Coding
Part:BBa_K4825011:Design
Designed by: Ni Weizhao Group: iGEM23_GreatBay-SCIE (2023-10-11)
LEU4^S547∆
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 397
Illegal XbaI site found at 511
Illegal SpeI site found at 552 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 75
Illegal SpeI site found at 552 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1282
Illegal BamHI site found at 228
Illegal BamHI site found at 1194 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 397
Illegal XbaI site found at 511
Illegal SpeI site found at 552 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 397
Illegal XbaI site found at 511
Illegal SpeI site found at 552 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We engineered LEU1, LEU2, and LEU4 with three different signal peptide, which all lead them into mitochondria. Therefore, the concentration of enzymes that involve in 2-ketoacid elongation would become much higher in mitochondria, so that KIV will be convert to KIC before encountering KDC. In the end, KIC would be secreted out into cytosol and convert to Isopentanol
Source
Saccharomyces cerevisiae