Part:BBa_K1632000

fim switch[deault ON](Tokyo_Tech/J23119)

Fig. 1. fim switch(Tokyo_Tech/J23119) design (Up:[ON] state Down:[OFF] state)

The fim switch is the promoter containing repeated DNA sequence which is inverted by the Fim recombinase. Therefore, we can control the expression of the gene downstream of the fim switch by adding the Fim recombinase.
We designed this fim switch which has a J23119 promoter. Also between the promoter and the inverting site, there are two restriction enzyme sites in each front (SalIand BamHI) and back (BglII and MluI)(Fig. 1. fim switch(Tokyo_Tech/J23119) design). So the promoter can easily be interchanged. Except for insertion of restriction enzyme sites, basically, the design of fim switch(Tokyo_Tech) is similar with fim switch(wild-type). From our results of our assay, the inversion of fim switch(Tokyo_Tech/J23119) by FimB/FimE was not confirmed correctly. The FimB protein inverts fim switch from [ON] state to [OFF] state and from [OFF] state to [ON] state correctly. However the FimE protein inverts fim switch predominantly from [ON] state to [OFF] state. In the assay, the FimE(wild-type) protein inverts fim switch from [ON] state to [OFF] state and from [OFF] state to [ON] state. In other words, the FimE protein works as the FimB protein. Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 345
  Illegal NheI site found at 368
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 374
  Illegal BamHI site found at 333
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]