Coding

Part:BBa_K4806001:Design

Designed by: Luca Langenberg   Group: iGEM23_RPTU-Kaiserslautern   (2023-09-13)


CYP2D6 gene for Chlamydomonas reinhardtii (Phytobrick)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 356
    Illegal PstI site found at 1712
    Illegal PstI site found at 2145
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 356
    Illegal PstI site found at 1712
    Illegal PstI site found at 2145
    Illegal NotI site found at 862
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 356
    Illegal PstI site found at 1712
    Illegal PstI site found at 2145
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 356
    Illegal PstI site found at 1712
    Illegal PstI site found at 2145
    Illegal NgoMIV site found at 999
    Illegal NgoMIV site found at 1348
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part has been codon optimized for Chlamydomonas reinhardtii and it is compatible with Phytobricks/MoClo parts.
We had to insert one mutation at 586 bp to fit to the MoClo standards of iGEM. In our lab we are not using the restriction enzyme SapI, but instead BbsI. Therefore, this mutation does not occure in the parts we used in our experiments. This mutation is silent, but it might affect the expression level, since the triplet AGT is much rarer in Chlamydomonas, compared to the triplet AGC, which we used.
Further, we had to incorporate introns because Chlamydomonas reinhardtii is an eukaryotic organism.

Source

https://www.uniprot.org/uniprotkb/P10635/entry

References