lenti dCAS-VP64_Blast

Zhang lab lentiviral vector encoding a blasticidin resistance marker plus catalytically inactive dCas9 fused to the VP64 transcriptional activation domain.

Sequence Author: Zhang Lab / Addgene #61425

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LTR (truncated) SV40 NLS CMV enhancer ATG lenti dCAS-VP64_Blast 14,085 bp
ATG
3878 .. 3880  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3878 .. 3880  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
nucleoplasmin NLS
3881 .. 3928  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
3881 .. 3928  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
dCas9
3929 .. 8029  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   3929 .. 3952  =  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 N863A mutation in the HNH catalytic domain
dCas9
3929 .. 8029  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   3953 .. 3955  =  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 N863A mutation in the HNH catalytic domain
dCas9
3929 .. 8029  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   3956 .. 6511  =  2556 bp
   852 amino acids  =  98.9 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 N863A mutation in the HNH catalytic domain
dCas9
3929 .. 8029  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   6512 .. 6514  =  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 N863A mutation in the HNH catalytic domain
dCas9
3929 .. 8029  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   6515 .. 8029  =  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 D10A mutation in the RuvC catalytic domain and the N863A mutation in the HNH catalytic domain
dCas9
3929 .. 8029  =  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 N863A mutation in the HNH catalytic domain
SV40 NLS
8084 .. 8104  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
8084 .. 8104  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
VP64
8129 .. 8278  =  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
8129 .. 8278  =  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
8300 .. 8353  =  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
8300 .. 8353  =  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.
BSD
8354 .. 8752  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
8354 .. 8752  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
EF-1α promoter
2687 .. 3865  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
2687 .. 3865  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
13,092 .. 13,952  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   13,092 .. 13,883  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
13,092 .. 13,952  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   13,884 .. 13,952  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
13,092 .. 13,952  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
8777 .. 9365  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8777 .. 9365  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
12,333 .. 12,921  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
12,333 .. 12,921  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
10,372 .. 10,800  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
10,372 .. 10,800  =  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
11,257 .. 11,631  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
11,257 .. 11,631  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
10,814 .. 11,143  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
10,814 .. 11,143  =  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
10,102 .. 10,326  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
10,102 .. 10,326  =  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)
9890 .. 10,070  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
9890 .. 10,070  =  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
11,761 .. 11,882  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
11,761 .. 11,882  =  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
13,953 .. 14,057  =  105 bp
AmpR promoter
13,953 .. 14,057  =  105 bp
EM7 promoter
11,191 .. 11,238  =  48 bp
synthetic bacterial promoter
EM7 promoter
11,191 .. 11,238  =  48 bp
synthetic bacterial promoter
EF-1α intron A
2918 .. 3856  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
2918 .. 3856  =  939 bp
intron upstream of the start codon of human EF-1α
SV40 ori
10,994 .. 11,129  =  136 bp
SV40 origin of replication
SV40 ori
10,994 .. 11,129  =  136 bp
SV40 origin of replication
ORF:  1558 .. 2445  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3322 .. 3555  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  11,257 .. 11,631  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  13,222 .. 13,488  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3878 .. 8752  =  4875 bp
ORF:  1624 amino acids  =  184.3 kDa
ORF:  8867 .. 9916  =  1050 bp
ORF:  349 amino acids  =  39.4 kDa
ORF:  3243 .. 3680  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  8880 .. 9392  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  10,323 .. 10,589  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  5275 .. 5568  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  5839 .. 6288  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  8014 .. 8274  =  261 bp
ORF:  86 amino acids  =  8.9 kDa
ORF:  1473 .. 1853  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  2985 .. 3605  =  621 bp
ORF:  206 amino acids  =  23.0 kDa
ORF:  11,250 .. 11,537  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  13,092 .. 13,952  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  8318 .. 8656  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
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