SP-dCas9-VPR

Plasmid for mammalian cell expression of catalytically inactive Cas9m4 fused to the tripartite VP64-p65-Rta (VPR) transcriptional activator.

Sequence Author: Church Lab / Addgene #63798

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attB2 SV40 NLS SP-dCas9-VPR 11,319 bp
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 1:  
   748 .. 774  =  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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 2:  
   775 .. 777  =  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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 3:  
   778 .. 3261  =  2484 bp
   828 amino acids  =  96.1 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 4:  
   3262 .. 3267  =  6 bp
   2 amino acids  =  160.2 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 5:  
   3268 .. 3333  =  66 bp
   22 amino acids  =  2.5 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 6:  
   3334 .. 3336  =  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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 segments
   Segment 7:  
   3337 .. 4851  =  1515 bp
   505 amino acids  =  58.3 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
Cas9m4
748 .. 4851  =  4104 bp
1368 amino acids  =  158.2 kDa
7 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 mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)
SV40 NLS
4864 .. 4884  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4864 .. 4884  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP64
4975 .. 5124  =  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
4975 .. 5124  =  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)
SV40 NLS
5149 .. 5169  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
5149 .. 5169  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
RelA (p65) AD
5599 .. 5955  =  357 bp
119 amino acids  =  12.4 kDa
Product: transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)
RelA (p65) AD
5599 .. 5955  =  357 bp
119 amino acids  =  12.4 kDa
Product: transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)
Rta AD
5974 .. 6543  =  570 bp
190 amino acids  =  20.6 kDa
Product: transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)
Rta AD
5974 .. 6543  =  570 bp
190 amino acids  =  20.6 kDa
Product: transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)
AmpR
10,138 .. 10,998  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,138 .. 10,929  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,138 .. 10,998  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,930 .. 10,998  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,138 .. 10,998  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
7837 .. 8631  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
7837 .. 8631  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
9379 .. 9967  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9379 .. 9967  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6999 .. 7427  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6999 .. 7427  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
50 .. 429  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
50 .. 429  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
7441 .. 7770  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
7441 .. 7770  =  330 bp
SV40 enhancer and early promoter
CMV promoter
430 .. 633  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
430 .. 633  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
8807 .. 8928  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8807 .. 8928  =  122 bp
SV40 polyadenylation signal
AmpR promoter
10,999 .. 11,103  =  105 bp
AmpR promoter
10,999 .. 11,103  =  105 bp
HSV TK poly(A) signal
6749 .. 6797  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
6749 .. 6797  =  49 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
attB1
4921 .. 4945  =  25 bp
recombination site for the Gateway® BP reaction
attB1
4921 .. 4945  =  25 bp
recombination site for the Gateway® BP reaction
attB2
6548 .. 6572  =  25 bp
recombination site for the Gateway® BP reaction
attB2
6548 .. 6572  =  25 bp
recombination site for the Gateway® BP reaction
SV40 ori
7621 .. 7756  =  136 bp
SV40 origin of replication
SV40 ori
7621 .. 7756  =  136 bp
SV40 origin of replication
ORF:  589 .. 6546  =  5958 bp
ORF:  1985 amino acids  =  225.0 kDa
ORF:  7837 .. 8631  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5891 .. 6175  =  285 bp
ORF:  94 amino acids  =  10.0 kDa
ORF:  8009 .. 8395  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  10,268 .. 10,534  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  724 .. 1227  =  504 bp
ORF:  167 amino acids  =  18.5 kDa
ORF:  2152 .. 2670  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  2734 .. 3009  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  4261 .. 4794  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  8146 .. 8400  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  10,138 .. 10,998  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  648 .. 890  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  5529 .. 5762  =  234 bp
ORF:  77 amino acids  =  7.4 kDa
ORF:  6234 .. 6581  =  348 bp
ORF:  115 amino acids  =  12.7 kDa
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