3xFLAG-dCas9/pTEF1p-CYC1t
(Plasmid #62190)

• Purpose: Expresses 3xFLAG-dCas9 in budding yeast for enChIP analysis to purify specific genomic regions of interest.
• Depositing Lab: Hodaka Fujii
• Publication: Fujii et al Biol Methods Protoc. 2022 Oct 17;7(1):bpac025. doi: 10.1093/biomethods/bpac025. eCollection 2022.

Backbone

• Vector backbone: p414
• Backbone size w/o insert (bp): 5385
• Total vector size (bp): 9596
• Vector type: Yeast Expression, CRISPR
• Selectable markers: TRP1

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: 3xFLAG-dCas9
• Species: Synthetic; S. pyogenes
• Insert Size (bp): 4212
• Mutation: human codon-optimized, D10A, H840A
• Promoter: TEF1 promoter
• Tags / Fusion Proteins:
  • 3xFLAG (N terminal on insert)
  • NLS (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T3 promoter
• 3′ sequencing primer: T7 promoter

Resource Information

• A portion of this plasmid was derived from a plasmid made by: George Church (Addgene plasmid 43802)

Terms and Licenses

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

Depositor Comments

This plasmid is compatible with gRNA expression plasmids for budding yeast, such as Addgene Plasmid #43803 ( http://www.addgene.org/43803 ).

For more information on Fujii Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/fujii/