pUC19mut_circIRES7_FLuc_3HA
(Plasmid #249690)

• Purpose: This plasmid is designed to evaluate the translation efficiency of IRES7, a internal ribosome entry site from the Crucifer infecting tobamovirus (crTMV).
• Depositing Lab: Timo Schlemmer
• Depositing Organization: Universitaet Regensburg
  Located in Germany
• Publication: Schlemmer Lab Plasmids 2025 (unpublished)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 Crucifer infecting tobamovirus
• Alt name: IRES7
• Species: Crucifer tobamovirus
• Insert Size (bp): 75
• 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 IRES7, a internal ribosome entry site from the Crucifer infecting tobamovirus (crTMV). Located upstream of the coat protein (CP) ORF, this A-rich IRES features a proposed secondary structure of two stem-loops separated by a purine-rich tract with repetitive elements (Ivanov et al., 1997). This tract is credited with giving the IRES its significantly high translational activity (Dorokhov et al., 2002). 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.