pKPY197
(Plasmid #62599)

• Purpose: Encodes Thr412Gly-CePheRS Alpha Subunit
• Depositing Lab: David Tirrell
• Publication: Yuet et al Proc Natl Acad Sci U S A. 2015 Feb 17. pii: 201421567.

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, 100 μg/mL
• 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

Resource Information

• Supplemental Documents:
  • XL1-Blue[pKPY197]
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry