Coding
Part:BBa_K4423001:Design
Designed by: Julia Vogl Group: iGEM22_BOKU-Vienna (2022-10-10)
Coding sequence for the main strain spider silk protein.
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 227
Illegal PstI site found at 422
Illegal PstI site found at 434
Illegal PstI site found at 581
Illegal PstI site found at 776
Illegal PstI site found at 788 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 227
Illegal PstI site found at 422
Illegal PstI site found at 434
Illegal PstI site found at 581
Illegal PstI site found at 776
Illegal PstI site found at 788 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 227
Illegal PstI site found at 422
Illegal PstI site found at 434
Illegal PstI site found at 581
Illegal PstI site found at 776
Illegal PstI site found at 788 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 227
Illegal PstI site found at 422
Illegal PstI site found at 434
Illegal PstI site found at 581
Illegal PstI site found at 776
Illegal PstI site found at 788
Illegal NgoMIV site found at 145 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sequence entails lots of repeating sections, which had to be considered during the codon optimization. Furthermore it is not possible to synthesize the whole sequence at once, it was devided in several parts that were connected through multiple PCR reactions.
Source
The sequence used was taken from "Bin Liu, Tao Wang, Liyan Xiao, Guilan Zhang, Guangshen Li, Jingzhi Luo & Xiaobing Liu (2018) A directed self-assembly quasi-spider silk protein expressed in Pichia pastoris, Biotechnology & Biotechnological Equipment, 32:2, 451-461, DOI: 10.1080/13102818.2017.1327823" and later codonoptimized using the codon optimization tool of the company IDT.