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 Sequence and MapgRNA_Cloning Vector.dna
Map and Sequence File   
Sequence Author:  Church Lab / Addgene #41824
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 polIII terminator AflII (431) gRNA_Cloning Vector 3914 bp
AflII  (431)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
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
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