Part:BBa_K1369002

Activator-Repressor promoter (AraC and TetR operator sites)

This is an activator-repressor (AR) combinatorial promoter which consists of an AraC operator site upstream of the -35 RNA polymerase box, and two TetR operator sites, one within the -35 and -10 RNA polymerase boxes, and the other downstream of the -10 RNA polymerase box. This AR promoter acts as a single-input gate (SIG): promoter activity is dependent only on aTc concentration.

In order to test the expression logic of this promoter, we designed test constructs with super-folder GFP, regulated by this AR 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]