Part:BBa_K4769000

pMiniMAD : Backbone for markerless HR into B. subtilis

pMiniMAD is a high copy number plasmid containing ampicillin resistance (for selection in E. coli) and erythromycin resistance (for selection in Bacillus subtilis). In addition to the high copy number ColE1 ori, pMiniMAD contains the temperature-sensitive origin of replication, pE194ts, allowing for markerless homologous recombination into B. subtilis (see https://2023.igem.wiki/cambridge/protocols for details on this protocol)

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9
  Illegal XbaI site found at 1915
  Illegal XbaI site found at 6344
  Illegal PstI site found at 2391
  Illegal PstI site found at 6332
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9
  Illegal PstI site found at 2391
  Illegal PstI site found at 6332
  Illegal NotI site found at 378
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9
  Illegal BamHI site found at 6350
  Illegal XhoI site found at 4092
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9
  Illegal XbaI site found at 1915
  Illegal XbaI site found at 6344
  Illegal PstI site found at 2391
  Illegal PstI site found at 6332
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9
  Illegal XbaI site found at 1915
  Illegal XbaI site found at 6344
  Illegal PstI site found at 2391
  Illegal PstI site found at 6332
  Illegal AgeI site found at 3292
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 5001
  Illegal SapI.rc site found at 6083