pUC19mut_circIRES1_FLuc_3HA
(Plasmid #249684)

• Purpose: This plasmid is designed to evaluate the translation efficiency of IRES1, an internal ribosome entry site derived from Blackcurrant reversion virus (BRV).
• 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• Publication: Schlemmer Lab Plasmids 2025 (unpublished)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 Blackcurrant reversion virus
• Alt name: IRES1
• Species: Blackcurrant reversion virus/ Nepovirus ribis
• Insert Size (bp): 161
• 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 IRES1, an internal ribosome entry site derived from Blackcurrant reversion virus (BRV). BRV is a Comoviridae virus with a bipartite positive-sense RNA genome; the 161-nt 5_ UTR of RNA2 functions as a cap-independent IRES and contains regions complementary to plant 18S rRNA, consistent with direct ribosome recruitment (Karetnikov & Lehto, 2007). The firefly luciferase (Fluc) reporter is positioned downstream of IRES1 to provide a quantitative readout in luciferase assays, and a C_terminal 3_HA tag enables detection of Fluc by western blot. The construct is intended for comparative luciferase assays benchmarking the relative efficiency of IRES1 against alternate IRES elements, sequence variants, or conditions that modulate cap-independent translation. Firefly luciferase downstream of IRES to asses translation efficiency.