pcDNA-dCas9

Plasmid for expressing catalytically inactive dCas9 in mammalian cells using the CMV promoter.

Sequence Author: Gersbach Lab / Addgene #47106

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CMV enhancer ATG SV40 NLS pcDNA-dCas9 9575 bp
ATG
904 .. 906  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
904 .. 906  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
907 .. 972  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
907 .. 972  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
979 .. 999  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
979 .. 999  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
dCas9
1009 .. 5112  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   1009 .. 1035  =  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
1009 .. 5112  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   1036 .. 1038  =  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
1009 .. 5112  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   1039 .. 3525  =  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
1009 .. 5112  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   3526 .. 3528  =  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
1009 .. 5112  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   3529 .. 5112  =  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
1009 .. 5112  =  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
SV40 NLS
5113 .. 5133  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5113 .. 5133  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
AmpR
8579 .. 9439  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8579 .. 9370  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8579 .. 9439  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   9371 .. 9439  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8579 .. 9439  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
6283 .. 7077  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
6283 .. 7077  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
7823 .. 8408  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7823 .. 8408  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5445 .. 5873  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5445 .. 5873  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
5887 .. 6216  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
5887 .. 6216  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
5175 .. 5399  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5175 .. 5399  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
7251 .. 7372  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7251 .. 7372  =  122 bp
SV40 polyadenylation signal
AmpR promoter
9440 .. 9544  =  105 bp
AmpR promoter
9440 .. 9544  =  105 bp
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
6067 .. 6202  =  136 bp
SV40 origin of replication
SV40 ori
6067 .. 6202  =  136 bp
SV40 origin of replication
ORF:  904 .. 5139  =  4236 bp
ORF:  1411 amino acids  =  163.4 kDa
ORF:  6283 .. 7077  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5396 .. 5662  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  6455 .. 6841  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  8709 .. 8975  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  8579 .. 9439  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  6592 .. 7128  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
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