Resources
Plasmid Files

pSPgRNA

Plasmid for expressing a Streptococcus pyogenes guide RNA in mammalian cells.

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pSPgRNA Sequence and MappSPgRNA.dna
Map and Sequence File   
Sequence Author:  Gersbach Lab / Addgene #47108
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 BbsI (903) BbsI (925) polIII terminator pSPgRNA 3223 bp
BbsI  (903)
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  (925)
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
2235 .. 3095  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2235 .. 3026  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2235 .. 3095  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3027 .. 3095  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2235 .. 3095  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1476 .. 2064  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1476 .. 2064  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3 .. 458  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3 .. 458  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
U6 promoter
659 .. 899  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
659 .. 899  =  241 bp
RNA polymerase III promoter for human U6 snRNA
AmpR promoter
3096 .. 3200  =  105 bp
AmpR promoter
3096 .. 3200  =  105 bp
gRNA scaffold
926 .. 1001  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
926 .. 1001  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
polIII terminator
1002 .. 1007  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
1002 .. 1007  =  6 bp
RNA polymerase III transcription terminator
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