pDD162 (Peft-3_Cas9 + Empty sgRNA)

C. elegans plasmid for expressing nematode codon-optimized Cas9 together with a single guide RNA (sgRNA).

Sequence Author: Goldstein Lab / Addgene #47549

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SV40 NLS pDD162 (Peft-3::Cas9 + Empty sgRNA) 8113 bp
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
   Segment 1:  
   2241 .. 3284  =  1044 bp
   348 amino acids  =  39.8 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
   Segment 2:  
   3336 .. 4406  =  1071 bp
   357 amino acids  =  42.2 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
   Segment 3:  
   4458 .. 5483  =  1026 bp
   342 amino acids  =  39.9 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
   Segment 4:  
   5535 .. 6497  =  963 bp
   321 amino acids  =  36.6 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
2241 .. 6497  =  4257 bp
1368 amino acids  =  158.4 kDa
4 segments  =  4104 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
SV40 NLS
6513 .. 6533  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6513 .. 6533  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
HA
6534 .. 6560  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
6534 .. 6560  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
AmpR
7916 .. 663  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7916 .. 7984  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7916 .. 663  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7985 .. 663  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7916 .. 663  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
eft-3 promoter
1625 .. 2224  =  600 bp
promoter for the C. elegans elongation factor 1α gene (Mitrovich and Anderson, 2000)
eft-3 promoter
1625 .. 2224  =  600 bp
promoter for the C. elegans elongation factor 1α gene (Mitrovich and Anderson, 2000)
ori
834 .. 1422  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
834 .. 1422  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CeU6 promoter
6967 .. 7316  =  350 bp
promoter for the C. elegans R07E5.16 U6 snRNA gene (Dickinson et al., 2013)
CeU6 promoter
6967 .. 7316  =  350 bp
promoter for the C. elegans R07E5.16 U6 snRNA gene (Dickinson et al., 2013)
tbb-2 terminator
6575 .. 6898  =  324 bp
transcription terminator for the C. elegans tubulin beta-2 gene
tbb-2 terminator
6575 .. 6898  =  324 bp
transcription terminator for the C. elegans tubulin beta-2 gene
AmpR promoter
7811 .. 7915  =  105 bp
AmpR promoter
7811 .. 7915  =  105 bp
gRNA scaffold
7318 .. 7393  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
7318 .. 7393  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
attB4
1604 .. 1624  =  21 bp
core recombination site for the Gateway® BP reaction
attB4
1604 .. 1624  =  21 bp
core recombination site for the Gateway® BP reaction
attB3
6902 .. 6922  =  21 bp
core recombination site for the Gateway® BP reaction
attB3
6902 .. 6922  =  21 bp
core recombination site for the Gateway® BP reaction
ORF:  7916 .. 663  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2241 .. 5516  =  3276 bp
ORF:  1091 amino acids  =  126.9 kDa
ORF:  5658 .. 6563  =  906 bp
ORF:  301 amino acids  =  34.3 kDa
ORF:  267 .. 533  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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