pWJ68

Bacterial plasmid for expressing catalytically inactive dCas9, with an N-terminal fusion to the RNA polymerase ω subunit for transcriptional activation.

Sequence Author: Marraffini Lab / Addgene #46571

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DR DR pWJ68 9622 bp
RNAP ω
2245 .. 2517  =  273 bp
91 amino acids  =  10.2 kDa
Product: DNA-directed RNA polymerase subunit omega
promotes RNA polymerase assembly
RNAP ω
2245 .. 2517  =  273 bp
91 amino acids  =  10.2 kDa
Product: DNA-directed RNA polymerase subunit omega
promotes RNA polymerase assembly
dCas9
2524 .. 6627  =  4104 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   2524 .. 2547  =  24 bp
   8 amino acids  =  923.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2524 .. 6627  =  4104 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   2548 .. 2550  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2524 .. 6627  =  4104 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   2551 .. 5037  =  2487 bp
   829 amino acids  =  96.3 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2524 .. 6627  =  4104 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   5038 .. 5040  =  3 bp
   1 amino acid  =  89.1 Da
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2524 .. 6627  =  4104 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   5041 .. 6627  =  1587 bp
   528 amino acids  =  60.9 kDa
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
dCas9
2524 .. 6627  =  4104 bp
1367 amino acids  =  158.2 kDa
5 segments
Product: catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain
CmR
9182 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
9182 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
p15A ori
848 .. 1393  =  546 bp
Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin.
p15A ori
848 .. 1393  =  546 bp
Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin.
crRNA leader
6648 .. 6779  =  132 bp
crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system
crRNA leader
6648 .. 6779  =  132 bp
crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system
cat promoter
220 .. 322  =  103 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
cat promoter
220 .. 322  =  103 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
tracrRNA
1851 .. 1929  =  79 bp
trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system
tracrRNA
1851 .. 1929  =  79 bp
trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system
DR
6780 .. 6815  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
DR
6780 .. 6815  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
DR
6846 .. 6881  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
DR
6846 .. 6881  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
ORF:  763 .. 1059  =  297 bp
ORF:  98 amino acids  =  10.9 kDa
ORF:  2245 .. 6627  =  4383 bp
ORF:  1460 amino acids  =  168.6 kDa
ORF:  7120 .. 8148  =  1029 bp
ORF:  342 amino acids  =  35.9 kDa
ORF:  7131 .. 7514  =  384 bp
ORF:  127 amino acids  =  14.7 kDa
ORF:  8052 .. 8390  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  8598 .. 8885  =  288 bp
ORF:  95 amino acids  =  10.0 kDa
ORF:  8387 .. 8611  =  225 bp
ORF:  74 amino acids  =  8.2 kDa
ORF:  9182 .. 219  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  7195 .. 7437  =  243 bp
ORF:  80 amino acids  =  8.0 kDa
ORF:  7495 .. 7953  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  7311 .. 7661  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
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