Composite

Part:BBa_K3606060:Design

Designed by: Yuanyuan Liu   Group: iGEM20_Fudan   (2020-10-23)


P4 driven mcbABCD and PtetR driven mcbEFG


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5548
    Illegal XbaI site found at 5533
    Illegal PstI site found at 2371
    Illegal PstI site found at 2404
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5548
    Illegal PstI site found at 2371
    Illegal PstI site found at 2404
    Illegal NotI site found at 5540
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5548
    Illegal BamHI site found at 3367
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5548
    Illegal XbaI site found at 5533
    Illegal PstI site found at 2371
    Illegal PstI site found at 2404
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5548
    Illegal XbaI site found at 5533
    Illegal PstI site found at 2371
    Illegal PstI site found at 2404
    Illegal NgoMIV site found at 2033
    Illegal AgeI site found at 2205
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

McbABCD and McbEFG are placed in reverse, and each is independently expressed under different promoters with their own terminator.


Source

McbABCD and McbEFG orginally come from a plasmid of Escherichia coli.

References

Collin F, Maxwell A. The Microbial Toxin Microcin B17: Prospects for the Development of New Antibacterial Agents. J Mol Biol. 2019;431(18):3400–3426. doi:10.1016/j.jmb.2019.05.050

S. Duquesne, D. Destoumieux-Garzón, J. Peduzzi, S. Rebuffat. Microcins, gene-encoded antibacterial peptides from enterobacteria

Sassone-Corsi M, Nuccio SP, Liu H, Hernandez D, Vu CT, Takahashi AA, Edwards RA, Raffatellu M. Microcins mediate competition among Enterobacteriaceae in the inflamed gut. Nature. 2016 Dec 8;540(7632):280-283.

Mutalik VK, Guimaraes JC, Cambray G, Lam C, Christoffersen MJ, Mai QA, Tran AB, Paull M, Keasling JD, Arkin AP, Endy D. Precise and reliable gene expression via standard transcription and translation initiation elements. Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 2013 Mar 10. PMID: 23474465.