Part:BBa_K305011:Design

Subtilin inducible expression plasmid for Bacillus subtilis

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 3141
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 3147
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3141
  Illegal BglII site found at 3060
  Illegal XhoI site found at 287
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 3141
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 3141
  Plasmid lacks a suffix.
  Illegal XbaI site found at 3156
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal SapI site found at 324
  

Design Notes

Contains a BamHI restriction site in between the prefix and suffix.

Cloning with this plasmid in certain E. coli strains (MC1061 and TOP10) results in the insertion of an IS element of around 800bp containing a PstI site. It can be detected by a PstI restriction, yielding two fragments of 2783 and 1161bp's, or by a PCR using the following primers, producing a 525, or 1301bp product.

forward: ATTGAGAGGAGGGATTATTG reverse:GAGTTTATCACCCTTGTC

Source

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