pHR-SFFV-dCas9-BFP-KRAB

Lentiviral CRISPR interference vector for expressing catalytically inactive dCas9 fused to TagBFP and a KRAB domain.

Sequence Author: Qi

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SV40 NLS SV40 NLS pHR-SFFV-dCas9-BFP-KRAB 14,167 bp
dCas9
2801 .. 6904  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 1:  
   2801 .. 2827  =  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
2801 .. 6904  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 2:  
   2828 .. 2830  =  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
2801 .. 6904  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 3:  
   2831 .. 5317  =  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
2801 .. 6904  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 4:  
   5318 .. 5320  =  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
2801 .. 6904  =  4104 bp
1368 amino acids  =  158.3 kDa
5 segments
   Segment 5:  
   5321 .. 6904  =  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
2801 .. 6904  =  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
HA
6908 .. 6934  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
6908 .. 6934  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
SV40 NLS
6953 .. 6973  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6953 .. 6973  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6980 .. 7000  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6980 .. 7000  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
TagBFP
7040 .. 7735  =  696 bp
232 amino acids  =  26.2 kDa
Product: monomeric blue fluorescent protein
mammalian codon-optimized
TagBFP
7040 .. 7735  =  696 bp
232 amino acids  =  26.2 kDa
Product: monomeric blue fluorescent protein
mammalian codon-optimized
KRAB
7781 .. 7966  =  186 bp
62 amino acids  =  7.4 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
KRAB
7781 .. 7966  =  186 bp
62 amino acids  =  7.4 kDa
Product: Krüppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)
AmpR
9897 .. 10,757  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   9897 .. 9965  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9897 .. 10,757  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9966 .. 10,757  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9897 .. 10,757  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
LTR
1 .. 634  =  634 bp
long terminal repeat (LTR) from HIV-1
LTR
1 .. 634  =  634 bp
long terminal repeat (LTR) from HIV-1
WPRE
8030 .. 8618  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8030 .. 8618  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
10,928 .. 11,516  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
10,928 .. 11,516  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SFFV promoter
2319 .. 2726  =  408 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
SFFV promoter
2319 .. 2726  =  408 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
SV40 promoter
11,762 .. 12,091  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
11,762 .. 12,091  =  330 bp
SV40 enhancer and early promoter
LTR (ΔU3)
8728 .. 8961  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
LTR (ΔU3)
8728 .. 8961  =  234 bp
self-inactivating long terminal repeat (LTR) from HIV-1
RRE
1303 .. 1535  =  233 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1535  =  233 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
SV40 poly(A) signal
13,865 .. 13,999  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
13,865 .. 13,999  =  135 bp
SV40 polyadenylation signal
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
2049 .. 2166  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2049 .. 2166  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
9792 .. 9896  =  105 bp
AmpR promoter
9792 .. 9896  =  105 bp
SV40 ori
11,942 .. 12,077  =  136 bp
SV40 origin of replication
SV40 ori
11,942 .. 12,077  =  136 bp
SV40 origin of replication
ORF:  1642 .. 2241  =  600 bp
ORF:  199 amino acids  =  23.6 kDa
ORF:  13,369 .. 13,842  =  474 bp
ORF:  157 amino acids  =  18.2 kDa
ORF:  1181 .. 1519  =  339 bp
ORF:  112 amino acids  =  12.4 kDa
ORF:  2339 .. 2590  =  252 bp
ORF:  83 amino acids  =  9.2 kDa
ORF:  2801 .. 7969  =  5169 bp
ORF:  1722 amino acids  =  197.9 kDa
ORF:  8120 .. 8647  =  528 bp
ORF:  175 amino acids  =  19.3 kDa
ORF:  9194 .. 9541  =  348 bp
ORF:  115 amino acids  =  11.9 kDa
ORF:  12,374 .. 12,748  =  375 bp
ORF:  124 amino acids  =  14.0 kDa
ORF:  8133 .. 8807  =  675 bp
ORF:  224 amino acids  =  24.0 kDa
ORF:  9897 .. 10,757  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  12,528 .. 12,809  =  282 bp
ORF:  93 amino acids  =  10.7 kDa
ORF:  12,840 .. 13,223  =  384 bp
ORF:  127 amino acids  =  14.9 kDa
ORF:  2147 .. 2554  =  408 bp
ORF:  135 amino acids  =  15.2 kDa
ORF:  2675 .. 7543  =  4869 bp
ORF:  1622 amino acids  =  170.5 kDa
ORF:  9110 .. 9421  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  10,361 .. 10,627  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  12,422 .. 12,841  =  420 bp
ORF:  139 amino acids  =  15.9 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  4150 .. 4443  =  294 bp
ORF:  97 amino acids  =  10.7 kDa
ORF:  4714 .. 5163  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
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