Part:BBa_K2088672

copA + riboJ +mhbDHIM + terminator

The composite part contains copA promoter, riboJ , a gene cluster mhbDHIM , and a terminator. The promoter is induced by copper ion and transcribe downstream genes. The riboJ between copA and the gene cluster can decrease interference of one part type （promoter）on another（protein coding sequence）. The gene cluster mhbDHIM contains four connected genes which can degrade 3-hydroxybenzoate to the Fumarate and Pyruvate step by step.

After experienting, we found that the device can degrade the 3-hydroxybenzoate completely in short time. Also, engineering bacteria with the device grows better in the medium with 3-hydroxybenzoate than Klebsiella pneumoniae M5a1which is capable of utilizing 3-hydroxybenzoate via gentisate, and can tolerate higher concentration of 3- hydroxybenzoate.In conclusion, the device can help the E.coli degrade the 3- hydroxybenzoate effectively.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2518
  Illegal BamHI site found at 2904
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 480
  Illegal NgoMIV site found at 754
  Illegal NgoMIV site found at 2708
  Illegal NgoMIV site found at 3845
  Illegal AgeI site found at 816
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3038
  

Figure1:3-HBA was degraded thoroughly by our engineered E.coli transformed with this composite part.

Figure2:The ordinate is the concentration of 3-HBA. The left column shows the figure of Klebsiella pneumoniae M5a1, while the right one shows the figure of our engineered bacteria. From this figure we can find that our engineered bacteria can degrade 3-HBA much faster than M5a1 in 24h.