Part:BBa_K1597001

ThrC integration backbone for B. subtilis

This biobrick is a shuttle vector and can replicate in E. coli and integrate in the thrC locus from Bacillus subtilis .

During Bacillus subtilis transformation, the wanted DNA can get integrated in a loci present in the genomic DNA. One of these loci is the thrC locus. To have an extra integration point in B. subtilis this biobrick was developed. This backbone is based on the plasmids pdg1664 (AM Guérout-Fleury, 1996) and BBa_K823023 . The amyE locus front and back from BBa_K823023 were replaced with the thrC locus front and back from pdg1664. Along with this exchange, the chloramphenicol resistance gene was replaced with erythromycin resistant gene (wich was also found in pdg1664). This biobrick can be replicated both in Escherichia coli and in Bacillus subtilis, where it integrates into the genome.

This biobrick was successfully integrated in B. subtilis.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 5126
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 5132
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 5126
  Illegal BamHI site found at 5108
  Illegal XhoI site found at 2503
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 5126
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 5126
  Plasmid lacks a suffix.
  Illegal XbaI site found at 5141
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 2467
  Illegal NgoMIV site found at 3606
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI.rc site found at 3430