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pKPY197
(Plasmid #62599)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 62599 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pPD95.75
  • Backbone manufacturer
    Andrew Fire (Addgene Plasmid #1494)
  • Backbone size w/o insert (bp) 4494
  • Total vector size (bp) 6351
  • Modifications to backbone
    The gene encoding the mutant alpha subunit of C. elegans PheRS was isolated by PCR amplification. The intercistronic region of the C. elegans operon CEOP5428 as described by Lee et al. (Lee, L.W., Lo, H.W. and Lo, S.J. Gene 455, 16–21 (2010)) was isolated by genomic DNA extraction from wild-type (N2) C. elegans and PCR amplification. The purified fragments were ligated into pPD95.75 (SalI, KpnI) to generate pKPY197. Note: the first methionine in the alpha subunit of C. elegans PheRS, Isoform A, Exon 3 is designated as residue 1 (“Met1”).
  • Vector type
    Worm Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    XL1 Blue
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Thr412Gly-CePheRS Alpha Subunit
  • Alt name
    Alpha Subunit of C. elegans Phenylalanyl-tRNA Synthetase, Isoform A (Thr412Gly)
  • Species
    C. elegans (nematode)
  • Insert Size (bp)
    1957
  • Mutation
    Thr412Gly
  • Tag / Fusion Protein
    • HIS (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SalI (not destroyed)
  • 3′ cloning site KpnI (not destroyed)
  • 5′ sequencing primer M13 Reverse
  • 3′ sequencing primer GFP-R
  • (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:

    pKPY197 was a gift from David Tirrell (Addgene plasmid # 62599)
  • 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