Resources
Plasmid Files

pU6-BbsI-chiRNA

Plasmid for expressing a chimeric RNA in Drosophila cells from the U6 promoter. Use together with pHsp70‑Cas9.

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pU6-BbsI-chiRNA.dna
Map and Sequence File:    Download    Open   
Sequence Author:  O'Connor-Giles Lab / Addgene #45946
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BbsI (1093) BbsI (1115) pU6-BbsI-chiRNA 3458 bp
BbsI  (1093)
2 sites
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (1115)
2 sites
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
AmpR
2473 .. 3333  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2473 .. 3264  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2473 .. 3333  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3265 .. 3333  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2473 .. 3333  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1714 .. 2302  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1714 .. 2302  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
7 .. 462  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
7 .. 462  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
dU6 promoter
698 .. 1097  =  400 bp
RNA polymerase III promoter for Drosophila U6-2 snRNA (Das et al., 1987)
dU6 promoter
698 .. 1097  =  400 bp
RNA polymerase III promoter for Drosophila U6-2 snRNA (Das et al., 1987)
AmpR promoter
3334 .. 3438  =  105 bp
AmpR promoter
3334 .. 3438  =  105 bp
gRNA scaffold
1116 .. 1191  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
1116 .. 1191  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
polIII terminator
1192 .. 1197  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
1192 .. 1197  =  6 bp
RNA polymerase III transcription terminator
ORF:  2603 .. 2869  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  474 .. 767  =  294 bp
ORF:  97 amino acids  =  11.1 kDa
ORF:  2473 .. 3333  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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