Composite

Part:BBa_K3834037

Designed by: HU Feijun   Group: iGEM21_CityU_HK   (2021-10-21)


sC(front)_PamyB_TamyB_sC(rear)

The composite biobrick (as shown in Figure 1) takes advantage of an sC locus to facilitate the efficient and specific integration of any “exogenous gene fragment” into the Aspergillus oryzae host chromosome. In addition, via CRISPR-Cas9-mediated homologous recombination (using a tailored gRNA homologous to the sC locus), transformation of this biobrick harboring any exogenous gene (e.g. Stl-P450 and acldA-P450) into the fungal host is highly efficient and requires significantly less plasmid DNA (Figure 2).

T--CityU_HK--parts4027.png

Figure 1. Genetic circuit of sC(front)_PamyB_(exogenous gene)_TamyB_sC(rear).


T--CityU_HK--parts4027-2.png

Figure 2.CRISPR-Cas9-mediated homologous recombination method requires significantly less plasmid DNA for efficient fungal transformation. (old = transformation protocol without Crispr step; new = transformation protocol with Crispr step).

Sequence and Features


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]


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Parameters
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