pMB66

C. elegans plasmid for expression of nematode codon-optimized Cas9 fused to monomeric EGFP and nuclear localization signals.

Sequence Author: Boxem Lab / Addgene #47946

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ATG SV40 NLS pMB66 9000 bp
ATG
1045 .. 1047  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1045 .. 1047  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
1048 .. 1113  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
1048 .. 1113  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
1120 .. 1140  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1120 .. 1140  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
   Segment 1:  
   1165 .. 2205  =  1041 bp
   347 amino acids  =  39.7 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
   Segment 2:  
   2257 .. 3243  =  987 bp
   329 amino acids  =  38.9 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
   Segment 3:  
   3295 .. 4356  =  1062 bp
   354 amino acids  =  41.2 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
   Segment 4:  
   4408 .. 5418  =  1011 bp
   337 amino acids  =  38.5 kDa
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
Cas9
1165 .. 5418  =  4254 bp
1367 amino acids  =  158.3 kDa
4 segments  =  4101 bp
Product: S. pyogenes Cas9, codon optimized for C. elegans with synthetic introns
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
   Segment 1:  
   5425 .. 5727  =  303 bp
   101 amino acids  =  11.4 kDa
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
   Segment 2:  
   5781 .. 5855  =  75 bp
   25 amino acids  =  2.9 kDa
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
   Segment 3:  
   5907 .. 6026  =  120 bp
   40 amino acids  =  4.7 kDa
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
   Segment 4:  
   6078 .. 6293  =  216 bp
   72 amino acids  =  8.1 kDa
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
mEGFP
5425 .. 6293  =  869 bp
238 amino acids  =  26.9 kDa
4 segments  =  714 bp
Product: monomeric enhanced GFP, optimized with synthetic introns for expression in C. elegans
EGL-13 NLS
6300 .. 6374  =  75 bp
25 amino acids  =  2.9 kDa
Product: nuclear localization signal from the C. elegans EGL-13 transcription factor (Lyssenko et al., 2007)
EGL-13 NLS
6300 .. 6374  =  75 bp
25 amino acids  =  2.9 kDa
Product: nuclear localization signal from the C. elegans EGL-13 transcription factor (Lyssenko et al., 2007)
AmpR
8015 .. 8875  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8015 .. 8806  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8015 .. 8875  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8807 .. 8875  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8015 .. 8875  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7256 .. 7844  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7256 .. 7844  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6 .. 461  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6 .. 461  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
tbb-2 terminator
6384 .. 6707  =  324 bp
transcription terminator for the C. elegans tubulin beta-2 gene
tbb-2 terminator
6384 .. 6707  =  324 bp
transcription terminator for the C. elegans tubulin beta-2 gene
hsp16-48 promoter
730 .. 1034  =  305 bp
promoter for the C. elegans heat shock protein 16-48 gene (Russnak and Candido, 1985)
hsp16-48 promoter
730 .. 1034  =  305 bp
promoter for the C. elegans heat shock protein 16-48 gene (Russnak and Candido, 1985)
AmpR promoter
8876 .. 8980  =  105 bp
AmpR promoter
8876 .. 8980  =  105 bp
ORF:  1045 .. 4389  =  3345 bp
ORF:  1114 amino acids  =  129.4 kDa
ORF:  4441 .. 5748  =  1308 bp
ORF:  435 amino acids  =  49.5 kDa
ORF:  8145 .. 8411  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  244 .. 687  =  444 bp
ORF:  147 amino acids  =  16.3 kDa
ORF:  474 .. 914  =  441 bp
ORF:  146 amino acids  =  16.3 kDa
ORF:  3291 .. 3515  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  4428 .. 4673  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
ORF:  6066 .. 6404  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  8015 .. 8875  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pMB66.dna file

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