Part:BBa_K3924021

mcmL

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 362
  Illegal PstI site found at 1067
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 362
  Illegal PstI site found at 1067
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 362
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 362
  Illegal PstI site found at 1067
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 362
  Illegal PstI site found at 1067
  Illegal AgeI site found at 537
  
• 1000
  COMPATIBLE WITH RFC[1000]

Profile

Name: mcmL
Base Pairs: 1119bp
Origin: Escherichia coli, synthesis
Properties: Modify MccH47 precursor

Usage and Biology

mcmL was obtained from E. coli strain CA46. It encodes McmL which modifies MccH47 precursor.^[1]

Design and Construction

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

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.