pMuLE_ENTR_TOP-NLuc1.1_L5-L4
(Plasmid #113867)

• Purpose: Single pathway reporter; wnt promoter-driven expression of intracellular NanoLuc; MultiSite Gateway entry vector.
• Depositing Lab: Manfred Ogris
• Publication: Maier et al PLoS One. 2019 Dec 20;14(12):e0226570. doi: 10.1371/journal.pone.0226570. eCollection 2019.

Backbone

• Vector backbone: pMuLE ENTR MCS L5-L4
• Backbone size w/o insert (bp): 2665
• Total vector size (bp): 3693
• Vector type: Mammalian Expression ; MuLE Gateway Entry Vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: TOP (wnt dependent promoter)
• Alt name: M50 Super 8x TOPFlash, promoter only
• Insert Size (bp): 214

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: M13

Gene/Insert 2

• Gene/Insert name: NLuc
• Alt name: NL1.1
• Alt name: NanoLuc, intracellular
• Insert Size (bp): 822

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: M13

Resource Information

• Supplemental Documents:
  • pMuLE_ENTR_TOP-NLuc1.1_L5-L4.gb
  • Plasmid History
• A portion of this plasmid was derived from a plasmid made by: M50 Super 8x TOPFlash (Addgene plasmid #12456) was a gift from Randall Moon. Promoterless pNL1.1 (#N1001) was obtained from Promega. Backbone is included in the MuLE kit, which was a gift from Ian Frew (Addgene kit #1000000060).

Terms and Licenses

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

Depositor Comments

The PTCH1 promoter contains a C3831G mutation compared to the NCBI reference that is not of concern for plasmid function.