pUC19mut_circIRES9_FLuc_3HA
(Plasmid #249691)

• Purpose: This plasmid is designed to evaluate the translation efficiency of IRES9, an internal ribosome entry site derived from the AU-rich 5' UTR of the Turnip mosaic potyvirus (TuMV).
• Depositing Lab: Timo Schlemmer
• Publication: Schlemmer Lab Plasmids 2025 (unpublished)

Backbone

• Vector backbone: pUC19mut
• Backbone manufacturer: Selfmade
• Backbone size w/o insert (bp): 4642
• Vector type: Bacterial Expression, Synthetic Biology

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

• Gene/Insert name: IRES of Turnip mosaic potyvirus
• Alt name: IRES9
• Species: Turnip mosaic virus/ Potyvirus rapae
• Insert Size (bp): 134
• Promoter: T7 Promoter
• Tag / Fusion Protein:
  • 3xHA (C terminal on insert)

Cloning Information

• Cloning method: Golden Gate

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid is designed to evaluate the translation efficiency of IRES9, an internal ribosome entry site derived from the AU-rich 5' UTR of the Turnip mosaic potyvirus (TuMV). This IRES initiates the translation of the viral polyprotein, with studies suggesting a mechanism where ribosomes bind to a specific site before scanning for the start codon. Notably, its function appears to be independent of the primary RNA sequence, as its activity is not negatively affected when replaced with its reverse complimentary counterpart (Basso et al., 1994). The firefly luciferase (Fluc) reporter gene is positioned downstream of the IRES to enable quantitative analysis of translation efficiency through luciferase assays. A C-terminal 3xHA tag allows for the detection of Fluc expression via western blotting. This construct is primarily intended for comparative luciferase assays to assess the relative efficiency of different IRES elements in driving translation. Firefly luciferase downstream of IRES to asses translation efficiency.