pCYC1pr-GNSI (LEU)
(Plasmid #111450)

• Purpose: GNSI is a synthetic, genetically-encoded reporter that allows rapid plate-based assessment of AP-3 functional deficiency, using either colorimetric or growth phenotype readouts.
• Depositing Lab: Alexey Merz
• Publication: Plemel et al Traffic. 2020 Nov 23. doi: 10.1111/tra.12772.

Backbone

• Vector backbone: pRS415
• Backbone size w/o insert (bp): 6021
• Total vector size (bp): 8692
• 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: Low Copy

Gene/Insert 1

• Gene/Insert name: CYC1 Promoter
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 290
• Entrez Gene: CYC1 (a.k.a. YJR048W)

Gene/Insert 2

• Gene/Insert name: Nyv1 Cytosolic Domain (aa 6-230)
• Alt name: GNS cassette
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 675
• Mutation: M109I (Naturally occurring SNP)
• Tag / Fusion Protein:
  • mGFP5 (N terminal on insert)

Gene/Insert 3

• Gene/Insert name: Snc1 Transmembrane Domain (TMD)
• Alt name: GNS cassette
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 60

Gene/Insert 4

• Gene/Insert name: SUC2 CDS
• Alt name: GSI cassette
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1539
• Mutation: The first 21 residues are deleted, removing the localization signal sequence

Gene/Insert 5

• Gene/Insert name: SUC2 terminator
• Alt name: GSI cassette
• Insert Size (bp): 459

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pGNS416, encoding GFP-Nyv1-Snc1TMD (GNS cassette) was a gift from F. Reggiori {Reggiori et al., 2000 PMID 11071903}. pHB4, encoding GSI (in earlier work called GSS) was a gift from E. Conibear {Burston et al., 2009, Addgene #53462}.

Terms and Licenses

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

Depositor Comments

bioRxiv pre-print 314286: Plate-based assay of AP-3 traffic in budding yeast
Please visit https://www.biorxiv.org/content/10.1101/314286v2