pUCC001
(Plasmid #124451)

• Purpose: Multi-species CRISPR/Cas9 coexpression system, optimized for Golden Gate cloning of gRNA targets
• Depositing Lab: John Morrissey
• Publication: Rajkumar et al Front Bioeng Biotechnol. 2019 May 7;7:97. doi: 10.3389/fbioe.2019.00097. eCollection 2019.

Backbone

• Vector backbone: pUDP002
• Backbone manufacturer: TU Delft
• Backbone size w/o insert (bp): 9106
• Total vector size (bp): 10233
• Modifications to backbone: Insert 1 cloned by Gibson assembly between the existing ScTDH3 promoter and ScCYC1 terminator
• Vector type: Yeast Expression
• Selectable markers: Hygromycin

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: HH-BsaI
• Alt name: HH-gRNA-HDV expression cassette
• Species: Synthetic
• Insert Size (bp): 1136
• Mutation: gRNA expression cassette from pUDP002 modified to allow cloning of gRNA targets by Golden Gate assembly
• Promoter: TDH3 promoter from S.cerevisiae (already present in backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TGCTCTCTCTGATTTGGAAAAAGCTGAA
• 3′ sequencing primer: TACACGCGTTTGTACAGAAAAAAAAGAAAAATTTGA

Resource Information

• Supplemental Documents:
  • pUCC001.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The same patent and IP restrictions as in pUDP002 apply here.