Reporter

Part:BBa_K3841088

Designed by: Karen Therkelsen   Group: iGEM21_DTU-Denmark   (2021-10-18)


Expression cassette of soy leghemoglobin from methanol-dependent promoter

BBa_K3841088 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 and can be expressed 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).

In presence of methanol, leghemoglobin is produced. Higher leghemoglobin yield is obtained when increasing the methanol concentration (see figure below).

Promoter kinetics model of AOX1 promoter for the production of soy leghemoglobin (BBa_K3841014) as a function of various methanol concentrations.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None