pAC94-pmax-dCas9VP160-2A-puro

CRISPR activation vector for expressing a puromycin resistance marker plus catalytically inactive dCas9 fused to the VP160 transcriptional activation domain.

Sequence Author: Jaenisch Lab / Addgene #48226

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CMV enhancer attB1 ATG HA SV40 NLS pAC94-pmax-dCas9VP160-2A-puro 8282 bp
ATG
1005 .. 1007  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1005 .. 1007  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
HA
1008 .. 1034  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
1008 .. 1034  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
SV40 NLS
1038 .. 1058  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1038 .. 1058  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
dCas9
1068 .. 5168  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   1068 .. 1091  =  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
1068 .. 5168  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   1092 .. 1094  =  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
1068 .. 5168  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   1095 .. 3581  =  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
1068 .. 5168  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   3582 .. 3584  =  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
1068 .. 5168  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   3585 .. 5168  =  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
1068 .. 5168  =  4101 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
SV40 NLS
5172 .. 5192  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5172 .. 5192  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP160
5226 .. 5609  =  384 bp
128 amino acids  =  14.0 kDa
Product: 10 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)
VP160
5226 .. 5609  =  384 bp
128 amino acids  =  14.0 kDa
Product: 10 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)
T2A
5634 .. 5687  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
T2A
5634 .. 5687  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
PuroR
5697 .. 6296  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
5697 .. 6296  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
NeoR/KanR
7432 .. 8226  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
7432 .. 8226  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
6764 .. 7351  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6764 .. 7351  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
chimeric intron
813 .. 945  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
chimeric intron
813 .. 945  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
SV40 poly(A) signal
6485 .. 6566  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6485 .. 6566  =  82 bp
SV40 polyadenylation signal
attB1
960 .. 984  =  25 bp
recombination site for the Gateway® BP reaction
attB1
960 .. 984  =  25 bp
recombination site for the Gateway® BP reaction
attB2
6312 .. 6336  =  25 bp
recombination site for the Gateway® BP reaction
attB2
6312 .. 6336  =  25 bp
recombination site for the Gateway® BP reaction
ORF:  5392 .. 5655  =  264 bp
ORF:  87 amino acids  =  10.1 kDa
ORF:  7663 .. 7917  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  1005 .. 6296  =  5292 bp
ORF:  1763 amino acids  =  201.0 kDa
ORF:  7668 .. 8054  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2414 .. 2707  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  2978 .. 3427  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  5153 .. 5566  =  414 bp
ORF:  137 amino acids  =  14.2 kDa
ORF:  5119 .. 5658  =  540 bp
ORF:  179 amino acids  =  21.0 kDa
ORF:  7432 .. 8277  =  846 bp
ORF:  281 amino acids  =  30.9 kDa
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