pDR111_mannose_Stat1Klf6
(Plasmid #188400)

• Purpose: Expresses mouse transcription factors Stat-1 and Klf6 in an operon from D-mannose promoter for Bacillus subtilis. The secretion peptide PhoD is fused to both factors.
• Depositing Lab: Christopher Contag
• Publication: Madsen et al Commun Biol. 2022 Aug 30;5(1):888. doi: 10.1038/s42003-022-03851-6.

Backbone

• Vector backbone: pDR111
• Backbone size w/o insert (bp): 8858
• Total vector size (bp): 12312
• Modifications to backbone: Addition of insert and fusion components to NheI site. Phyper-spank replaced by D-mannose promoter and lacI replaced by mannose regulator
• Vector type: Bacterial Expression
• Selectable markers: Spectinomycin for Bacillus subtilis

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Stat-1
• Species: M. musculus (mouse); Escherichia coli
• Insert Size (bp): 2250
• Entrez Gene: Stat1 (a.k.a. 2010005J02Rik)
• Promoter: D-mannose
• Tag / Fusion Protein:
  • PhoD secretion peptide (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: EagI (not destroyed)
• 5′ sequencing primer: cggctagcTTACAGATGCCTT
• 3′ sequencing primer: TACCTCCTTgtcgactAAGCTT

Gene/Insert 2

• Gene/Insert name: Klf6
• Species: M. musculus (mouse)
• Insert Size (bp): 850
• Entrez Gene: Klf6 (a.k.a. AI448727, BCD1, C86813, CPBP, Copeb, FM2, FM6, Ierepo1, Ierepo3, R75280, Zf9)
• Promoter: D-mannose
• Tag / Fusion Protein:
  • PhoD secretion peptide

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: EagI (not destroyed)
• 3′ cloning site: NheI (not destroyed)
• 5′ sequencing primer: cggctagcTTACAGATGCCTT
• 3′ sequencing primer: TACCTCCTTgtcgactAAGCTT

Resource Information

• Supplemental Documents:
  • pdr111_mannose_m1operon_restrict_v2 (1).gb

Terms and Licenses

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