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Plasmid Files

PU6::klp-12_sgRNA

C. elegans plasmid for expressing a single guide RNA (sgRNA). Use together with Peft‑3::cas9-SV40_NLS::tbb-2 3'UTR.

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PU6_klp-12_sgRNA Sequence and MapPU6_klp-12_sgRNA.dna
Map and Sequence File   
Sequence Author:  Calarco Lab / Addgene #46170
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 EcoRI (396) polIII terminator HindIII (1242) PU6::klp-12_sgRNA 3481 bp
EcoRI  (396)
1 site
G A A T T C C T T A A G
HindIII  (1242)
1 site
A A G C T T T T C G A A
AmpR
2421 .. 3281  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2421 .. 3212  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2421 .. 3281  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3213 .. 3281  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2421 .. 3281  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1662 .. 2250  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1662 .. 2250  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CeU6 promoter
402 .. 901  =  500 bp
promoter for the C. elegans K09B11.12 U6 snRNA
gene (Friedland et al., 2013)
CeU6 promoter
402 .. 901  =  500 bp
promoter for the C. elegans K09B11.12 U6 snRNA
gene (Friedland et al., 2013)
3' CeU6
1005 .. 1241  =  237 bp
region downstream of the C. elegans K09B11.12 U6
snRNA gene
3' CeU6
1005 .. 1241  =  237 bp
region downstream of the C. elegans K09B11.12 U6
snRNA gene
AmpR promoter
3282 .. 3386  =  105 bp
AmpR promoter
3282 .. 3386  =  105 bp
gRNA scaffold
922 .. 997  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
922 .. 997  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
polIII terminator
998 .. 1003  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
998 .. 1003  =  6 bp
RNA polymerase III transcription terminator
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