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Plasmid Files

pLVX-IRES-Hyg

Lentiviral vector for bicistronic expression of a gene together with a hygromycin resistance marker.

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pLVX-IRES-Hyg Sequence and MappLVX-IRES-Hyg.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SgrDI (8199) SspI (8082) FspI (7500) AhdI (7278) BmtI (6002) NheI (5998) StuI (5987) Bsu36I (5208) PfoI (4761) MluI (4475) KasI (637) NarI (638) SfoI (639) PluTI (641) BssHII (711) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) AleI (1577) KflI - PpuMI (1934) BspDI - ClaI (2180) CMV enhancer SnaBI (2480) PaeR7I - XhoI (2809) SpeI (2815) XbaI (2821) NotI (2828) BamHI (2835) PspOMI (2956) ApaI (2960) AvrII (2994) AarI (3182) BmgBI (3386) PshAI (3476) AsiSI (3813) RsrII (3857) pLVX-IRES-Hyg 8519 bp
SgrDI  (8199)
1 site
C G T C G A C G G C A G C T G C
SspI  (8082)
1 site
A A T A T T T T A T A A
FspI  (7500)
1 site
T G C G C A A C G C G T
AhdI  (7278)
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  (6002)
1 site
G C T A G C C G A T C G
NheI  (5998)
1 site
G C T A G C C G A T C G
StuI  (5987)
1 site
A G G C C T T C C G G A
Bsu36I  (5208)
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  (4761)
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  (4475)
1 site
A C G C G T T G C G C A
KasI  (637)
1 site
G G C G C C C C G C G G
NarI  (638)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (639)
1 site
G G C G C C C C G C G G
PluTI  (641)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
BssHII  (711)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
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
SnaBI  (2480)
1 site
T A C G T A A T G C A T
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
AarI  (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 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.
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.
PshAI  (3476)
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.
AsiSI  (3813)
1 site
G C G A T C G C C G C T A G C G
RsrII  (3857)
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.
HygR
3455 .. 4474  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
3455 .. 4474  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
7205 .. 8065  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7205 .. 7996  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7205 .. 8065  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7997 .. 8065  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7205 .. 8065  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
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
5283 .. 5916  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5283 .. 5916  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
4488 .. 5076  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4488 .. 5076  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6446 .. 7034  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6446 .. 7034  =  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
8066 .. 8170  =  105 bp
AmpR promoter
8066 .. 8170  =  105 bp
MCS
2803 .. 2840  =  38 bp
multiple cloning site
MCS
2803 .. 2840  =  38 bp
multiple cloning site
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