Part:BBa_K1632008

fim switch[default OFF](wild-type)_rbs_gfp

Fim switch (wild-type) is derived from wild-type sequence. Fim switch (wild-type) have sigma 70 promoter which is constitutively promoter. The promoter in the fim switch transcripts to the left. From the direction of transcription, the state is defined as OFF. On the other hand, fim switch [Default ON](wild-type)(BBa_K1632004) transcripts to the right.

Fim switch is inverted by two recombinase, FimB and FimE.The FimB protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction with approximately equal efficiencies. On the other hand, the FimE protein inverts fim switch predominantly in the ON-to-OFF direction.

We constructed fim switch [OFF](wild-type)-rbs-gfp(BBa_K1632008) to characterize the function of this part, by inserting fim switch [Default OFF](wild-type)(BBa_K1632005) upstream of a GFP coding sequence.

In the fimB dependent fim switch state assay, we transformed fim switch[OFF](wild-type)-rbs-gfp and pBAD/araC-fimB(BBa_K1632012)in the E.coli DH5alfa strain. We measured the fluorescence intensity of the cells induced by arabinose.

Similary,in the fimE dependent fim switch state assay, we transformed fim switch[OFF](wild-type)-rbs-gfp and pBAD/araC-fimE(BBa_K1632013) in the E.coli DH5alfa strain.We measured the fluorescence intensity of the cells induced by arabinose.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1052