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pZE21/UBP1/ClpS_V65I
(Plasmid #98567)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 98567 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pZE21
  • Backbone manufacturer
    Expressys
  • Backbone size w/o insert (bp) 2200
  • Total vector size (bp) 4696
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Note that this plasmid, unlike the related pZE21/UBP1/ClpS plasmid, does contain the Tet repressor cloned into the pZE21 backbone downstream of ClpS and under constitutive expression. The plasmid map provided may not reflect this.
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    Truncated ubiquitin cleavase, codon optimized
  • Alt name
    UBP1
  • Species
    S. cerevisiae (budding yeast)
  • Insert Size (bp)
    2151
  • Mutation
    Removed the first 98 amino acids that encode transmembrane domains to improve yield as was done in literature (Wojtowicz et al, Microb Cell Fact, 2005, DOI: 10.1186/1475-2859-4-17)

Gene/Insert 2

  • Gene/Insert name
    Mutated ATP-dependent Clp protease adapter protein
  • Alt name
    ClpS_V65I
  • Species
    Escherichia coli
  • Insert Size (bp)
    318
  • Mutation
    Contains the V65I mutation that increases discrimination between N-terminal Tyr/Phe compared to N-terminal non-standard amino acids. In a synthetic operon downstream of UBP1

Resource Information

Depositor Comments

Addgene NGS results found H571Y in the UBP1 translation compared to NP_010161.1. The depositing lab confirms that this mutation should not affect protein function

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:

    pZE21/UBP1/ClpS_V65I was a gift from George Church (Addgene plasmid # 98567 ; http://n2t.net/addgene:98567 ; RRID:Addgene_98567)
  • For your References section:

    Engineering posttranslational proofreading to discriminate nonstandard amino acids. Kunjapur AM, Stork DA, Kuru E, Vargas-Rodriguez O, Landon M, Soll D, Church GM. Proc Natl Acad Sci U S A. 2018 Jan 16;115(3):619-624. doi: 10.1073/pnas.1715137115. Epub 2018 Jan 4. 10.1073/pnas.1715137115 PubMed 29301968