Resources
Plasmid Files

pT7-gRNA

Plasmid for in vitro expression of a single guide RNA (sgRNA) using the T7 promoter.

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pT7-gRNA Sequence and MappT7-gRNA.dna
Map and Sequence File   
Sequence Author:  Chen Lab / Addgene #46759
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 M13 fwd BsmBI (311) BsmBI (348) pT7-gRNA 2542 bp
BsmBI  (311)
2 sites
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI  (348)
2 sites
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
AmpR
1552 .. 2412  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1552 .. 2343  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1552 .. 2412  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2344 .. 2412  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1552 .. 2412  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
793 .. 1381  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
793 .. 1381  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
2413 .. 2517  =  105 bp
AmpR promoter
2413 .. 2517  =  105 bp
gRNA scaffold
240 .. 315  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
240 .. 315  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
T7 promoter
349 .. 367  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
349 .. 367  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
196 .. 212  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
196 .. 212  =  17 bp
common sequencing primer, one of multiple similar
variants
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