pKPY514
(Plasmid #62598)

• Purpose: Encodes Thr251Gly-EcPheRS Under IPTG-Inducible (PT5) Control
• Depositing Lab: David Tirrell
• Publication: Yuet et al Proc Natl Acad Sci U S A. 2015 Feb 17. pii: 201421567.

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, 50 μg/mL
• 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

Resource Information

• Supplemental Documents:
  • KY33[pKPY514]
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Please note that this plasmid may require a unique bacterial strain, so make sure to confirm that you can also obtain the appropriate growth strain. Please contact us at or contact our distributors if you have any questions.