Part:BBa_K3196031
pGAP-Kozak-FLO10 pro-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.
Characterization
This is a three section for degrade 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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1527
Illegal BamHI site found at 478
Illegal BamHI site found at 2346
Illegal BamHI site found at 2357
Illegal BamHI site found at 2368
Illegal BamHI site found at 3720 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2807
Illegal SapI site found at 238
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