Part:BBa_K1163999

Inverter composed of Fur Binding site from AceB promoter + LacI-LVA

Promoter from E. coli that controls the AceB gene, involved in iron uptake.

This promoter sequence contains a RBS and a FUR binding site, although it is not clearly possible to identify them precisely. It has been shown to downregulate the expression of sfGFP or any gene down-stream in the presence or iron in the range 10^-7, 10^-5 mol.L-1

FUR (Ferric Uptake Regulation) is a transcriptional repressor of genes involved in iron homeostasis. In presence of iron FUR will be activated and it modification of conformation will induce a dimerization of two FUR. When the FUR protein is dimerized, it will bindind to the DNA in a Fur Binding Site, then FUR will inhibit the mRNA transcription. To realize our project we want to use this Fur binding site merged with an inhibitor (here LacI) to create an inverter system.

AceB regulatory promoter will decrease the expression of the LacI-LVA gene (LacI protein with a LVA degredation tag) when the FUR protein is dimerized in the presence of iron. This part should be used to express a gene of interest that is under control of a Lac operator (LacO). This way, the gene will be positively regulated in the presence of iron.

This system has been adapted to B. subtilis by the UPMC-Paris Team in 2016. ( see part BBa K218004 )

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 1208
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]