pRS42H_PTEF1_dCas9_TCYC1
(Plasmid #177155)

• Purpose: Expresses dCas9 in yeast cells
• Depositing Lab: Gautam Dantas
• Publication: Kwak et al ACS Synth Biol. 2022 Jan 21;11(1):508-514. doi: 10.1021/acssynbio.1c00384. Epub 2021 Dec 23.

Backbone

• Vector backbone: pRS42H
• Backbone manufacturer: EUROSCARF
• Backbone size w/o insert (bp): 6798
• Total vector size (bp): 10938
• Modifications to backbone: Added TEF1 promoter and CYC1 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: Unknown

Gene/Insert

• Gene/Insert name: Nuclease-null Cas9
• Alt name: dCas9
• Species: Synthetic
• Insert Size (bp): 4140
• Promoter: TEF1 promoter
• Tag / Fusion Protein:
  • SV40 NLS (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: T3
• 3′ sequencing primer: T7

Resource Information

• Supplemental Documents:
  • prs42h-ptef1-dcas9-tcyc1.gb
• A portion of this plasmid was derived from a plasmid made by: Addgene #63801

Terms and Licenses

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

Depositor Comments

Please note there is a single mutation L326P in HygR. Depositor confirms this mutation does not affect the resistance to hygromycin B.