Coding

Part:BBa_K3924020

Designed by: Wenda Zhang   Group: iGEM21_Tsinghua   (2021-09-30)
Revision as of 07:40, 21 October 2021 by Songwh19 (Talk | contribs)

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mcmK. Modify MccH47 precursor


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]

Profile

Name: mcmK
Base Pairs: 1272bp
Origin: Escherichia coli, synthesis
Properties: Modify MccH47 precursor.

Usage and Biology

mcmK was obtained from E. coli strain CA46. It encodes McmK which modifies MccH47 precursor.[1]

Design and Construction

We introduce mcmK gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mcmK gene is added into lac operator(on RGP plasmid) so that the mcmK can only be expressed induced by IPTG.Due to the time cost of obtaining the sequence, the sequence was submitted to the company for synthesis and we sequenced it to verify the correctness of the synthetic sequence. The sequence was added to RGP plasmid by Hifi DNA assembly replacing the TMAR sequence for further use.

Figure 1: The design of mcmK


Reference

[1] Vassiliadis G, Destoumieux-Garzón D, Lombard C, Rebuffat S, Peduzzi J. Isolation and characterization of two members of the siderophore-microcin family, microcins M and H47. Antimicrobial Agents and Chemotherapy. 2010 Jan;54(1):288-297. DOI: 10.1128/aac.00744-09. PMID: 19884380; PMCID: PMC2798501.



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