pUC57-Rluc-let-7-A114-N40
(Plasmid #50563)

• Purpose: Template DNA for in vitro transcription using T7 polymerase
• Depositing Lab: Yukihide Tomari
• Publication: Fukaya et al EMBO J. 2011 Nov 25;30(24):4998-5009. doi: 10.1038/emboj.2011.426.

Backbone

• Vector backbone: pUC57
• Backbone manufacturer: GeneScript
• Backbone size w/o insert (bp): 2710
• Total vector size (bp): 4016
• Vector type: cloning vector

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
• Species: Renilla, unknown
• Promoter: lac

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: M13R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Rluc gene derives from psiCheck2 from Promega.

Terms and Licenses

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

Depositor Comments

Plasmid contains 8 copies of the let-7 target site, a defined A114 sequence followed by an EcoT22I site and a 40-bp arbitrary sequence all downstream of Renilla lucferase.

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. Please note that the Rluc coding sequence contains an EcoT22I site. To prepare the T7 transcription template for A114 RNA (without N40), the plasmid should be first linearized by EcoRI upstream of the T7 promoter sequence, followed by a partial digestion by EcoT22I. The DNA fragment terminating immediately downstream of the A114 sequence should then be gel purified. For A114-N40 RNA, the plasmid should be digested by HindIII downstream of the N40 sequence.