pMLM3705

CRISPR activation plasmid for mammalian cell expression of catalytically inactive dCas9 fused to the VP64 transcriptional activation domain.

Sequence Author: Joung Lab / Addgene #47754

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pMLM3705 7785 bp
dCas9
409 .. 4512  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   409 .. 435  =  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
409 .. 4512  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   436 .. 438  =  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
409 .. 4512  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   439 .. 2925  =  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
409 .. 4512  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   2926 .. 2928  =  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
409 .. 4512  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   2929 .. 4512  =  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
409 .. 4512  =  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
4519 .. 4539  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4519 .. 4539  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
3xFLAG
4546 .. 4611  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
4546 .. 4611  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
VP64
4642 .. 4791  =  150 bp
50 amino acids  =  5.5 kDa
Product: tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)
VP64
4642 .. 4791  =  150 bp
50 amino acids  =  5.5 kDa
Product: tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)
AmpR
6302 .. 7162  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6302 .. 7093  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6302 .. 7162  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7094 .. 7162  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6302 .. 7162  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
5543 .. 6131  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5543 .. 6131  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
7537 .. 131  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
7537 .. 131  =  380 bp
human cytomegalovirus immediate early enhancer
bGH poly(A) signal
4846 .. 5070  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
4846 .. 5070  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
132 .. 335  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
132 .. 335  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AmpR promoter
7163 .. 7267  =  105 bp
AmpR promoter
7163 .. 7267  =  105 bp
T7 promoter
377 .. 395  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
377 .. 395  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  409 .. 4794  =  4386 bp
ORF:  1461 amino acids  =  168.5 kDa
ORF:  4559 .. 4858  =  300 bp
ORF:  99 amino acids  =  11.2 kDa
ORF:  6432 .. 6698  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2373 .. 2660  =  288 bp
ORF:  95 amino acids  =  10.5 kDa
ORF:  3522 .. 3767  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
ORF:  4590 .. 4817  =  228 bp
ORF:  75 amino acids  =  7.8 kDa
ORF:  4463 .. 4726  =  264 bp
ORF:  87 amino acids  =  10.4 kDa
ORF:  6302 .. 7162  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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