Coding

Part:BBa_K3021002:Design

Designed by: Jeremy Hu   Group: iGEM19_PuiChing_Macau   (2019-10-14)
Revision as of 00:07, 15 October 2019 by Yexin003 (Talk | contribs)


NSP4-Laccase-HisTag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 92
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 930
    Illegal BglII site found at 1362
    Illegal BamHI site found at 1739
    Illegal XhoI site found at 962
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 145
    Illegal NgoMIV site found at 1601
    Illegal AgeI site found at 329
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 884
    Illegal SapI.rc site found at 394


Design Notes

The original sequence from the paper is ATGAAAAAGATTACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCG with the Protein sequence is The n-region: MKKI(the positive net charge in the n-region: +2) The h-region: TAAAGLLLLA The c-region: AQPAMA; We have codon optimized this sequence.


Source

Laccase sequence is codon optimized from previous iGEM team, which was initiated based on Trametes versicolor; and the secretion peptide, NSP4, is sequence from E.coli based from a previous publication.

References

References: Han, S., Machhi, S., Berge, M., Xi, G., Linke, T., & Schoner, R. (2017). Novel signal peptides improve the secretion of recombinant Staphylococcus aureus Alpha toxinH35L in Escherichia coli. AMB Express, 7, 93.