dCAS9-VP64_GFP

Zhang lab lentiviral vector encoding enhanced GFP (EGFP) plus catalytically inactive dCas9 fused to the VP64 transcriptional activation domain.

Sequence Author: Zhang Lab / Addgene #61422

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AmpR promoter SV40 NLS HIV-1 Ψ ATG dCAS9-VP64_GFP 14,382 bp
ATG
3872 .. 3874  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3872 .. 3874  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
3878 .. 3898  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
3878 .. 3898  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
dCas9
3908 .. 8008  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   3908 .. 3931  =  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
3908 .. 8008  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   3932 .. 3934  =  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
3908 .. 8008  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   3935 .. 6421  =  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
3908 .. 8008  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   6422 .. 6424  =  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
3908 .. 8008  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   6425 .. 8008  =  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
3908 .. 8008  =  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
8063 .. 8083  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
8063 .. 8083  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP64
8108 .. 8257  =  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
8108 .. 8257  =  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)
T2A
8279 .. 8332  =  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
8279 .. 8332  =  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.
EGFP
8333 .. 9049  =  717 bp
238 amino acids  =  26.8 kDa
2 segments
   Segment 1:  1a  
   8333 .. 8335  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
8333 .. 9049  =  717 bp
238 amino acids  =  26.8 kDa
2 segments
   Segment 2:  
   8336 .. 9049  =  714 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
8333 .. 9049  =  717 bp
238 amino acids  =  26.8 kDa
2 segments
Product: enhanced GFP
mammalian codon-optimized
EF-1α promoter
2682 .. 3859  =  1178 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
2682 .. 3859  =  1178 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
13,389 .. 14,249  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   13,389 .. 14,180  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
13,389 .. 14,249  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   14,181 .. 14,249  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
13,389 .. 14,249  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
9074 .. 9662  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
9074 .. 9662  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
12,630 .. 13,218  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
12,630 .. 13,218  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
10,669 .. 11,097  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
10,669 .. 11,097  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
238 .. 617  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
238 .. 617  =  380 bp
human cytomegalovirus immediate early enhancer
BleoR
11,554 .. 11,928  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
11,554 .. 11,928  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
11,111 .. 11,440  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
11,111 .. 11,440  =  330 bp
SV40 enhancer and early promoter
RRE
1680 .. 1913  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1680 .. 1913  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
bGH poly(A) signal
10,399 .. 10,623  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
10,399 .. 10,623  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
LTR (truncated)
835 .. 1015  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
835 .. 1015  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
10,187 .. 10,367  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
10,187 .. 10,367  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1062 .. 1187  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
1062 .. 1187  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
12,058 .. 12,179  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
12,058 .. 12,179  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2440 .. 2557  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2440 .. 2557  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
14,250 .. 14,354  =  105 bp
AmpR promoter
14,250 .. 14,354  =  105 bp
EM7 promoter
11,488 .. 11,535  =  48 bp
synthetic bacterial promoter
EM7 promoter
11,488 .. 11,535  =  48 bp
synthetic bacterial promoter
EF-1α intron A
2913 .. 3850  =  938 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
2913 .. 3850  =  938 bp
intron upstream of the start codon of human EF-1α
SV40 ori
11,291 .. 11,426  =  136 bp
SV40 origin of replication
SV40 ori
11,291 .. 11,426  =  136 bp
SV40 origin of replication
ORF:  1558 .. 2445  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3316 .. 3549  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  11,554 .. 11,928  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  13,519 .. 13,785  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2420 .. 2803  =  384 bp
ORF:  127 amino acids  =  14.5 kDa
ORF:  3872 .. 9049  =  5178 bp
ORF:  1725 amino acids  =  196.9 kDa
ORF:  9164 .. 10,213  =  1050 bp
ORF:  349 amino acids  =  39.4 kDa
ORF:  3237 .. 3674  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  9177 .. 9689  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  10,620 .. 10,886  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  5254 .. 5547  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  5818 .. 6267  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  7993 .. 8373  =  381 bp
ORF:  126 amino acids  =  12.9 kDa
ORF:  1473 .. 1853  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  2598 .. 3599  =  1002 bp
ORF:  333 amino acids  =  36.1 kDa
ORF:  11,547 .. 11,834  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  13,389 .. 14,249  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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