Composite

Part:BBa_K4968011:Design

Designed by: Shouye Zhu   Group: iGEM23_XJTLU-CHINA   (2023-10-09)

Optimized SUMO tag

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1958
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1958
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1958
    Illegal BglII site found at 198
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1958
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1958
    Illegal NgoMIV site found at 58
    Illegal AgeI site found at 2269
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Considering that JF1 knocks out the entire curli operon in the genome, CsgA-AG4-CsgC-CsgD-CsgE-CsgF-CsgG is needed in the plasmid. CsgA-AG4-CsgC-CsgD-CsgE-CsgF-CsgG includes both target fusion protein and assistance protein, which ensure the integrity of protein expression, secretion, and self-assembly. Since the complete curli operon was required on the plasmid, we chose to transfer the relevant sequences from the original bacterial genome to the plasmid. Each gene was linked to a sequence containing RBS, ensuring normal expression of each gene.


Source

The sequence of AG4 is from the literature “Solution structure of peptide AG4 used to form silver nanoparticles” (Asn1-Pro-Ser-Ser-Leu-Phe-Arg-Tyr-Leu-Pro-Ser-Asp). The sequence of CsgA, CsgC, CsgD, CsgE, CsgF, and CsgG are from NCBI.