Resources
Plasmid Files

pLX-sgRNA

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

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pLX-sgRNA.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Lander & Sabatini Labs / Addgene #50662
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F1 (2841 .. 2870) XhoI (2841) NheI (3302) R2 (3275 .. 3307) 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
F1
33-mer  /  36% GC
1 binding site
2841 .. 2870  =  30 annealed bases
Tm  =  61°C
overlap extension PCR primer with an XhoI site
R2
36-mer  /  39% GC
1 binding site
3275 .. 3307  =  33 annealed bases
Tm  =  62°C
overlap extension PCR primer with an NheI 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
ORF:  955 .. 1938  =  984 bp
ORF:  327 amino acids  =  36.7 kDa
ORF:  1996 .. 2445  =  450 bp
ORF:  149 amino acids  =  15.8 kDa
ORF:  3424 .. 3933  =  510 bp
ORF:  169 amino acids  =  18.5 kDa
ORF:  4327 .. 4563  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  7411 .. 179  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  3437 .. 3967  =  531 bp
ORF:  176 amino acids  =  18.7 kDa
ORF:  2226 .. 2636  =  411 bp
ORF:  136 amino acids  =  14.2 kDa
ORF:  2661 .. 3029  =  369 bp
ORF:  122 amino acids  =  14.1 kDa
ORF:  5394 .. 6254  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7338 .. 160  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  5858 .. 6124  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  870 .. 1250  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  1683 .. 2528  =  846 bp
ORF:  281 amino acids  =  30.3 kDa
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