Resources
Plasmid Files

gRNA_Cloning Vector

Mammalian cell plasmid for generating a guide RNA (gRNA) expression cassette by oligo synthesis and Gibson assembly.

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gRNA_Cloning Vector.dna
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Sequence Author:  Church Lab / Addgene #41824
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AflII (431) gRNA_Cloning Vector 3914 bp
AflII  (431)
1 site
C T T A A G G A A T T C
NeoR/KanR
1633 .. 2427  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1633 .. 2427  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3146 .. 3733  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3146 .. 3733  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
BleoR
2634 .. 3008  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
2634 .. 3008  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
ccdB
982 .. 1284  =  303 bp
100 amino acids  =  11.6 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
ccdB
982 .. 1284  =  303 bp
100 amino acids  =  11.6 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
U6 promoter (truncated)
434 .. 663  =  230 bp
RNA polymerase III promoter for human U6 snRNA, missing the GTGGAAAGGAC sequence at the 3' end
U6 promoter (truncated)
434 .. 663  =  230 bp
RNA polymerase III promoter for human U6 snRNA, missing the GTGGAAAGGAC sequence at the 3' end
gRNA scaffold (3' portion)
378 .. 433  =  56 bp
3' portion of the guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold (3' portion)
378 .. 433  =  56 bp
3' portion of the guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
polIII terminator
372 .. 377  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
372 .. 377  =  6 bp
RNA polymerase III transcription terminator
ORF:  1633 .. 2427  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  779 .. 1078  =  300 bp
ORF:  99 amino acids  =  11.1 kDa
ORF:  1805 .. 2191  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2634 .. 3008  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  2606 .. 2914  =  309 bp
ORF:  102 amino acids  =  10.9 kDa
ORF:  1942 .. 2475  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
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