Composite

Part:BBa_K5322007:Design

Designed by: Yingying Yu   Group: iGEM24_NJTech-China   (2024-10-01)
Revision as of 07:58, 1 October 2024 by Casey (Talk | contribs) (Design Notes)


NO-inducible Mfp53 Expression System


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 552
    Illegal NheI site found at 1113
    Illegal NotI site found at 1077
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 763
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To ensure that the animal-derived mussel foot protein Mfp53 can be successfully expressed in Escherichia coli Nissle 1917, we performed codon optimization on the Mfp53 sequence.


Source

The plasmid pET29a-J23119-SoxR-T-pSoxS-RBS-Mfp53-T7 utilizes the pET29a vector, which is typically used for high-level expression of genes in Escherichia coli. The components used in this design, including the J23119 promoter, Mfp3, Mfp5, the protein linker (GGGGS), SoxR protein, and SoxS promoter, are all sourced from the iGEM registry of standard biological parts (BBa_J23119BBa_K4854000BBa_K1583002,BBa_K5322001,BBa_K554003BBa_K5322004). Among these, the mussel foot protein Mfp5 and Mfp3 are derived from natural marine mussels. These components were selected for their validated functions and compatibility in synthetic biology applications, facilitating controlled expression of genes within bacterial cells under specific conditions.

References