• PURPOSE

  Plasmid encoding iCas9, tracrRNA and crRNAs

BACKBONE

• Vector backbone
  pRS426
• Backbone size w/o insert (bp)5000
• Total vector size (bp)10805
• Modifications to backbone
  URA3 promoter was truncated
• Vector type
  Yeast Expression, CRISPR
• Selectable markers
  URA3

• Gene/Insert name
  iCas9
• Species
  Streptococcus pyogenes
• Insert Size (bp)
  4179
• Mutation
  changed Aspartate 147 to Tyrosine, Proline 411 to Threonine
• PromoterTEF1p
• Tags / Fusion Proteins
  • FLAG (N terminal on insert)
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)

Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step Multigene Disruption in Saccharomyces cerevisiae. Bao Z, Xiao H, Liang J, Zhang L, Xiong X, Sun N, Si T, Zhao H. ACS Synth Biol. 2014 Sep 19. 10.1021/sb500255k PubMed 25207793