Part:BBa_K3841025
Expression cassette of yeast optimized Venus from methanol-dependent promoter
BBa_K3841025 is an expression cassette designed for Komagataella phaffii GS115 (previously referred to as Pichia pastoris. The Venus protein and TEF terminator are codon-optimized for K. phaffii under the control of the methanol-induced AOX1 promoter.
Usage
For chromosomal integration, flank the expression cassette by 65bp long homology arms. The homology arms should match upstream and downstream to an induced DSB. Visit 2021 DTU-Denmark’s Toolbox for K. phaffii to find homology arms. Insert the fragment into a vector backbone of choice for instance the pGGAselect destionation plasmid (BBa_K3841023).
The part is designed for measuring the AOX1 promoter strength in K. phaffii. Upon cultivation in methanol containing medium, the system is activated and the transciptional activity can be monitored using a flourescent filter for Venus. Venus has an excitation wavelength of 515 nm and emission wavelength of 528 nm [1].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
[1] Shaner, Nathan C., et al. “A Guide to Choosing Fluorescent Proteins.” Nature Methods, vol. 2, no. 12, NATURE PUBLISHING GROUP, 2005, pp. 905–09, doi:10.1038/nmeth819.
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