MmXYRS-4xM15
(Plasmid #155343)

• Purpose: Plasmid encoding the translational pair that enables efficient incorporation of multiple phenylalanine-based amino acids in response to the amber codon into proteins in mammmalian cells.
• Depositing Lab: Irene Coin
• Publication: Bottke et al EMBO Rep. 2020 Sep 14:e50437. doi: 10.15252/embr.202050437.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen, major modifications
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Humanized MmXYRS/tRNAM15
• Species: Synthetic; M. mazei
• Promoter: CMV/U6

Cloning Information

• 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:
  • pNEU_CMV-hMmXYRS-4xU6-M15.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The plasmid contains the MmXYRS, which incorporates ncAAs for chemical crosslinking, and a novel tRNA that boosts the efficiency of the Pyrrolysyl system.

References: Böttke et al, EMBO reports, 2020 and Serfling, R. et al. Designer tRNAs for efficient incorporation of non-canonical amino acids by the pyrrolysine system in mammalian cells. Nucleic Acids Res. 10.1093/nar/gkx1156 (2017).