LifeAct-mNeonGreen -IRES- Talin head-SpyTag003
(Plasmid #210024)

• Purpose: Expression of LifeAct-mNeonGreen and Talin head(1-433)-SpyTag003 in mammalian cells
• Depositing Lab: Vesa Hytönen
• Publication: Rahikainen et al J Am Chem Soc. 2023 Nov 15;145(45):24459-24465. doi: 10.1021/jacs.3c07827. Epub 2023 Oct 31.

Backbone

• Vector backbone: Modified pcDNA3 (1xU6-PylT, 1xH1-PylT)
• Backbone manufacturer: Described in PMID: 33069552
• Backbone size w/o insert (bp): 5959
• Total vector size (bp): 8383
• Modifications to backbone: One copy of Desulfitobacterium hafniense pyrrolysyl-tRNACUA (PylT) with G8U under the control of a U6 promoter (1× U6-PylT) and one copy of Methanosarcina barkeri PylT with U25C under the control of a H1 promoter (1× H1-PylT)
• 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: LifeAct-mNeonGreen co-expressed with Talin head(1-433)-SpyTag003
• Species: M. musculus (mouse), Synthetic
• Insert Size (bp): 2751
• Mutation: Encephalomyocarditis virus internal ribosome entry site (IRES) with 6A bifurcation loop to improve translation efficiency of 3' ORF
• Promoter: CMV

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: acgcaaatgggcggtagg
• 3′ sequencing primer: atttaggtgacactatag

Resource Information

• Supplemental Documents:
  • LifeAct-mNeonGreen_IRES_TalHead-SpyTag003.gbk

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • mNeonGreen Label License
• Industry Terms:
  • Not Available to Industry