pGAN200
(Plasmid #129203)

• Purpose: yTRAP sensor of [PSI+], detects aggregation of Sup35
• Depositing Lab: Ahmad Khalil
• Publication: Newby et al Cell. 2017 Nov 2;171(4):966-979.e18. doi: 10.1016/j.cell.2017.09.041. Epub 2017 Oct 19.

Backbone

• Vector backbone: pGAN147
• Backbone manufacturer: Gregory Newby
• Backbone size w/o insert (bp): 8600
• Total vector size (bp): 9798
• Modifications to backbone: LR reaction with pENTR-NM
• Vector type: Yeast Expression, Synthetic Biology
• Selectable markers: Nourseothricin

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: Sup35NM
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1200
• Mutation: N terminal and middle domains
• Entrez Gene: SUP35 (a.k.a. YDR172W, GST1, PNM2, SAL3, SUF12, SUP2, SUP36)
• Promoter: SUP35

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pGAN200_SUP35NM yTRAP to sense PSI.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • mNeonGreen Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This is the basic yTRAP sensor to sense aggregation of Sup35 or [PSI+]. It should be linearized with NotI-HF enzyme prior to integration into the yeast genome. The sensor integrates into the HO locus, and can be selected for using nourseothricin.