pNEU-hMbPylRS-4xU6M15
(Plasmid #105830)

• Purpose: Plasmid encoding for the Pyrrolysine translational pair that enables efficent incoporation of Pyl-like click amino acids in response to the amber codon into proteins in mammalian cells
• Depositing Lab: Irene Coin
• Publication: Serfling et al Nucleic Acids Res. 2017 Nov 21. pii: 4641905. doi: 10.1093/nar/gkx1156.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Humanized MbPylRS
• Species: Synthetic; M. barkeri
• Mutation: (Y271A, Y349F)MbPylRS
• Promoter: CMV

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: CMV-F

Gene/Insert 2

• Gene/Insert name: tRNAM15
• Species: Synthetic
• Promoter: U6

Cloning Information for Gene/Insert 2

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

Resource Information

• Supplemental Documents:
  • pNEU-hMbPylRS(AF)-4xU6M15.gb
• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: The codon-optimized gene of MbPylRSF was custom synthesized by GeneArt (ThermoFisher Scientific).
• Articles Citing this Plasmid:
  • 8 References

Terms and Licenses

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

Depositor Comments

The plasmid contains a novel tRNA that boosts the efficiency of the Pyrrolysyl system. Note: Addgene found that there are only 3x repeats of U6M15 instead of 4x as noted in the publication.