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Plasmid Files

pLVX-hyg-sgRNA1 Vector (Linear)

Linearized lentiviral vector with a hygromycin resistance marker, for cloning and expression of a single guide RNA (sgRNA).

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pLVX-hyg-sgRNA1 Vector (Linear).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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End (8219) BspDI - ClaI (7955) KflI (7709) AleI (7352) BbvCI (7199) MfeI (6964) EcoNI (6945) FseI (6925) NruI * (6608) BssHII (6486) XcmI (5632) SgrDI (5407) SspI (5290) FspI (4708) Start (0) SphI (166) AgeI (237) SpeI (372) BlpI (400) PshAI (671) AsiSI (1008) RsrII (1052) PflFI - Tth111I (1148) PspOMI (1676) ApaI (1680) PflMI (1938) PfoI (1970) Bsu36I (2418) Acc65I (2422) KpnI (2426) NheI (3208) BmtI (3212) PciI (3596) pLVX-hyg-sgRNA1 Vector (Linear) 8219 bp
End  (8219)
0 sites
BspDI  (7955)
1 site
A T C G A T T A G C T A
ClaI  (7955)
1 site
A T C G A T T A G C T A
KflI  (7709)
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.
AleI  (7352)
1 site
C A C N N N N G T G G T G N N N N C A C
BbvCI  (7199)
1 site
C C T C A G C G G A G T C G
MfeI  (6964)
1 site
C A A T T G G T T A A C
EcoNI  (6945)
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.
FseI  (6925)
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.
NruI  (6608)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
BssHII  (6486)
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.
XcmI  (5632)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SgrDI  (5407)
1 site
C G T C G A C G G C A G C T G C
SspI  (5290)
1 site
A A T A T T T T A T A A
FspI  (4708)
1 site
T G C G C A A C G C G T
Start  (0)
0 sites
SphI  (166)
1 site
G C A T G C C G T A C G
AgeI  (237)
1 site
A C C G G T T G G C C A
SpeI  (372)
1 site
A C T A G T T G A T C A
BlpI  (400)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
PshAI  (671)
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  (1008)
1 site
G C G A T C G C C G C T A G C G
RsrII  (1052)
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.
PflFI  (1148)
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  (1148)
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.
PspOMI  (1676)
1 site
G G G C C C C C C G G G
ApaI  (1680)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PflMI  (1938)
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.
PfoI  (1970)
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.
Bsu36I  (2418)
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.
Acc65I  (2422)
1 site
G G T A C C C C A T G G
KpnI  (2426)
1 site
G G T A C C C C A T G G
NheI  (3208)
1 site
G C T A G C C G A T C G
BmtI  (3212)
1 site
G C T A G C C G A T C G
PciI  (3596)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
HygR
644 .. 1669  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
644 .. 1669  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
AmpR
4413 .. 5273  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4413 .. 5204  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4413 .. 5273  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5205 .. 5273  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4413 .. 5273  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
3' LTR
2493 .. 3126  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
2493 .. 3126  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5776 .. 6409  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5776 .. 6409  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
1697 .. 2285  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
1697 .. 2285  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
3657 .. 4242  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3657 .. 4242  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
127 .. 626  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
127 .. 626  =  500 bp
mouse phosphoglycerate kinase 1 promoter
U6 promoter
7975 .. 8215  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
7975 .. 8215  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
7078 .. 7311  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
7078 .. 7311  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
SV40 poly(A) signal
5426 .. 5560  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5426 .. 5560  =  135 bp
SV40 polyadenylation signal
HIV-1 Ψ
6456 .. 6581  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
6456 .. 6581  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
7803 .. 7918  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
7803 .. 7918  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
5274 .. 5378  =  105 bp
AmpR promoter
5274 .. 5378  =  105 bp
gRNA scaffold
1 .. 86  =  86 bp
improved guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
1 .. 86  =  86 bp
improved guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
ORF:  4543 .. 4809  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  407 .. 1669  =  1263 bp
ORF:  420 amino acids  =  46.5 kDa
ORF:  1787 .. 2314  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  6956 .. 7807  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  1800 .. 2096  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  6790 .. 7251  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  4413 .. 5273  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  572 .. 1402  =  831 bp
ORF:  276 amino acids  =  31.0 kDa
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