LifeAct-14 (LA-14)
(Plasmid #158750)

• Purpose: Lentiviral plasmid for expression of shorter LifeAct variant (up till 14 amino acids) fused with carboxy terminus eGFP in mammalian cells.
• Depositing Lab: Minhajuddin Sirajuddin
• Publication: Kumari et al EMBO J. 2020 Jun 22:e104006. doi: 10.15252/embj.2019104006.

Backbone

• Vector backbone: pMuLE Dest BlastR
• Backbone manufacturer: Padilla-Rodriguez et al Nat Commun. 2018 Jul 30;9(1):2980. doi: 10.1038/s41467-018-05367-2.
• Backbone size w/o insert (bp): 8467
• Total vector size (bp): 8509
• Modifications to backbone: Inserted LA-14 sequence in the Addgene plasmid # 84383
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: Can grow at 37C also
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: LifeAct-14
• Alt name: LA-14
• Alt name: Shorter variant of actin binding LifeAct (amino acid 1-14)
• Insert Size (bp): 42
• Mutation: Deletion mutation for last three amino acids from LifeAct sequence
• Promoter: CMV immediate early
• Tag / Fusion Protein:
  • eGFP (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: CMV_fwd_primer
• 3′ sequencing primer: MSCV_rev_primer

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Performed site-directed mutagenesis (SDM) to introduced deletion mutations for the terminal 3 amino acids in LifeAct sequence cloned in Addgene plasmid # 84383

Terms and Licenses

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