Reporter

Part:BBa_K3841093

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


Expression cassette of mOrange from methanol-dependent promoter

BBa_K3841093 is an expression cassette designed for Komagataella phaffii GS115 (previously referred to as Pichia pastoris. The mOrange is the Part:BBa_E2050 from the distribution kit 2021. The TEF terminator is 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 mOrange.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1702
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None