Part:BBa_K1772000:Design
yEGFP-tagged Mn peroxidase
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 346
Illegal PstI site found at 1018 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 346
Illegal PstI site found at 1018 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 788
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 346
Illegal PstI site found at 1018 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 346
Illegal PstI site found at 1018 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Our goal was to synthesize lignin-degrading enzymes to be secreted out of yeast cells, which we used the secretion tag for. The fluorescence tag was merely implemented to simplify detection of successful secretion. To avoid time-consuming and low-efficiency standard assembly techniques, this construct was designed to be synthesized by IDT using two GBlocks(c), which were to be joined by Gibson Assembly. Linker sequences were implemented in order to minimize interaction between protein domains. A constitutive promoter was implemented due to the intended application (a bioreactor pretreatment process where a yeast containing this part would secrete enzymes to continuously degrade lignin).
Source
Promoter: BBa_I766555 Kozak sequence: BBa_J63003 yEGFP (yeast Enhanced Green Fluorescent Protein): NCBI Genbank accession U73901.1 Linker sequences: BBa_K243004 Manganese Peroxidase from Heterobasidion irregulare TC 32-1, NCBI Reference Sequence: NW_009258207.1 Secretion tag: Bba_K416003 Terminator: BBa_K392003