Part:BBa_K3924025

mchC, function unknown

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 1017
  Illegal PstI site found at 126
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 126
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 417
  Illegal BglII site found at 580
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 1017
  Illegal PstI site found at 126
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1017
  Illegal PstI site found at 126
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 464
  Illegal SapI.rc site found at 1384
  

Profile

Name: mchC
Base Pairs: 1551bp
Origin: Escherichia coli, synthesis
Properties: May be microcin H47(MccH47) modification protein

Usage and Biology

mchC was obtained from E. coli strain CA46. It encodes MchC which might modifies the precursor of MccH47.^[1]

Design and Construction

We introduce mchC gene from E.coli CA46 into E.coli Nissle 1917. The sequence was obtained from E. coli CA46 microcin operon(GenBank: AJ515251.1). The mchC gene is added into lac operator(on RGP plasmid) so that the mchC 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 mchC

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.