Part:BBa_K1149051

Hybrid promoter phaCAB

In R. eutropha cells, P(3HB) is made through 3 steps. Two acetyl-CoA molecules made from carbohydrate converted to acetoacetyl-CoA by acetyl-CoA acetyltransferase encoded by PhaA gene. Then acetoacetyl-CoA reductase encoded by PhaB gene catalyzes the reduction of acetoacetyl-CoA to (R)-3-hydroxybutyryl-CoA (3HB-CoA). Finally, P(3HB) synthase encoded by PhaC gene catalyzes polymerization reaction of monomer molecules to polymer P(3HB) [2].

We cloned the constitutive promoter J23104+RBS B0034 upstream of the native phaCAB promoter and the phaCAB operon. Thus this part conssits of a hybrid promoter system.

References

1. H.Ohara. Change from Oil-based to Bio-based. Sen’i gakkaishi 66, 4.129-132 (2010) 2. Pohlmann, A. et al. Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16. Nature biotechnology 24, 1257–62 (2006).

J23104+RBS B0034-

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 979
  Illegal BglII site found at 1804
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 285
  Illegal NgoMIV site found at 356
  Illegal NgoMIV site found at 956
  Illegal NgoMIV site found at 1268
  Illegal NgoMIV site found at 1547
  Illegal NgoMIV site found at 2199
  Illegal NgoMIV site found at 2221
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4065