yeast_DualReporter
(Plasmid #127546)

• Purpose: A dual reporter vector modified from Sharon et al (Nature Biotech, 2012) containing Ura3 and Nat1 selectable markers, a constant background RFP (TEF2::mCherry), and a variable YFP (pTpA+MCS::yEVenus).
• Depositing Lab: Aviv Regev
• Publication: de Boer et al Nat Biotechnol. 2020 Jan;38(1):56-65. doi: 10.1038/s41587-019-0315-8. Epub 2019 Dec 2.

Backbone

• Vector backbone: pPAL1_His3core
• Backbone manufacturer: Sharon et al NBT 2012 (Eran Segal's group)
• Total vector size (bp): 8218
• Modifications to backbone: yEVenus promoter replaced with minimal promoter and multiple cloning site.
• Vector type: Yeast Expression, Synthetic Biology
• Selectable markers: URA3 ; Nourseothricin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: bla
• Alt name: ampicillin resistance
• Alt name: AmpR
• Alt name: beta-lactamase
• Species: bacteria
• Insert Size (bp): 861
• Promoter: bla (E. coli)

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: NA

Gene/Insert 2

• Gene/Insert name: mCherry
• Alt name: RFP
• Species: Synthetic
• Insert Size (bp): 711
• Promoter: TEF2

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: NA

Gene/Insert 3

• Gene/Insert name: URA3
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 804
• Entrez Gene: URA3 (a.k.a. YEL021W)
• Promoter: URA3

Cloning Information for Gene/Insert 3

• Cloning method: Unknown
• 5′ sequencing primer: NA

Gene/Insert 4

• Gene/Insert name: yEVenus
• Alt name: YFP
• Species: Synthetic
• Insert Size (bp): 717
• Promoter: pTpA+MCS

Cloning Information for Gene/Insert 4

• Cloning method: Gibson Cloning
• 5′ sequencing primer: agtgttgttccagtttgg
• 3′ sequencing primer: cggagacagaaaatttgtg

Gene/Insert 5

• Gene/Insert name: Nat1
• Alt name: nourseothricin resistance
• Alt name: nourseothricin acetyltransferase
• Promoter: TEF

Cloning Information for Gene/Insert 5

• Cloning method: Unknown
• 5′ sequencing primer: NA

Resource Information

• Supplemental Documents:
  • ygpra_ptpa_blank.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/224907v3 for bioRxiv preprint.