pMM4_14L
(Plasmid #183063)

• Purpose: Plasmid expressing a biosensor for acetic acid and pHluorin: BM3R1-Haa1-mTurquoise2 under RET2p, mCherry under HREsYGP1p and sfpHluorin under TDH3p with flanks for genome integration into HO locus
• Depositing Lab: Yvonne Nygård
• Publication: Mormino et al Microb Cell Fact. 2022 Oct 15;21(1):214. doi: 10.1186/s12934-022-01938-7.

Backbone

• Vector backbone: MoClo-YTK
• Total vector size (bp): 11322
• Vector type: Yeast Expression, Synthetic Biology
• Selectable markers: Zeocin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mCherry
• Species: Synthetic
• Promoter: Synthetic promoter
• Tag / Fusion Protein:
  • mCherry

Cloning Information for Gene/Insert 1

• Cloning method: Gateway Cloning
• 5′ sequencing primer: Golden Gate Assembly

Gene/Insert 2

• Gene/Insert name: sfpHluorin
• Species: Synthetic
• Promoter: TDH3

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: Golden Gate Assembly

Gene/Insert 3

• Gene/Insert name: BM3R1-HAA1-mTurquoise2
• Species: Synthetic
• Promoter: RET2
• Tag / Fusion Protein:
  • mTurquoise2

Cloning Information for Gene/Insert 3

• Cloning method: Gateway Cloning
• 5′ sequencing primer: Golden Gate Assembly

Resource Information

• Supplemental Documents:
  • MM4_14L - pRET2 - integrated - Zeocin yeast resistance - strong sf pHluorin.gbk

Terms and Licenses

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

Depositor Comments

Plasmid parts were synthesized or taken from the Yeast MoClo kit. A portion of this plasmid was derived from LT1_34_sfpHluorin that was partly made by Digital CDS sequence of sfpHluorin obtained from Addgene plasmid #115697.