pGHUC w-ERBIN (2-hybrid prey plasmid)
(Plasmid #128163)

• Purpose: UV5 promoter drives expression of omega-erbin fusion. Pair with pB1H2 UV5 Zif268-cons as a positive control (turns on HIS3/GFP).
• Depositing Lab: Marcus Noyes
• Publication: Ichikawa et al ACS Synth Biol. 2019 May 17;8(5):918-928. doi: 10.1021/acssynbio.8b00499. Epub 2019 Apr 18.

Backbone

• Vector backbone: THE pGHUC
• Backbone size w/o insert (bp): 7715
• Total vector size (bp): 8042
• Modifications to backbone: Original backbone as published in: Oakes et al. Multi-reporter selection for the design of active and more specific zinc-finger nucleases for genome editing. Nature Communications. 2016. doi: 10.1038/ncomms10194. Modifications: Nonessential F1 ori replaced with UV5 promoter driving omega-ERBIN fusion. Sequence between EcoRI and AgeI replaced. Two copies of the preferred zif268 binding site were inserted consecutively, but in opposite orientations, 21bp upstream of the -35 boxes of relevant reporter promoters.
• Vector type: Bacterial Expression, Synthetic Biology

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: ERBIN PDZ domain
• Species: Synthetic
• Insert Size (bp): 327
• Entrez Gene: ERBIN (a.k.a. ERBB2IP, HEL-S-78, LAP2)
• Promoter: UV5
• Tags / Fusion Proteins:
  • FLAG (N terminal on backbone)
  • Omega subunit of RNA polymerase (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: CAAGAGCAGGAAGCCGCTG

Resource Information

• Supplemental Documents:
  • pGHUC w-ERBIN (2-hybrid prey plasmid).gbk

Terms and Licenses

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