Composite

Part:BBa_K2928014:Design

Designed by: Lauren Hartz   Group: iGEM19_ASIJ_Tokyo   (2019-10-16)


Modified 0.19 Wheat Albumin (Ile to Asn) expression using TDH3, RBS, secretion tag, and ADH3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We had to consider the strength of the promoters and terminators, with our intent being to maximize secretion and protein production while allowing the cells to have a relatively long lifespan. After experimentation, the above promoter and terminator were selected. Additionally, the choice to include a secretion tag is significant because we also created a variant of the construct without a secretion tag. The intent in designing this construct with a secretion tag was that it would aid in future protein purification, and potentially minimize the need to lyse the cells before analysis of the activity of the synthesized proteins.



Source

Excluding the 0.19 protein sequence, the parts composing this construct are all from previously existing parts in the iGEM parts registry. The original, unmodified 0.19 sequence was sourced from the Protein Database. We then modified it to our specifications.


References