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Plasmid Files

pHsp70-Cas9

Plasmid for expressing S. pyogenes Cas9 in Drosophila cells from the hsp70 promoter. Use together with pU6‑BbsI-chiRNA.

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pHsp70-Cas9 Sequence and MappHsp70-Cas9.dna
Map and Sequence File   
Sequence Author:  O'Connor-Giles Lab / Addgene #45945
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 NotI (6560) NotI (819) ATG SV40 NLS pHsp70-Cas9 8149 bp
NotI  (6560)
2 sites
G C G G C C G C C G C C G G C G
NotI  (819)
2 sites
G C G G C C G C C G C C G G C G
Cas9
1429 .. 5529  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
1429 .. 5529  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
hsp70 poly(A)
5657 .. 6529  =  873 bp
polyadenlyation signal from the Drosophila
melanogaster
hsp70Ab gene
hsp70 poly(A)
5657 .. 6529  =  873 bp
polyadenlyation signal from the Drosophila
melanogaster
hsp70Ab gene
NeoR/KanR
6742 .. 7536  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
6742 .. 7536  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
182 .. 770  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
182 .. 770  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hsp70 promoter
841 .. 1293  =  453 bp
Drosophila melanogaster hsp70Bb promoter
hsp70 promoter
841 .. 1293  =  453 bp
Drosophila melanogaster hsp70Bb promoter
3xFLAG
1312 .. 1377  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
1312 .. 1377  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
nucleoplasmin NLS
5530 .. 5577  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
5530 .. 5577  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
SV40 NLS
1384 .. 1404  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
1384 .. 1404  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
ATG
1309 .. 1311  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1309 .. 1311  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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