Plasmids
- Plasmids carrying biosynthetic gene clusters for heterologous expression in S. cerevisiae, described in Harvey CJB et al., bioRxiv (2018), will soon be available from public repositories and in the meantime are available upon request from the corresponding authors. Download their sequences here.
- pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNA: Plasmid for inducible and tunable CRISPR interference in S. cerevisiae. pRS416 Ura marked Cen/Ars plasmid with dCas9-Mxi1 under Tef1 promoter, and tet-incucible RPR1 promoter with NotI cloning site adjacent to gRNA. Available from AddGene.
- pWEB436: integrative E. coli-Streptomyces shuttle vector
- pTARa: a BAC-based S. cerevisiae/E. coli/Streptomyces shuttle capture vector
- pJWC1: an IncP1-α group broad-host-range cosmid vector
Strains
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Our CRISPRi collection contains ~9000 S. cerevisiae strains representing 1600 genes (~1100 genes essential for fermentative growth and ~500 genes required for respiration). Each archived strain constitutely expresses dCas9-Mxi1 and contains an integrated, sequence-verified gRNA under control of ATc-inducible promoter. Ref: Smith JS et al., Mol Sys Biol (2017). Please contact Bob St. Onge for access.
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Optimized S. cerevisiae strains for heterologous expression of natural product biosynthetic gene clusters. These strains are derived from BY4741 (S288c), and several genetic defects have been repaired to improve sporulation efficiency and mitochondrial genome stability, which allow robust induction of biosynthetic gene expression during respiratory growth. See the genotype table for details. Ref: Harvey CJB et al., bioRxiv (2018). Please contact Angela Chu for access.
