pWJ66

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

Sequence Author: Marraffini Lab / Addgene #46570

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No matches
BsaI (6873) DR BsaI (6776) BsaI (6741) DR pWJ66 9552 bp
BsaI  (6873)
3 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BsaI  (6776)
3 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BsaI  (6741)
3 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
dCas9
2225 .. 6328  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   2225 .. 2251  =  27 bp
   9 amino acids  =  1.1 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
2225 .. 6328  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   2252 .. 2254  =  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
2225 .. 6328  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   2255 .. 4741  =  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
2225 .. 6328  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   4742 .. 4744  =  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
2225 .. 6328  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   4745 .. 6328  =  1584 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
2225 .. 6328  =  4104 bp
1368 amino acids  =  158.3 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
RNAP ω
6335 .. 6607  =  273 bp
90 amino acids  =  10.1 kDa
Product: DNA-directed RNA polymerase subunit omega
promotes RNA polymerase assembly
RNAP ω
6335 .. 6607  =  273 bp
90 amino acids  =  10.1 kDa
Product: DNA-directed RNA polymerase subunit omega
promotes RNA polymerase assembly
CmR
9112 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
9112 .. 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.
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
crRNA leader
6608 .. 6709  =  102 bp
crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system
crRNA leader
6608 .. 6709  =  102 bp
crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system
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
6710 .. 6745  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
DR
6710 .. 6745  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
DR
6776 .. 6811  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
DR
6776 .. 6811  =  36 bp
direct repeat for the Streptococcus pyogenes CRISPR/Cas system
ORF:  763 .. 1059  =  297 bp
ORF:  98 amino acids  =  10.9 kDa
ORF:  2225 .. 6607  =  4383 bp
ORF:  1460 amino acids  =  168.6 kDa
ORF:  7061 .. 7444  =  384 bp
ORF:  127 amino acids  =  14.7 kDa
ORF:  7982 .. 8320  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  8528 .. 8815  =  288 bp
ORF:  95 amino acids  =  10.0 kDa
ORF:  7050 .. 8078  =  1029 bp
ORF:  342 amino acids  =  35.9 kDa
ORF:  9112 .. 219  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  8317 .. 8541  =  225 bp
ORF:  74 amino acids  =  8.2 kDa
ORF:  7125 .. 7367  =  243 bp
ORF:  80 amino acids  =  8.0 kDa
ORF:  7425 .. 7883  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  7241 .. 7591  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
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Download pWJ66.dna file

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