pEVOL-pBpF
(Plasmid #31190)

• Purpose: tRNA synthetase/tRNA pair for the in vivo incorporation of a photocrosslinker, p-benzoyl-l-phenylalanine into proteins in E coli in response to the amber codon, TAG.
• Depositing Lab: Peter Schultz
• Publication: Chin et al Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11020-4. Epub 2002 Aug 1.

Backbone

• Vector backbone: p15A
• Backbone size w/o insert (bp): 5000
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Any strain for growth, 37 oC, LB/2XYT media
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: M.j. p-benzoylphenylalanine RS (2 copies +tRNA)
• Alt name: pEVOL-pBpF
• Species: M.jannaschii
• Insert Size (bp): 1000
• Mutation: Y32G, E107P, D158T, I159S

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: ATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCCGTTTTTT

Resource Information

• Addgene Notes:
  • Additional Addgene Diagnostic digest
• Articles Citing this Plasmid:
  • 45 References

Terms and Licenses

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