826  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6196 .. 6784  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6196 .. 6784  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
2843 .. 3416  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2843 .. 3416  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
HIV-1 ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
7816 .. 7920  =  105 bp
AmpR promoter
7816 .. 7920  =  105 bp
MCS
2803 .. 2840  =  38 bp
multiple cloning site
MCS
2803 .. 2840  =  38 bp
multiple cloning site
ORF:  3430 .. 4224  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  3326 .. 3601  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  3602 .. 3988  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4328 .. 4855  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  7085 .. 7351  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4341 .. 4637  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  3739 .. 3993  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  6955 .. 7815  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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