Device
Part:BBa_K3629013:Design
Designed by: Sravya Kakumanu Group: iGEM20_Calgary (2020-10-18)
Modified PfCBHI expression construct with gibson homology sequences and 6X His tag
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 2725
Illegal EcoRI site found at 2546 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal EcoRI site found at 2546
Illegal NheI site found at 75
Illegal SpeI site found at 2726
Illegal PstI site found at 2740
Illegal NotI site found at 7
Illegal NotI site found at 2733 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
Illegal EcoRI site found at 2546
Illegal BglII site found at 1322
Illegal BamHI site found at 2631
Illegal XhoI site found at 87 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 2726
Illegal EcoRI site found at 2546 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal EcoRI site found at 2546
Illegal XbaI site found at 16
Illegal SpeI site found at 2726
Illegal PstI site found at 2740
Illegal NgoMIV site found at 804 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Designed for Gibson Assembly
Source
The coding sequence of the CBHI is from Penicillium funiculosum, however it has been codon-optimized and mutated to have optimal activity in Yarrowia lipolytica based on our modelling. The promoter and terminator sequences are from wild-type Y. lipolytica.