Part:BBa_K3606108

P4 driven mcbABCD

BBa_K3606104 is one of a series of composite parts designed to screen a suitable strength promoter for mcbABCD.

Usage and Biology:

P4, which is a constitutive promoter, will continuously drive expression of mcbABCD. McbABCD is responsible for producing MccB17.

Design:

In our design the proper expression level of mcbABCD is very important. Since its product MccB17 acts as antibiotic, its low expression will make it unable to kill the surrounding bacteria and cannot improve its competitiveness in the intestine. But considering its toxicity, its high expression will also increase the burden of survival of engineered bacteria. So we need to test a collection of promoters, and find the most suitable one for it.

Further Application:

To use this part, simply clone it into a low-copy plasmid vector, transfer into E.coli, incubate overnight. You can also understand the strength of P4 through my experimental results.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 2327
  Illegal PstI site found at 2360
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 2327
  Illegal PstI site found at 2360
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 2327
  Illegal PstI site found at 2360
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 2327
  Illegal PstI site found at 2360
  Illegal NgoMIV site found at 1989
  Illegal AgeI site found at 2161
  
• 1000
  COMPATIBLE WITH RFC[1000]