pAMS366-4xCDRE-GFP-PEST
(Plasmid #138658)

• Purpose: destabilized GFP reporter to monitor calcineurin activity via calcineurin-dependent response element (CDRE) in S. cerevisiae
• Depositing Lab: Sabrina Büttner
• Publication: Diessl et al Microbial Cell, 2020: in press

Backbone

• Vector backbone: pAMS366
• Total vector size (bp): 11281
• Vector type: yeast reporter plasmid
• Selectable markers: URA3

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: 4xCDRE-CYC1(PM)-yEGFP3-PEST
• Species: S. cerevisiae (budding yeast), Synthetic; A. victoria
• Promoter: CYC1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: GTG GGT TTA GAT GAC AAG GGA GAC G and CAC AAA TTT TCT GTC TCC G and GTC TTG TTA CCA GAC AAC C

Resource Information

• A portion of this plasmid was derived from a plasmid made by: 1. Stathopoulos AM, and Cyert MS (1997). Calcineurin acts through the CRZ1/TCN1-encoded transcription factor to regulate gene expression in yeast. Genes Dev. 11(24): 3432–3444. 9407035. and 2. Mateus C, and Avery SV (2000). Destabilized green fluorescent protein for monitoring dynamic changes in yeast gene expression with flow cytometry. Yeast. 16(14): 1313–1323. doi: 10.1002/1097-0061(200010)16:14<1313::AID-YEA626>3.0.CO;2-O.
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

1x CDRE (calcineurin-dependent response element) = caagcgcacagccaccgactggtg

For construction of pAMS366-4xCDRE-GFP-PEST, we used yEGFP3-PEST from
Mateus & Avery 2000 (original yEGFP3 from Cormack et al. 1997),
sequencing showed a 216T>G silent mutation.