Part:BBa_K1369000

Repressor-Repressor promoter (TetR and LacI operator sites)

This is a repressor-repressor (RR) combinatorial promoter which consists of a TetR operator site within the -35 and -10 RNA polymerase boxes and a LacI operator site downstream of the -10 RNA polymerase box. This RR promoter behaves as an AND-gate, where the promoter is only active with both aTc and IPTG inducers are present (in sufficient concentrations).

In order to test the expression logic of this promoter, we designed test constructs with super-folder GFP, regulated by this RR promoter and a bicistronic RBS (BBa_J119024). A bicistronic RBS (BCD) was instead of a standard RBS because it has been shown that BCDs have more robust gene translation with respect to the downstream sequence.

Heatmap of mean GFP fluorescence (au) as regulated by this RR promoter in vivo. Cells were grown for around 8 hours with various inducer concentrations before measurements were taken

Barchart of mean GFP fluorescence (au) as regulated by this RR promoter in vivo. This barchart was obtained using the same data that generated the above heatmap.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]