Composite

Part:BBa_K3841074

Designed by: Emil Ørsted Christensen   Group: iGEM21_DTU-Denmark   (2021-10-17)


Expression cassette of pMMO alpha subunit fused to Venus with constitutive promoter

BBa_K3841074 is an expression cassette designed for Komagataella phaffii GS115. The PmoB protein has been fused with a signal peptide, that is predicted to transport the construct to the plasma membrane of K. phaffii GS115, and a flourescent protein, Venus. The PmoB protein, the Venus protein and the TEF terminator are codon-optimized for K. phaffii under the control of the TEF1 promoter.

Folding of PmoB fused to Venus(BBa_K3841015) with signal peptide (BBa_K3841036) and PmoB(BBa_K3841010) simulated using AlphaFold [1]. The coding sequence in this expression cassette follows the lime structure.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1070
    Illegal XbaI site found at 1604
    Illegal PstI site found at 1925
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1925
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 785
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1070
    Illegal XbaI site found at 1604
    Illegal PstI site found at 1925
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1070
    Illegal XbaI site found at 1604
    Illegal PstI site found at 1925
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Cramer, Patrick. “AlphaFold2 And the Future of Structural Biology.†Nature Structural and Molecular Biology, vol. 28, no. 9, Nature Research, 2021, pp. 704–05, doi:10.1038/s41594-021-00650-1.

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