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G30, Cdc20-mCherry-AID tagging
(Plasmid #130270)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 130270 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    HO-poly-KanMX4-HO
  • Backbone manufacturer
    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC55758/
  • Backbone size w/o insert (bp) 2531
  • Total vector size (bp) 5973
  • Modifications to backbone
    The HO homologous arms are replaced by Cdc20 homologous arms for genomic integration.
  • Vector type
    Yeast Expression
  • Selectable markers
    NatMX (select with Nourseotrichin)

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
    Cdc20/mCherry-AID(71-114)-tCYC1-NatMX
  • Alt name
    YGL116W
  • Alt name
    PAC5
  • Species
    S. cerevisiae (budding yeast), Synthetic
  • Insert Size (bp)
    2328
  • Entrez Gene
    CDC20 (a.k.a. YGL116W, PAC5)
  • Promoter Native promoter
  • Tags / Fusion Proteins
    • mCherry (C terminal on insert)
    • AID(71-114) (C terminal on insert)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer AGCACTAAGCTGGTGTC
  • 3′ sequencing primer CCATAATGGCGCAGAG
    • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    The AID(71-114) sequence was retrieved from: Morawska, M., and Ulrich, H.D. (2013). An expanded tool kit for the auxin-inducible degron system in budding yeast. Yeast 30, 341–351. mCherry was retrieved from the pBS35 plasmid.

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This plasmid can be used to dynamically and conditionally deplete Cdc20 in yeast.

The work is currently under review.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    G30, Cdc20-mCherry-AID tagging was a gift from Matthias Heinemann (Addgene plasmid # 130270 ; http://n2t.net/addgene:130270 ; RRID:Addgene_130270)

  • For your References section:

    Inference of the High-Level Interaction Topology between the Metabolic and Cell-Cycle Oscillators from Single-Cell Dynamics. Ozsezen S, Papagiannakis A, Chen H, Niebel B, Milias-Argeitis A, Heinemann M. Cell Syst. 2019 Oct 4. pii: S2405-4712(19)30311-4. doi: 10.1016/j.cels.2019.09.003. 10.1016/j.cels.2019.09.003 PubMed 31606371
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