pHAGE-TO-dCas9-3XGFP

Lentiviral vector for labeling chromosomal loci by tetracycline-regulatable expression of catalytically inactive dCas9 fused to three copies of GFP.

Sequence Author: Pederson Lab / Addgene #64107

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CMV promoter tet operator tet operator ATG pHAGE-TO-dCas9-3XGFP 12,986 bp
ATG
2980 .. 2982  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2980 .. 2982  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
2986 .. 3006  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2986 .. 3006  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
dCas9
3025 .. 7125  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 1:  
   3025 .. 3048  =  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
3025 .. 7125  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 2:  
   3049 .. 3051  =  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
3025 .. 7125  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 3:  
   3052 .. 5538  =  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
3025 .. 7125  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 4:  
   5539 .. 5541  =  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
3025 .. 7125  =  4101 bp
1367 amino acids  =  158.2 kDa
5 segments
   Segment 5:  
   5542 .. 7125  =  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
3025 .. 7125  =  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
7144 .. 7164  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
7144 .. 7164  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
superfolder GFP
7177 .. 7890  =  714 bp
238 amino acids  =  26.8 kDa
Product: GFP variant that folds robustly even when fused to poorly folded proteins (Pédelacq et al., 2006)
superfolder GFP
7177 .. 7890  =  714 bp
238 amino acids  =  26.8 kDa
Product: GFP variant that folds robustly even when fused to poorly folded proteins (Pédelacq et al., 2006)
superfolder GFP
7909 .. 8622  =  714 bp
238 amino acids  =  26.8 kDa
Product: GFP variant that folds robustly even when fused to poorly folded proteins (Pédelacq et al., 2006)
superfolder GFP
7909 .. 8622  =  714 bp
238 amino acids  =  26.8 kDa
Product: GFP variant that folds robustly even when fused to poorly folded proteins (Pédelacq et al., 2006)
superfolder GFP
8641 .. 9354  =  714 bp
238 amino acids  =  26.8 kDa
Product: GFP variant that folds robustly even when fused to poorly folded proteins (Pédelacq et al., 2006)
superfolder GFP
8641 .. 9354  =  714 bp
238 amino acids  =  26.8 kDa
Product: GFP variant that folds robustly even when fused to poorly folded proteins (Pédelacq et al., 2006)
AmpR
10,582 .. 11,442  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   10,582 .. 10,650  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,582 .. 11,442  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   10,651 .. 11,442  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,582 .. 11,442  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
9376 .. 9964  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
9376 .. 9964  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
11,613 .. 12,201  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
11,613 .. 12,201  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
2299 .. 2602  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2299 .. 2602  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
3' LTR (ΔU3)
10,039 .. 10,272  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
10,039 .. 10,272  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
CMV promoter
2603 .. 2806  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2603 .. 2806  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2058 .. 2175  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2058 .. 2175  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
10,477 .. 10,581  =  105 bp
AmpR promoter
10,477 .. 10,581  =  105 bp
tet operator
2808 .. 2826  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2808 .. 2826  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2829 .. 2847  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2829 .. 2847  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  2980 .. 9363  =  6384 bp
ORF:  2127 amino acids  =  243.1 kDa
ORF:  9466 .. 9978  =  513 bp
ORF:  170 amino acids  =  18.6 kDa
ORF:  10,582 .. 11,442  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1181 .. 2107  =  927 bp
ORF:  308 amino acids  =  35.4 kDa
ORF:  2762 .. 3151  =  390 bp
ORF:  129 amino acids  =  14.8 kDa
ORF:  9479 .. 10,012  =  534 bp
ORF:  177 amino acids  =  18.8 kDa
ORF:  1881 .. 2129  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
ORF:  11,046 .. 11,312  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  2875 .. 3432  =  558 bp
ORF:  185 amino acids  =  20.6 kDa
ORF:  4426 .. 4944  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  5008 .. 5283  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  5911 .. 7068  =  1158 bp
ORF:  385 amino acids  =  43.7 kDa
ORF:  7072 .. 9333  =  2262 bp
ORF:  753 amino acids  =  87.4 kDa
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