Part:BBa_K4212038:Design
ChiS5
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1723
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1723
Illegal NheI site found at 1011 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1723
Illegal BglII site found at 1066
Illegal BamHI site found at 1708 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1723
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1723
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part consists the fusion protein consisted of CotZ and ChiS, in which we combined these parts into level 1 Golden Gate Construct. Our dry lab team predicted the functional model of this chimera and our wet lab intended to test the effectiveness of this model. We intended to compare the fungal-killing efficiency of CotZ and CotG fusion protein. Promoter hyper-spank is an inducible promoter, which was designed for high-level protein expression in B.subtilis and it was previously used by other iGEM teams.Our dry lab team predicted 2 possible chimeras, with either CotG or CotZ attached to the chitinase. Our wet lab team attempted to verify that the CotG chimera works better than the CotZ one. Promoter hyper-spank is an inducible promoter, which was designed for high-level protein expression in B.subtilis and it was previously used by other iGEM teams. The addition of Isopropyl β-D-1-thiogalactopyranoside (IPTG) can induce gene expression under the control of the promoter hyperspank. tL3S2P21 is a strong terminator, which helps to increase the production of desired protein.
Source
Synthetic construct.