pLacO-pEF1α-RFP
(Plasmid #179507)

• Purpose: Expresses monomeric RFP in mammalian cells under a EF1α promoter
• Depositing Lab: Yasushi Hiraoka
• Publication: Haraguchi et al Commun Biol. 2022 Jan 20;5(1):78. doi: 10.1038/s42003-022-03021-8.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4731
• Total vector size (bp): 15000
• Modifications to backbone: replace CMV promoter for EF1α
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: Recommend plate culture because liquid culture causes the reduction of the lacO repeat number.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: monomeric RFP
• Alt name: mRFP
• Species: Discosoma sp. mushroom
• Insert Size (bp): 675
• GenBank ID:
• Promoter: EF1α

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: BglII (not destroyed)
• 5′ sequencing primer: -
• 3′ sequencing primer: -

Resource Information

• Supplemental Documents:
  • EF1a-mRFP-pEGFPC1.gb
• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please note, the exact number of Lac operators could not be determined via sequencing. Please refer to the diagnostic digest with AgeI-HF and SalI-HF confirming a ~7kb band representing approximately 200 copies of the lac operator within the plasmid.