Part:BBa_K3196032

pGAP-Kozak-PHO1-pelA-His tag-TT-Pgpda-Kozak-FLO10-SLAC-His tag-TT-Pgas-Alkali-TT

We combine pelA, SLAC genes, which each adds His tag behind together.Through the previous experiment, we find the signal peptide which is most suitable for the target genes, and form the final pathway.Besides we add an alkali peptide to maintain pH.

Figure1 path way

Characterization

This is a three section part for degrading banana fiber part.

Usage and Biology

We put pectinase pelA at the first place because the banana fiber is covered by pectin. After dealing with the surface of the fiber, SLAC is as follow to degrade lignin inside the banana further, aiming to gain higher quality of the fiber. Since all these three enzymes’ optimum pH is near 5, we designed a pH system. We use Pgas, a promoter activate at pH equals 5, matching the optimum pH of the enzymes. In case the whole system’s pH decreases too much, we use Pgpda, a promoter activate at pH equals 2, connecting it with an alkali peptide to stabilize pH.

Figure 2

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1308
  Illegal BamHI site found at 478
  Illegal BamHI site found at 2127
  Illegal BamHI site found at 2138
  Illegal BamHI site found at 2149
  Illegal BamHI site found at 3501
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2588
  Illegal SapI site found at 238