Difference between revisions of "Part:BBa K3021002:Design"
 
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===Design Notes===
 
===Design Notes===
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The original NSP4 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 and also the Laccase sequence from k863030.
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Latest revision as of 00:08, 15 October 2019


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 NSP4 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 and also the Laccase sequence from k863030.


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.