Part:BBa_K3590036

Immobilization Module

The purpose of the Immobilization module (IM) is to immobilize the cells of B. subtilis on a cellulose matrix. This construct was designed to ensure that the genetically modified bacteria will not escape our floating device and endanger the environment.

At the 5' end of the IM there is a EcoRI restriction site and the Pveg promoter followed by the RBS R2. The coding sequence (CDS) encodes several functional modules - SacB signal sequence for extracellular transport, Myc-tag for Western blot detection, followed by the cellulose binding module (CBM) through which it will attach to the microbeads in our device. The CBM is connected by a linker to three copies of the LysM domains which anchor the whole system to peptidoglycans in the cell wall of B. subtilis. At the 3' end of the CDS, there is a STOP codon followed by the HindIII restriction site.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 1083
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 1083
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 170
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 1083
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 1083
  Illegal AgeI site found at 185
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1145