pSP148
(Plasmid #247110)

• Purpose: Expresses estrogen-binding domain-GFP-nanobody-Rnf4 fusion protein in Aspergillus fumigatus
• Depositing Lab: Scott Moye-Rowley
• Publication: Paul et al G3 (Bethesda). 2026 Feb 4;16(2):jkaf295. doi: 10.1093/g3journal/jkaf295.

Backbone

• Vector backbone: pUC57
• Vector type: Bacterial Expression, Affinity Reagent/ Antibody ; Maintenance in bacteria

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: Human estrogen-binding domain from the estrogen receptor
• Species: H. sapiens (human)
• Insert Size (bp): 942
• Entrez Gene: ESR1 (a.k.a. ER, ESR, ESRA, ESTRR, Era, NR3A1)
• Tag / Fusion Protein:
  • 4X HA (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gene Synthesis
• 5′ sequencing primer: Nanopore whole plasmid sequencing used to verify

Gene/Insert 2

• Gene/Insert name: GFP Nanobody
• Species: Camellid
• Insert Size (bp): 354

Cloning Information for Gene/Insert 2

• Cloning method: Gene Synthesis
• 5′ sequencing primer: Nanopore whole plasmid sequencing used to verify

Gene/Insert 3

• Gene/Insert name: Rat Rnf4
• Species: R. norvegicus (rat)
• Insert Size (bp): 360
• Entrez Gene: Rnf4

Cloning Information for Gene/Insert 3

• Cloning method: Gene Synthesis
• 5′ sequencing primer: Nanopore whole plasmid sequencing used to verify

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The gpdA-driven reverse tet transactivator protein and tet-on-minimal promoter were obtained from Dr. Fabio Gsaller and the construction of these two features is described here: https://pubmed.ncbi.nlm.nih.gov/36350143/ .The 4X HA-GFPNb-rnf4 fusion gene was synthesized with A. fumigatus codon bias by Genscript. The GFP nanobody sequence-Rnf4 fusion was taken directly from: https://pmc.ncbi.nlm.nih.gov/articles/PMC7332993/ .