Composite

Part:BBa_K4059042:Design

Designed by: Julie le-Fevre Hansen   Group: iGEM21_SDU-Denmark   (2021-10-21)


PsiH from Psilocybe cubensis with N-terminal substitution (5), E. coli K12 codon optimized, with Lac


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2125
    Illegal SapI.rc site found at 1747


Design Notes

The psiH gene encodes PsiH, a cytochrome p450 monooxygenase naturally found in Psilocybe cubensis (fungi). It is involved in the synthesis of Psilocybin, a psychedelic substanse produced from the amino acid tryptophane. The role of PsiH is to catalyze the conversion of tryptamine to 4-hydroxytryptamin. Wild type PsiH has a hydrofobic N-terminal domain, which is associated with the membrane of the endoplasmatic reticulum in the eucaryotic cells of P. cubensis. In order to express the PsiH-enzyme in procaryotes such as E. Coli, the iGEM SDU 2021 team have designed this biobrick, in which the psiH sequence is codon optimised for E. Coli. Furthermore, the different variants of the psiH-biobrick contain different modifications of the N-terminal encoding gene region. This specific biobrick contains an N-terminal substitution, where the entire N-terminal has been substituted with an N-terminal from a different protein, resulting in a chimeric protein.

Source

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References