Part:BBa_J64032

pCASP SPI-1 Secretion Circuit

The pCASP circuit is a protein expression and secretion circuit. The system is currently only functional when expressed in Salmonella under SPI-1 expressing conditions. When SPI-1 is expressed the pCASP circuit will be activated for expression at the same time natural effectors would normally turn on and secrete. This circuit will express a protein of interest fused to the SptP167 secretion signal along with co-expression of the sicP chaperone. This combination is sufficient to direct the protein of interest to the export machinery and elicit secretion to the extracellular environment. In the initial configuration of this circuit the DH guanine exchange factor is used as a de-bugging secreted protein.

Note: Some evidence exists of impassible substrates for the secretion machinery. Most of these proteins are highly thermodynamically stable domains that cannot be unfolded to transit through the secretion system.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 1767
  Illegal PstI site found at 533
  Illegal PstI site found at 1158
  Illegal PstI site found at 1388
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 533
  Illegal PstI site found at 1158
  Illegal PstI site found at 1388
  Illegal NotI site found at 1684
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1478
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 1767
  Illegal PstI site found at 533
  Illegal PstI site found at 1158
  Illegal PstI site found at 1388
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1767
  Illegal PstI site found at 533
  Illegal PstI site found at 1158
  Illegal PstI site found at 1388
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1632