pUC19mut_circIRES4_FLuc_3HA
(Plasmid #249687)

• Purpose: This plasmid is designed to evaluate the translation efficiency of IRES4, an internal ribosome entry site derived from the Pelargonium flower break virus.
• 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• Publication: Schlemmer Lab Plasmids 2025 (unpublished)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 Pelargonium flower break virus
• Alt name: IRES4
• Species: Pelargonium flower break virus
• Insert Size (bp): 77
• 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 IRES4, an internal ribosome entry site derived from the Pelargonium flower break virus. The IRES is located within an fragment upstream of the open reading frame for the p37 protein and features a relatively simple secondary structure, proposed to consist of two hairpin structures and one stem-loop (Fernandez-Miragall and Hernandez, 2011). 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.