Botman-Teusink Yeast FP Collection
This comprehensive collection of yeast codon-optimized fluorescent proteins (FPs) in a variety of plasmids allows for constitutive overexpression of FPs and tagging of genes of interest with FPs in yeast. Characterization of some of the FPs in this collection can be found in Botman et al., 2019.
FP Tagging
Yeast tagging vectors to create fusions of proteins of interest with yeast codon-optimized fluorescent proteins. These vectors are based on the pFA6a-link plasmid (Sheff et al., 2004; Lee et al., 2013), and are available with URA3, SpHIS5 and KANMX markers.
The FP together with the protothropic cassette can be amplified with the following primers:
- FW 5’–GGTGACGGTGCTGGTTTA–’3
- RV 5’–TCGATGAATTCGAGCTCG–’3
| ID | Plasmid | Selectable Marker | Tags | Publication |
|---|
FP Overexpression
Plasmids for constitutive overexpression of codon-optimized FPs in yeast. Vectors are based on the pDRF1 plasmid (Loqué et al., 2007) containing the PMA1 promoter and the URA3 marker, the p415 plasmid (Loqué et al., 2007) containing the PMA1 promoter and the LEU2 marker, or the pCEV-G2-Km plasmid (Vickers et al., 2013) containing the TEF1 or PGK1 promoters and the KanMX marker.
| ID | Plasmid | Gene/Insert | Selectable Marker | Publication |
|---|
Additional Resources
Kits
- MoClo-YTK
- Yeast Gateway Kit
- Multiplex Yeast Toolkit (MYT)
- Markerless Yeast Localization and Overexpression (MyLO) CRISPR-Cas9 Toolkit
Other Plasmids
External Resources
References
- Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B. In vivo characterisation of fluorescent proteins in budding yeast. Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z. PubMed 30783202 (Link opens in a new window)
- Lee S, Lim WA, Thorn KS. Improved blue, green, and red fluorescent protein tagging vectors for S. cerevisiae. PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902. PubMed 23844123 (Link opens in a new window)
- Loqué D, Lalonde S, Looger LL, von Wirén N, Frommer WB. A cytosolic trans-activation domain essential for ammonium uptake. Nature. 2007 Mar 8;446(7132):195-8. doi: 10.1038/nature05579. PubMed 17293878 (Link opens in a new window)
- Sheff MA, Thorn KS. Optimized cassettes for fluorescent protein tagging in Saccharomyces cerevisiae. Yeast. 2004 Jun;21(8):661-70. doi: 10.1002/yea.1130. PubMed 15197731 (Link opens in a new window)
- Vickers CE, Bydder SF, Zhou Y, Nielsen LK. Dual gene expression cassette vectors with antibiotic selection markers for engineering in Saccharomyces cerevisiae. Microb Cell Fact. 2013 Oct 25;12:96. doi: 10.1186/1475-2859-12-96. PubMed 24161108 (Link opens in a new window)