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Plasmid Files

pLX-sgRNA

Lentiviral vector for expressing single guide RNA (sgRNA) sequences generated by overlap extension PCR.

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pLX-sgRNA Sequence and MappLX-sgRNA.dna
Map and Sequence File   
Sequence Author:  Lander & Sabatini Labs / Addgene #50662
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 R2 (3275 .. 3307) F1 (2841 .. 2870) XhoI (2841) gRNA scaffold NheI (3302) pLX-sgRNA 7477 bp
XhoI  (2841)
1 site
C T C G A G G A G C T C
NheI  (3302)
1 site
G C T A G C C G A T C G
R2
36-mer  /  39% GC
1 binding site
3275 .. 3307  =  33 annealed bases
Tm  =  62°C
overlap extension PCR primer with an NheI site
F1
33-mer  /  36% GC
1 binding site
2841 .. 2870  =  30 annealed bases
Tm  =  61°C
overlap extension PCR primer with an XhoI site
AmpR
5394 .. 6254  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5394 .. 5462  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5394 .. 6254  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5463 .. 6254  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5394 .. 6254  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
WPRE
3334 .. 3922  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
3334 .. 3922  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6425 .. 7013  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6425 .. 7013  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hPGK promoter
1714 .. 2214  =  501 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
1714 .. 2214  =  501 bp
human phosphoglycerate kinase 1 promoter
f1 ori
4808 .. 5263  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4808 .. 5263  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
2226 .. 2621  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
2226 .. 2621  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
U6 promoter
2916 .. 3156  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2916 .. 3156  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1077 .. 1310  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1077 .. 1310  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
3' LTR (ΔU3)
3994 .. 4227  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
3994 .. 4227  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
RSV promoter
5 .. 231  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
5 .. 231  =  227 bp
Rous sarcoma virus enhancer/promoter
5' LTR (truncated)
232 .. 412  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
232 .. 412  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
4466 .. 4601  =  136 bp
SV40 origin of replication
SV40 ori
4466 .. 4601  =  136 bp
SV40 origin of replication
HIV-1 Ψ
459 .. 584  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
459 .. 584  =  126 bp
packaging signal of human immunodeficiency virus
type 1
SV40 poly(A) signal
4305 .. 4426  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4305 .. 4426  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2681 .. 2798  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2681 .. 2798  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
5289 .. 5393  =  105 bp
AmpR promoter
5289 .. 5393  =  105 bp
gRNA scaffold
3185 .. 3260  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
3185 .. 3260  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
polIII terminator
3261 .. 3266  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3261 .. 3266  =  6 bp
RNA polymerase III transcription terminator
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