pHAGE EF1-alpha dCas9-KRAB

Lentiviral CRISPR interference vector for constitutive expression of catalytically inactive dCas9 fused to a KRAB transcriptional repressor domain.

Sequence Author: Maehr

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pHAGE EF1 α dCas9-KRAB 12,843 bp
dCas9
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   8335 .. 8361  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   8362 .. 8364  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   8365 .. 10,851  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   10,852 .. 10,854  =  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
8335 .. 12,438  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   10,855 .. 12,438  =  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
8335 .. 12,438  =  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
12,451 .. 12,471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
12,451 .. 12,471  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
3xHA
12,472 .. 12,561  =  90 bp
30 amino acids  =  3.5 kDa
Product: three tandem HA epitope tags
3xHA
12,472 .. 12,561  =  90 bp
30 amino acids  =  3.5 kDa
Product: three tandem HA epitope tags
KRAB
12,598 .. 12,792  =  195 bp
65 amino acids  =  7.8 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
KRAB
12,598 .. 12,792  =  195 bp
65 amino acids  =  7.8 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
EF-1α promoter
7088 .. 8266  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
7088 .. 8266  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
2464 .. 3324  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   2464 .. 2532  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2464 .. 3324  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2533 .. 3324  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2464 .. 3324  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
LTR
4869 .. 5502  =  634 bp
long terminal repeat (LTR) from HIV-1
LTR
4869 .. 5502  =  634 bp
long terminal repeat (LTR) from HIV-1
PuroR
646 .. 1245  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
646 .. 1245  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
1258 .. 1846  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
1258 .. 1846  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
3495 .. 4083  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3495 .. 4083  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
126 .. 625  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
126 .. 625  =  500 bp
mouse phosphoglycerate kinase 1 promoter
LTR (ΔU3)
1921 .. 2154  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
LTR (ΔU3)
1921 .. 2154  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
RRE
6171 .. 6404  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
6171 .. 6404  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
HIV-1 Ψ
5549 .. 5674  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
5549 .. 5674  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
6926 .. 7043  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
6926 .. 7043  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
2359 .. 2463  =  105 bp
AmpR promoter
2359 .. 2463  =  105 bp
HA
67 .. 93  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
67 .. 93  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
attB1
8301 .. 8325  =  25 bp
recombination site for the Gateway® BP reaction
attB1
8301 .. 8325  =  25 bp
recombination site for the Gateway® BP reaction
attB2
12,833 .. 14  =  25 bp
recombination site for the Gateway® BP reaction
attB2
12,833 .. 14  =  25 bp
recombination site for the Gateway® BP reaction
EF-1α intron A
7319 .. 8257  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
7319 .. 8257  =  939 bp
intron upstream of the start codon of human EF-1α
ORF:  406 .. 1245  =  840 bp
ORF:  279 amino acids  =  30.1 kDa
ORF:  1348 .. 1860  =  513 bp
ORF:  170 amino acids  =  18.6 kDa
ORF:  2464 .. 3324  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6049 .. 6975  =  927 bp
ORF:  308 amino acids  =  35.4 kDa
ORF:  7723 .. 7956  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  8335 .. 12,795  =  4461 bp
ORF:  1486 amino acids  =  171.9 kDa
ORF:  1361 .. 1894  =  534 bp
ORF:  177 amino acids  =  18.8 kDa
ORF:  12,401 .. 12,739  =  339 bp
ORF:  112 amino acids  =  13.2 kDa
ORF:  6906 .. 7196  =  291 bp
ORF:  96 amino acids  =  10.5 kDa
ORF:  7644 .. 8081  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  8254 .. 8745  =  492 bp
ORF:  163 amino acids  =  17.9 kDa
ORF:  9739 .. 10,257  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  10,321 .. 10,596  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  11,224 .. 12,381  =  1158 bp
ORF:  385 amino acids  =  43.7 kDa
ORF:  2928 .. 3194  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5883 .. 6344  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  7386 .. 8006  =  621 bp
ORF:  206 amino acids  =  23.0 kDa
ORF:  6749 .. 6997  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
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Download pHAGE EF1-alpha dCas9-KRAB.dna file

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