pEVOL_PylRS(AF)-tRNA
(Plasmid #223511)

• Purpose: PylRS (AF)/tRNA Pyl orthogonal pair for genetic code expansion that allows site-specific incorporation of noncanonical amino-acids into a POI using the Amber stop codon
• Depositing Lab: Edward Lemke
• Publication: Milles et al J Am Chem Soc. 2012 Mar 21;134(11):5187-95. doi: 10.1021/ja210587q. Epub 2012 Mar 5.

Backbone

• Vector backbone: pEVOL
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Pyrrolysyl-tRNA Synthetase (AF) (2 copies)
• Species: Methanosarcina mazei
• Insert Size (bp): 2730
• Mutation: Y306A; Y384F
• Entrez Gene: pylS (a.k.a. MM_RS07465, MM_1445)
• Promoter: araBAD, rrnB, and glnS
• Tag / Fusion Protein:
  • 6xHis (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: tRNA Pyl
• Species: Methanosarcina mazei
• Insert Size (bp): 72
• Promoter: proK

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Supplemental Documents:
  • pEVOL_PylRS(AF)-tRNA.gb

Terms and Licenses

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