pNA0304
(Plasmid #165612)

• Purpose: Vector for expression of sgRNAs in yeast: SNR52p-NotI-sgRNA-SUP4t (NotI site in place of the spacer)
• Depositing Lab: Marcus Noyes
• Publication: Goldberg et al Nat Commun. 2021 Jan 13;12(1):349. doi: 10.1038/s41467-020-20650-x.

Backbone

• Vector backbone: p426-SNR52p-gRNA.CAN1.Y-SUP4t
• Backbone manufacturer: Church Lab (Addgene #43803)
• Vector type: Yeast Expression, CRISPR
• Selectable markers: URA3

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SNR52p-NotI-sgRNA-SUP4t (S. cerevisiae SNR52 promoter driving the sgRNA for SpCas9, with a NotI site in place of the spacer)
• Mutation: NotI site replaced the CAN1 spacer in parent vector (to facilitate insertion of new spacers via Gibson assembly)
• Promoter: SNR52

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: M13R
• 3′ sequencing primer: M13F

Resource Information

• Supplemental Documents:
  • pNA0304_p426-SNR52p-gRNA.NotI-SUP4t.gbk

Terms and Licenses

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

Depositor Comments

This plasmid was constructed by Dr. Agmon in the Boeke lab and first described in:
Agmon, N. et al. Phylogenetic debugging of a complete human biosynthetic pathway transplanted into yeast. Nucleic Acids Res. 48, 486–499 (2020)