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LHP809 - Ub-K63A-CYC1
(Plasmid #32166)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 32166 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    YEplac195
  • Modifications to backbone
    Contains CUP1 promoter, synthetic yeast ubiquitin, and CYC1 terminator sequence - all cloned in at same time as described in comments. K63A mutation was introduced by site-directed mutagenesis of LHP585 (Addgene 32168).
  • Vector type
    Yeast Expression
  • Selectable markers
    URA3

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
    synthetic yeast Ubiquitin
  • Species
    S. cerevisiae (budding yeast)
  • Mutation
    K63A
  • Entrez Gene
    UBA1 (a.k.a. YKL210W)
  • Entrez Gene
    UBA2 (a.k.a. YDR390C, UAL1)
  • Promoter CUP1

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site none (unknown if destroyed)
  • 3′ cloning site none (unknown if destroyed)
  • 5′ sequencing primer CUP1 forward
  • 3′ sequencing primer LacZ-R
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

A ~1040 bp BamHI / HindIII fragment containing the CUP1 promoter, the synthetic yeast Ub gene and a CYC1 terminator sequence was ligated into the BamHI and HindIII sites of YEplac195. The resulting plasmid is approximately 6.281kb).

K63A mutation was introduced by site-directed mutagenesis of LHP585 (Addgene 32168).

2 µm S. cerevisiae replication.

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:

    LHP809 - Ub-K63A-CYC1 was a gift from Linda Hicke (Addgene plasmid # 32166 ; http://n2t.net/addgene:32166 ; RRID:Addgene_32166)
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