Composite

Part:BBa_K3746007:Design

Designed by: Cheung Chi Ming   Group: iGEM21_Hong_Kong_JSS   (2021-10-18)


T7-LacO-DsbAss-tvLac-6His


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1643
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
    Illegal AgeI site found at 227
    Illegal AgeI site found at 1040
    Illegal AgeI site found at 1470
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The start codon of basic part tvLac (BBa_K3746002) is removed in our designed DNA due to a N-terminal signal peptide (DsbAss) is added into this secretory construct.

Source

The sequence of Trametes versicolor isolate K4 laccase CDS was extracted from NCBI (GenBank KR492189.1). Codon optimization for E. coli done by Thermo Scientific GeneArt platform. The other basic part sequences were identified from the iGEM part registry. The T7 promoter, LacO, RBS, and T7 terminator parts were used in the previous iGEM projects of our team. We plan to synthesize this composite part by IDT in form of gene fragments and clone it into the pSB1C3 vector by restriction cloning.

References