Part:BBa_K2962000:Design
lcc1 gene codes laccase enzyme.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 779
Illegal BglII site found at 1211
Illegal XhoI site found at 811
Illegal XhoI site found at 1444 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1297
Illegal NgoMIV site found at 1450
Illegal AgeI site found at 178 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 733
Illegal SapI.rc site found at 243
Design Notes
To prevent the degeneration of codon optimization are done according to E.coli. To increase the level of activity and yield of enzyme designed are done which contains different sequences. These designs contain pelb sequence, dsb fusion tag, and histidine tag. pelb sequence is done for increasing the activity and the yield of the enzyme in the periplasmic region. dsb fusion tag is done for increasing the interaction of disulfide-bound and thus it is thought that the activity of the enzyme will increase. Finally, histidine tag is added to increase the yield of isolation.
Characterization
Please look at iGEM10_Tianjin team’s laccase from Trametes versicolor, we characterized their laccase by adding some advanced cells or peptides. The Trametes versicolor laccase gene (Laccase (lcc1) BBa_K2962000) is an optimized sequence for expression in the E. coli organism. Characterized by team iGEM10_Tianjin (BBa_K500002) To avoid codon degeneration, we innovated a new sequence. We designed all of our other parts according to this laccase.
Source
The source of the lcc1 gene will be obtained from its sequence on GenBank. Gene is available at accession number: Y18012.1.