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pKPY514
(Plasmid #62598)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 62598 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pQE-80L-Kan
  • Backbone manufacturer
    Qiagen
  • Backbone size w/o insert (bp) 4541
  • Total vector size (bp) 7897
  • Modifications to backbone
    The genes encoding wild-type E. coli PheRS were isolated by genomic DNA extraction from E. coli DH10B and PCR amplification. The purified fragments were ligated into pQE-80L-Kan (SphI, HindIII). The resulting plasmid was subjected to site-directed mutagenesis to generate the Thr251Gly mutation.
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    KY33
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Thr251Gly-EcPheRS
  • Alt name
    E. coli Phenylalanyl-tRNA Synthetase (Thr251Gly)
  • Species
    E. coli
  • Insert Size (bp)
    3386
  • Mutation
    Thr251Gly
  • Promoter T5
  • Tag / Fusion Protein
    • 6xHis (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SphI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer pQE Promoter
  • 3′ sequencing primer pQE Reverse
  • (Common Sequencing Primers)

Resource Information

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:

    pKPY514 was a gift from David Tirrell (Addgene plasmid # 62598)
  • For your References section:

    Cell-specific proteomic analysis in Caenorhabditis elegans. Yuet KP, Doma MK, Ngo JT, Sweredoski MJ, Graham RL, Moradian A, Hess S, Schuman EM, Sternberg PW, Tirrell DA. Proc Natl Acad Sci U S A. 2015 Feb 17. pii: 201421567. 10.1073/pnas.1421567112 PubMed 25691744