pRS405 SEC66-VAC8-RFP
(Plasmid #166843)

• Purpose: Expresses ER localized Vac8-RFP in yeasts. Made replacing the first 21 base pairs of the VAC8 ORF with the first 162 base pairs from the SEC66 ORF, which encode the Sec66 transmembrane domain.
• Depositing Lab: Daniel Klionsky
• Publication: Gatica et al Autophagy. 2020 Jun 8. doi: 10.1080/15548627.2020.1776474.

Backbone

• Vector backbone: pRS405
• Total vector size (bp): 8453
• Vector type: Yeast Expression
• Selectable markers: LEU2

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: VAC8
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 2544
• Mutation: SEC66 transmembrane domain
• Entrez Gene: VAC8 (a.k.a. YEL013W, YEB3)
• Entrez Gene: SEC66 (a.k.a. YBR171W, HSS1, KAR7, SEC71)
• Promoter: VAC8 promoter
• Tags / Fusion Proteins:
  • RFP (C terminal on insert)
  • SEC66 transmembrane domain (N terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: M13F
• 3′ sequencing primer: M13R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Linearize with AflII