pYZ038
(Plasmid #200227)

• Purpose: universal template to build tRNA-gRNA array for multiplex editing CREEPY
• Depositing Lab: Jef Boeke
• Publication: Zhao et al Nucleic Acids Res. 2023 Jun 16:gkad491. doi: 10.1093/nar/gkad491.

Backbone

• Vector backbone: pCR-Blunt II-TOPO
• Backbone manufacturer: Thermo Fisher
• Backbone size w/o insert (bp): 3500
• Total vector size (bp): 3673
• Vector type: Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: 30°C is recommended and 37°C is also fine
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: tRNA-gRNA scaffold
• Species: S. cerevisiae (budding yeast), Synthetic
• Insert Size (bp): 153

Resource Information

• Supplemental Documents:
  • pYZ038 Topo clone of tRNA-gRNA scaffold for Cas9.gb

Terms and Licenses

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

Depositor Comments

A common PCR template used fo CRISPR Engineering of EPisomes in Yeast, or CREEPY, a method for rapid engineering of large synthetic episomal DNA constructs. This is designed and assembled based on the template consisting of gRNA scaffold with a tRNA sequence, as published in Zhang, Yueping, et al. "A gRNA-tRNA array for CRISPR-Cas9 based rapid multiplexed genome editing in Saccharomyces cerevisiae." Nature communications 10.1 (2019): 1053.