pCT-tRNA
(Plasmid #133813)

• Purpose: CRISPR-Cas9 system for genetic manipulation of Candida tropicalis
• Depositing Lab: Geraldine Butler
• Publication: Lombardi et al mSphere. 2019 Mar 13;4(2). pii: 4/2/e00125-19. doi: 10.1128/mSphere.00125-19.

Backbone

• Vector backbone: pAYCU268
• Vector type: CRISPR
• Selectable markers: SAT1 (resistance to Nourseothricin)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: cassette for the expression of the sgRNA from the Ashbya gossypii RNA pol II TEF1 promoter
• gRNA/shRNA sequence: Unknown
• Species: Synthetic

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: Unknown

Resource Information

• Supplemental Documents:
  • pCT-tRNA.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The plasmid contains the following relevant inserts: 1. C. albicans Autonomously Replicating Sequence 2 for episomal propagation in C. tropicalis; 2. CAS9 sequence expressed from a Meyerozyma guilliermondii promoter; 3. Cassette for the expression of the sgRNA from a Ashbya gossypii RNA pol II promoter. The mature sgRNA is released following RNase Z processing of the upstream tRNA and self-cleavage of the dowstream HDV Ribozyme. The guide targeting the gene of interest can be cloned into the SapI/BspQI digested plasmid. The regulatory elements (species of origin of promoters/terminators) are described in the map.