Part:BBa_K4618213:Design
Glac1+ NG catcher
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1764
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1764
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1764
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1764
Illegal AgeI site found at 996 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This sequence is combined with Glac and NGCacther. The function of Glac is degradation of textile dyes. And the function of NGCatcher is used to bind the tag which combine with other protein. This sequence is derived from Optimization of Laccase from Ganoderma lucidum Decolorizing Remazol Brilliant Blue R and Glac1 as Main Laccase-Contributing Gene and Tailoring the Tag/Catcher System by Integrating Covalent Bonds and Noncovalent Interactions for Highly Efficient Protein Self-Assembly. However, we did not directly employ the sequences documented in the literature. We revise the sequence to let it fit E.coli more. Besides, incorporating the existing Glac to remove the stop codon, eliminating the catcher's methionine, allowing the concatenation of both sequences into a single protein. Then we entrusted Twist Bioscience with its synthesis and subsequent cloning into a plasmid. We use this part in the biosafety level 1 laboratory. Because we used E.coli system to express this part, so we can’t promise it can be expressed in other system. Lastly, we employ SDS-PAGE to verify the accuracy of expressed protein's molecular weight.
Source
GenBank: KC507935.1
References
Paper:Optimization of Laccase from Ganoderma lucidum Decolorizing Remazol Brilliant Blue R and Glac1 as Main Laccase-Contributing Gene
Peng Qin,1,† Yuetong Wu,1,† Bilal Adil,1 Jie Wang,1 Yunfu Gu,1 Xiumei Yu,1 Ke Zhao,1 Xiaoping Zhang,1 Menggen Ma,1 Qiang Chen,1 Xiaoqiong Chen,2 Zongjin Zhang,3 and Quanju Xiang1,*Molecules. 2019 Nov; 24(21): 3914."