Composite

Part:BBa_K4059032:Design

Designed by: Ann-Sofie Nielsen   Group: iGEM21_SDU-Denmark   (2021-10-21)


PsiT1, E. coli K12 codon optimized


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 473
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1093
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The biobrick is a theoretical transporters of psilocybin from P. cubensis genome. It contains the sequence of PsiT1 which is one of two theoretical transporters with the other being (BBa_K4059033).

Considerations: We used Swiss-Model for protein structure homology-modelling and found that Major facilitator-type transporter psiT1 and Major facilitator-type transporter psiT2 shows similarity with known ion coupled transporters. This indicated that the proteins are transporters which drove us to continue with experiments. Due to the theoretical nature of this biobrick it was designed for introduction of a BamHI restriction site by specific primers on either C-terminal or N-terminal. This is done to enable ligation to our biobricks with our GFP constructs BBa_K4059034 and BBa_K4059035. The sequence has been codon optimized for E. coli from the original DNA sequence and therefore modified slightly.

Source

https://pubmed.ncbi.nlm.nih.gov/28763571/

References