Team:Kyoto/Parts

Parts

Basic Part

Our best basic part is BBa_K3815000. This is composed of ELK16, PT linker, Mxe Gyr intein. It can be expressed by attaching the sequence of the targeted protein to the N-terminus of Intein. This part is used for the purification method called ELK16system, which past iGEM teams had not been used. In this ELK16system, the ELK16 part is self-condensing, so the fusion protein can be extracted only by centrifugation, and the intein part is self-cleaved by DTT, so the target protein can be easily separated. The ELK16system will help improve the technology of recombinant protein purification, although it has some problems such as the presence of many other impurities in the protein recovery process.

Fig1. The mechanism of ELK16 system

Basic Part List

Name	Type	Part Name	Designer
BBa_K3815000	Protein Coding	ELK16-PT Linker-Mxe Gyr intein	Koudai Shiotsu
BBa_K3815001	Protein Coding	ELK16-PT Linker-Mxe Gyr intein-CecropinA	Koudai Shiotsu
BBa_K3815002	Protein Coding	ELK16-PT Linker-Mxe Gyr intein-Defensin1	Koudai Shiotsu
BBa_K3815003	Protein Coding	ELK16-PT Linker-Mxe Gyr intein-LL37	Koudai Shiotsu
BBa_K3815004	Protein Coding	ELK16-PT Linker-Mxe Gyr intein-NOP1	Koudai Shiotsu
BBa_K3815005	Protein Coding	ELK16-PT Linker-Mxe Gyr intein-NOP1v	Koudai Shiotsu
BBa_K3815006	Protein Coding	His tag-PT Linker-Mxe Gyr intein-CecropinA	Hiroto Koga
BBa_K3815007	Protein Coding	His tag-PT Linker-Mxe Gyr intein-Defensin1	Hiroto Koga
BBa_K3815008	Protein Coding	His tag-PT Linker-Mxe Gyr intein-LL37	Hiroto Koga
BBa_K3815009	Protein Coding	His tag-PT Linker-Mxe Gyr intein-NOP1	Hiroto Koga
BBa_K3815010	Protein Coding	His tag-PT Linker-Mxe Gyr intein-NOP1v	Hiroto Koga
BBa_K3815011	RNA	V-ATPaseB	Yuto Ueda
BBa_K3815012	RNA	TrxZ -3	Yuto Ueda
BBa_K3815013	RNA	EPH1	Yuto Ueda
BBa_K3815014	Composite	iRFP 670-superfolder GFP	Yuto Ueda
BBa_K3815032	Protein Coding	B. stearothermophilus DNA polymerase	Paper
BBa_K3815033	Protein Coding	HIV-1 reverse transcriptase p51	Paper
BBa_K3815034	Protein Coding	HIV-1 reverse transcriptase p66	Paper

Improvement of an Existing Part

These parts are the improvement from BBa_K1051206, the wildtype ssrA tag. Three C-terminal amino acids LAA in WT tag are replaced in these mutants, therefore resulting in a reduced protein degradation efficiency to various degrees (see Result). The collection of these mutants consist of a large repertoire of protein degradation tags of various levels, which can be applied for fine-tuning of many kinds of synthetic systems that require a precise control of proteins.

Improvement of an Existing Part List

Name	Type	Part Name	Designer
BBa_K3815015	Tag	AANDENYALGA.mutant SsrA degradation tag	Alexander Liu
BBa_K3815016	Tag	AANDENYAKWA.mutant SsrA degradation tag	Alexander Liu
BBa_K3815017	Tag	AANDENYALGALAK.mutant SsrA degradation tag	Alexander Liu
BBa_K3815018	Tag	AANDENYALGAHPK.mutant SsrA degradation tag	Alexander Liu
BBa_K3815019	Tag	AANDENYALGALAL.mutant SsrA degradation tag	Alexander Liu
BBa_K3815020	Tag	AANDENYALGANDK.mutant SsrA degradation tag	Alexander Liu
BBa_K3815021	Tag	AANDENYALGANEN.mutant SsrA degradation tag	Alexander Liu
BBa_K3815022	Tag	AANDENYALGAYES.mutant SsrA degradation tag	Alexander Liu
BBa_K3815023	Tag	AANDENYALGAHAK.mutant SsrA degradation tag	Alexander Liu
BBa_K3815024	Tag	AANDENYALGALAS.mutant SsrA degradation tag	Alexander Liu
BBa_K3815025	Tag	AANDENYALGATAP.mutant SsrA degradation tag	Alexander Liu
BBa_K3815026	Tag	AANDENYALGANAA.mutant SsrA degradation tag	Alexander Liu
BBa_K3815027	Tag	AANDENYALGAKLA.mutant SsrA degradation tag	Alexander Liu
BBa_K3815028	Tag	AANDENYALGALPA.mutant SsrA degradation tag	Alexander Liu
BBa_K3815029	Tag	AANDENYALGALAM.mutant SsrA degradation tag	Alexander Liu
BBa_K3815030	Tag	AANDENYALGALAI.mutant SsrA degradation tag	Alexander Liu
BBa_K3815031	Tag	AANDENYALGAPHA.mutant SsrA degradation tag	Alexander Liu