pSupAR-MbPylRS(DiZPK)
(Plasmid #91705)

• Purpose: express Methanosarcina barkeri pyrrolysyl-tRNA synthetase/tRNA pair in E. coli to incorporate photocrosslinkers DiZPK, DiZSeK, or DiZHSeC into an in-frame amber codon site on a protein of interest
• Depositing Lab: P. Chen
• Publication: Zhang et al Nat Chem Biol. 2011 Sep 4;7(10):671-7. doi: 10.1038/nchembio.644.

Backbone

• Vector backbone: pSupAR
• 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: pylS
• Species: Methanosarcina barkeri
• Insert Size (bp): 1260
• Mutation: T810G, C813T, C820G, T821C, T937A, A1046T, G1257A

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Gene/Insert 2

• Gene/Insert name: pylT
• Species: Methanosarcina barkeri

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • Annotated SnapGene file

Terms and Licenses

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

Depositor Comments

Plasmid for encoding the Methanosarcina barkeri pyrrolysyl-tRNA synthetase (MbPylRS)/tRNA pair in E. coli which can incorporate the photocrosslinkers DiZPK, DiZSeK, or DiZHSeC into a in-frame amber codon site on a protein of interest.The aminoacyl-tRNA synthetase is engineered to recognize the UAA but not the natural native amino acids, and charge its cognate tRNA with the corresponding UAA. Then, the charged tRNA will insert the UAA at an in-frame amber codon position on a target gene.