pAWH-Rluc-let-7-A114-N40-HhR
(Plasmid #50555)

• Purpose: Expression of luciferase reporter mRNA in S2 cell
• Depositing Lab: Yukihide Tomari
• Publication: Fukaya et al Mol Cell. 2012 Dec 28;48(6):825-36. doi: 10.1016/j.molcel.2012.09.024. Epub 2012 Nov 1.

Backbone

• Vector backbone: pAHW
• Backbone manufacturer: The Drosophila Gateway™ Vector Collection
• Vector type: Insect Expression, Luciferase

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): mv1184
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Luciferase-let7-A114-N40-HhR
• Species: Synthetic
• Promoter: Actin5C

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: RenLuc3'F

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Rluc gene derives from psiCheck2 from Promega.
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

We constructed a Renilla luciferase reporter for let-7 miRNA harboring A114-N40 (a protected poly(A) sequence internalized by a 40 nt unrelated sequence) followed by the hammerhead ribozyme sequence (HhR) that generates the 3' end by self-cleavage.

Unstable when transformed and so any retransformation must be checked by digest or sequencing before use. In strain MV1184, it is stable as a glycerol stock.