pREC056_EF1a-mCherry::ms2-SV40_CMV-EGFP::pp7-bGH
(Plasmid #262124)

• Purpose: Expresses mCherry::2xms2, EGFP::2xpp7 and tRNA-Pyl (Methanosarcina mazei tRNA-Pyl-CUA) in mammalian cells
• Depositing Lab: Randall Platt
• Depositing Organization: ETH Zurich (Swiss Federal Institute of Technology)
  Located in Switzerland
• Publication: Reinkemeier et al Nat Biotechnol. 2026 Oct 21. doi: 10.1038/s41587-026-03327-6.

Backbone

• Vector backbone: pcDNA3.1/Zeo(+)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5015
• Total vector size (bp): 8758
• Vector type: Mammalian Expression
• Selectable markers: Zeocin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mCherry::ms2
• Species: ms2-bacteriophage, Discosoma sp.
• Insert Size (bp): 786
• Promoter: EF1 alpha
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • 6xHis (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TCAAGCCTCAGACAGTGGTTC

Gene/Insert 2

• Gene/Insert name: EGFP::pp7
• Species: Aequorea victoria
• Insert Size (bp): 819
• Promoter: CMV
• Tags / Fusion Proteins:
  • NLS::FLAG (N terminal on backbone)
  • 6xHis (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CGCAAATGGGCGGTAGGCGTG

Gene/Insert 3

• Gene/Insert name: tRNA-Pyl-CUA
• Species: Methanosarcina mazei
• Insert Size (bp): 69
• Mutation: CUA anticodon
• Promoter: U6

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GAGGGCCTATTTCCCATGATT

Terms and Licenses

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

Depositor Comments

Please consult the Supplemental Document for description of protein sequences for insert and linkers.