pcDNA-dCas9-p300 Core (D1399Y)

Control plasmid for CRISPR activation, encoding catalytically inactive dCas9 plus an inactivated form of the p300 core transcriptional activation domain.

Sequence Author: Gersbach Lab / Addgene #61358

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HA CMV enhancer ATG SV40 NLS pcDNA-dCas9-p300 Core (D1399Y) 11,475 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
5137 .. 5157  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5137 .. 5157  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
p300 core (D1399Y)
5167 .. 7017  =  1851 bp
617 amino acids  =  71.8 kDa
Product: catalytic histone acetyltransferase core of the human E1A-associated protein p300, inactivated by the D1399Y point mutation (Hilton et al., 2015)
p300 core (D1399Y)
5167 .. 7017  =  1851 bp
617 amino acids  =  71.8 kDa
Product: catalytic histone acetyltransferase core of the human E1A-associated protein p300, inactivated by the D1399Y point mutation (Hilton et al., 2015)
HA
7018 .. 7044  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
7018 .. 7044  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
AmpR
10,479 .. 11,339  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,479 .. 11,270  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,479 .. 11,339  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   11,271 .. 11,339  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,479 .. 11,339  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
8183 .. 8977  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
8183 .. 8977  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
9723 .. 10,308  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9723 .. 10,308  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
7345 .. 7773  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
7345 .. 7773  =  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
7787 .. 8116  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
7787 .. 8116  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
7075 .. 7299  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
7075 .. 7299  =  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
9151 .. 9272  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
9151 .. 9272  =  122 bp
SV40 polyadenylation signal
AmpR promoter
11,340 .. 11,444  =  105 bp
AmpR promoter
11,340 .. 11,444  =  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
7967 .. 8102  =  136 bp
SV40 origin of replication
SV40 ori
7967 .. 8102  =  136 bp
SV40 origin of replication
ORF:  904 .. 7050  =  6147 bp
ORF:  2048 amino acids  =  237.3 kDa
ORF:  10,609 .. 10,875  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  8183 .. 8977  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  7296 .. 7562  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  8355 .. 8741  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  5431 .. 5811  =  381 bp
ORF:  126 amino acids  =  14.4 kDa
ORF:  5977 .. 6270  =  294 bp
ORF:  97 amino acids  =  11.3 kDa
ORF:  6531 .. 6986  =  456 bp
ORF:  151 amino acids  =  16.6 kDa
ORF:  10,479 .. 11,339  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  8492 .. 9028  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
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