Part:BBa_K3841090
Expression cassette of soy leghemoglobin from methanol-dependent promoter
BBa_K3841090 is an expression cassette designed for Komagataella phaffii GS115 (previously referred to as Pichia pastoris). The soy leghemoglobin and TEF terminator are codon-optimized for K. phaffii under the control of the methanol-induced AOX1 promoter variant of Part:BBa_I764001.
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).
Assuming the AOX1 promoter variant has the same functionality as Part:BBa_I764001 presence of methanol will induce leghemoglobin production. Higher leghemoglobin yield is obtained when increasing the methanol concentration (see figure below).
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]
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