pUC19mut_circIRES10_FLuc_3HA
(Plasmid #249692)

• Purpose: This plasmid is designed to evaluate the translation efficiency of IRES10, an internal ribosome entry site derived from the Hibiscus chlorotic virus (HCRSV).
• 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• Publication: Schlemmer Lab Plasmids 2025 (unpublished)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 Hibiscus chlorotic ringspot virus
• Alt name: IRES10
• Species: Hibiscus chlorotic ringspot virus
• Insert Size (bp): 101
• 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 IRES10, an internal ribosome entry site derived from the Hibiscus chlorotic virus (HCRSV). This IRES is located upstream of the coat protein (CP) gene and features a relatively simple secondary structure. Studies have shown that each half of the IRES can independently initiate translation, and a possible interaction with the 3' UTR has been demonstrated to enhance translation in bicistronic constructs (Koh, Wong, and Liu 2003). 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.