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Plasmid Files

pMZ374

Fission yeast plasmid for expressing S. pyogenes Cas9 from the adh1 promoter plus a single guide RNA (sgRNA) from the rrk1 promoter/leader.

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pMZ374 Sequence and MappMZ374.dna
Map and Sequence File   
Sequence Author:  Zaratiegui Lab / Addgene #59896
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 SV40 NLS sTRSV HHRz CspCI (3567) CspCI (3601) pMZ374 11,384 bp
CspCI  (3567)
2 sites
( N ) 10-11 C A A N N N N N G T G G ( N ) 10-11 N N N N ( N ) 10-11 G T T N N N N N C A C C ( N ) 10-11

Efficient cleavage requires at least two copies of the CspCI
recognition sequence.
CspCI makes two double-stranded cuts.
Sticky ends from different CspCI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
CspCI  (3601)
2 sites
( N ) 10-11 C A A N N N N N G T G G ( N ) 10-11 N N N N ( N ) 10-11 G T T N N N N N C A C C ( N ) 10-11

Efficient cleavage requires at least two copies of the CspCI
recognition sequence.
CspCI makes two double-stranded cuts.
Sticky ends from different CspCI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Cas9
6911 .. 11,014  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
6911 .. 11,014  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
AmpR
5155 .. 6015  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5155 .. 5946  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5155 .. 6015  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5947 .. 6015  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5155 .. 6015  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
S. pombe ura4
1930 .. 2724  =  795 bp
264 amino acids  =  29.4 kDa
Product: orotidine 5'-phosphate decarboxylase,
required for uracil biosynthesis
S. pombe ura4
1930 .. 2724  =  795 bp
264 amino acids  =  29.4 kDa
Product: orotidine 5'-phosphate decarboxylase,
required for uracil biosynthesis
ars1
539 .. 1320  =  782 bp
Schizosaccharomyces pombe autonomously
replicating sequence ars1
ars1
539 .. 1320  =  782 bp
Schizosaccharomyces pombe autonomously
replicating sequence ars1
adh1 promoter
6149 .. 6890  =  742 bp
promoter for the Schizosaccharomyces pombe adh1
gene encoding alcohol dehydrogenase
adh1 promoter
6149 .. 6890  =  742 bp
promoter for the Schizosaccharomyces pombe adh1
gene encoding alcohol dehydrogenase
ori
4396 .. 4984  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4396 .. 4984  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rrk1 promoter/leader
3601 .. 3957  =  357 bp
promoter and leader RNA for the
Schizosaccharomyces pombe rrk1 gene encoding
the K-RNA component of RNase P (Jacobs et al.,
2014)
rrk1 promoter/leader
3601 .. 3957  =  357 bp
promoter and leader RNA for the
Schizosaccharomyces pombe rrk1 gene encoding
the K-RNA component of RNase P (Jacobs et al.,
2014)
CYC1 terminator
11,063 .. 11,310  =  248 bp
transcription terminator for CYC1
CYC1 terminator
11,063 .. 11,310  =  248 bp
transcription terminator for CYC1
AmpR promoter
6016 .. 6120  =  105 bp
AmpR promoter
6016 .. 6120  =  105 bp
gRNA scaffold
3483 .. 3566  =  84 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
3483 .. 3566  =  84 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
sTRSV HHRz
3431 .. 3482  =  52 bp
hammerhead ribozyme from the tobacco ringspot
virus satellite RNA (Khvorova et al., 2003)
sTRSV HHRz
3431 .. 3482  =  52 bp
hammerhead ribozyme from the tobacco ringspot
virus satellite RNA (Khvorova et al., 2003)
SV40 NLS
11,027 .. 11,047  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
11,027 .. 11,047  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
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