Part:BBa_K4579004

P_BAD promoter + RBS

This part includes the PTet promoter that, in the original pAJM Marionette plasmids, is repressed by tetR in the absence of anhydrotetracycline. This part can undergo digestion with BsaI to produce a linear sequence for the PTet promoter with sticky ends of 5' AACG 3' and 5' CATA 3'. This linear part can be combined with other parts in our chosen Golden Gate Assembly syntax (derived from the Bee Toolkit) to produce functional transcriptional units. In conjunction with BBa_K4579005, a part that includes the tetR regulatory gene, this part can be used as an inducible regulator of transcription. On its own, PTet acts constitutively, but the presence of the tetR repressor allows it to be induced by the addition of anhydrotetracycline (which removes the repressor from the promoter sequence).

The backbone for this part contains a gene encoding chloramphenicol acetyltransferase (cat) as a selective marker that imbues chloramphenicol resistance, and a high copy number ColE1 origin.

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 271
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 106
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 88