PARTS

Part collection

Vilnius-Lithuania iGEM 2024 project, Synhesion, focuses on developing biodegradable and environmentally sustainable adhesives with diverse applications. By leveraging the adhesive properties of bacterial polysaccharides and optimizing their production through engineered Escherichia coli, this project aims to provide innovative solutions that meet the growing need for eco-friendly and high-performance adhesives.

The project draws inspiration from two bacteria, Caulobacter crescentus and Hirschia baltica, which naturally produce adhesive polysaccharides that make up a structure called holdfast. Both aquatic organisms utilize a 12-protein synthesis pathway to produce these polysaccharides. By transferring this polysaccharide synthesis pathway into the industrially relevant E. coli strains (parts: BBa_K5246043, BBa_K5246046), we aim to develop a novel method for producing biodegradable adhesives on a larger scale.

Furthermore, the project establishes a foundation for future research by characterizing and purifying pathway proteins responsible for glycolipid production. This in particular would aid in liposome, with customizable oligosaccharide chains, creation opening up a wide range of applications in biomedicine. The BBa_K5246025, BBa_K5246026, BBa_K5246027, BBa_K5246028, BBa_K5246029 purified parts make up a system of tetrad assembly proteins integral to the holdfast synthesis pathway. This transferable system has the potential to produce specific glycolipid markers, functioning as molecular labels for targeted cell recognition and entry. These advancements hold significant promise in the fields of targeted drug and gene-editing system delivery, contributing to the rapidly expanding area of personalized medicine, which is poised to become a critical component of future healthcare solutions.

Our project’s part collection addresses pressing environmental concerns and hopes to improve the quality of life for people with diabetes. It also lays the groundwork for novel biotechnological applications in healthcare. We make a substantial contribution to the future of medicine and the development of sustainable medical materials.

Other teams are welcome to freely use our designed part collection to expand knowledge in these fields. By building on the foundation we have established, they can further characterize and optimize holdfast biosynthesis, and its applications or use individual proteins for custom glycolipid production, useful for liposome labeling for usage in medicine.

No.	Type	Part Number	Name	Description	New/existing
1	Basic	BBa_K5246025	CB2/CB2A HfsG, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
2	Basic	BBa_K5246026	CB2/CB2A HfsH, 6xHis tag	Deacetylase, 6xHis tag for purification	New
3	Basic	BBa_K5246027	CB2/CB2A HfsJ, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
4	Basic	BBa_K5246028	CB2/CB2A HfsK, 6xHis tag	Acetyltransferase, 6xHis tag for purification	New
5	Basic	BBa_K5246029	CB2/CB2A HfsL, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
6	Composite	BBa_K5246043	C. crescentus CB2/CB2A HfsE-HfsJ-HfsG-HfsL-HfsH-HfsK	Polysaccharide tetrad assembly system	New
7	Composite	BBa_K5246046	C. crescentus CB2/CB2A HfsA-HfsB-HfsD-HfsF-HfsC-HfsI	Polysaccharide export apparatus	New

Basic parts

No.	Type	Part Number	Name	Description	New/existing
1	Basic	BBa_K5246001	CB2/CB2A HfsA	Part of the export protein complex	New
2	Basic	BBa_K5246002	CB2/CB2A HfsB	Part of the export protein complex	New
3	Basic	BBa_K5246003	CB2/CB2A HfsC	Polysaccharide polymerase	New
4	Basic	BBa_K5246004	CB2/CB2A HfsD	Part of the export protein complex	New
5	Basic	BBa_K5246005	CB2/CB2A HfsE	Integral membrane glycosyltransferases	New
6	Basic	BBa_K5246006	CB2/CB2A HfsF	Flippase	New
7	Basic	BBa_K5246007	CB2/CB2A HfsG	Glycosyltransferase	New
8	Basic	BBa_K5246008	CB2/CB2A HfsH	Deacetylase	New
9	Basic	BBa_K5246009	CB2/CB2A HfsI	Polysaccharide polymerase	New
10	Basic	BBa_K5246010	CB2/CB2A HfsJ	Glycosyltransferase	New
11	Basic	BBa_K5246011	CB2/CB2A HfsK	Acetyltransferase	New
12	Basic	BBa_K5246012	CB2/CB2A HfsL	Glycosyltransferase	New
13	Basic	BBa_K5246013	HB HfsA	Part of the export protein complex	New
14	Basic	BBa_K5246014	HB HfsB	Part of the export protein complex	New
15	Basic	BBa_K5246015	HB HfsC	Polysaccharide polymerase	New
16	Basic	BBa_K5246016	HB HfsD	Part of the export protein complex	New
17	Basic	BBa_K5246017	HB HfsE	Integral membrane glycosyltransferases	New
18	Basic	BBa_K5246018	HB HfsF	Flippase	New
19	Basic	BBa_K5246019	HB HfsG	Glycosyltransferase	New
20	Basic	BBa_K5246020	HB HfsH	Deacetylase	New
21	Basic	BBa_K5246021	HB HfsI	Polysaccharide polymerase	New
22	Basic	BBa_K5246022	HB HfsJ	Glycosyltransferase	New
23	Basic	BBa_K5246023	HB HfsK	Acetyltransferase	New
24	Basic	BBa_K5246024	HB HfsL	Glycosyltransferase	New
25	Basic	BBa_K5246025	CB2/CB2A HfsG, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
26	Basic	BBa_K5246026	CB2/CB2A HfsH, 6xHis tag	Deacetylase, 6xHis tag for purification	New
27	Basic	BBa_K5246027	CB2/CB2A HfsJ, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
28	Basic	BBa_K5246028	CB2/CB2A HfsK, 6xHis tag	Acetyltransferase, 6xHis tag for purification	New
29	Basic	BBa_K5246029	CB2/CB2A HfsL, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
30	Basic	BBa_K5246030	HB HfsG, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
31	Basic	BBa_K5246031	HB HfsH, 6xHis tag	Deacetylase, 6xHis tag for purification	New
32	Basic	BBa_K5246032	HB HfsJ, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
33	Basic	BBa_K5246033	HB HfsK, 6xHis tag	Acetyltransferase, 6xHis tag for purification	New
34	Basic	BBa_K5246034	HB HfsL, 6xHis tag	Glycosyltransferase, 6xHis tag for purification	New
35	Basic	BBa_K5246035	BL21(DE3) ΔWecA	BL21(DE3) strain with WecA gene deletion	New
36	Basic	BBa_K5246036	Rosetta(DE3)pLysS ΔWecA	Rosetta(DE3)pLysS strain with WecA gene deletion	New
37	Basic	BBa_K5246037	HMS147(DE3) ΔWecA	HMS147(DE3) strain with WecA gene deletion	New
38	Basic	BBa_K5246038	BL21(DE3) ΔWecB	BL21(DE3) strain with WecB gene deletion	New
39	Basic	BBa_K5246039	Rosetta(DE3)pLysS ΔWecB	Rosetta(DE3)pLysS strain with WecB gene deletion	New
40	Basic	BBa_K5246040	HMS147(DE3) ΔWecB	HMS147(DE3) strain with WecB gene deletion	New
41	Basic	BBa_K5246054	dsDNA wecA No. 1	Double-stranded DNA for wecA gene impairment No. 1	New
42	Basic	BBa_K5246055	dsDNA wecA No. 2	Double-stranded DNA for wecA gene impairment No. 2	New
43	Basic	BBa_K5246056	dsDNA wecG	Double-stranded DNA for wecG gene impairment	New
44	Basic	BBa_K5246057	dsDNA wecF	Double-stranded DNA for wecF gene impairment	New
45	Basic	BBa_K5246058	dsDNA wzyE	Double-stranded DNA for wzyE gene impairment	New
46	Basic	BBa_K5246059	dsDNA wzzE	Double-stranded DNA for wzzE gene impairment	New
47	Basic	BBa_K5246060	pTargetF-_wecA_ No. 1	Guide RNA seed region for targeting wecA gene No. 1	New
48	Basic	BBa_K5246061	pTargetF-_wecA_ No. 2	Guide RNA seed region for targeting wecA gene No. 2	New
49	Basic	BBa_K5246062	pTargetF-_wecG_	Guide RNA seed region for targeting wecG gene	New
50	Basic	BBa_K5246063	pTargetF-_wecF_	Guide RNA seed region for targeting wecF gene	New
51	Basic	BBa_K5246064	pTargetF-_wzyE_	Guide RNA seed region for targeting wzyE gene	New
52	Basic	BBa_K5246065	Cas9 gRNA scaffold	Guide RNA scaffold for fully functional gRNA targeting specific genes	New
53	Basic	BBa_J64997	T7 promoter	Promoter induced by IPTG	Existing
54	Basic	BBa_K4165062	Lac Operator	Lac operator in plasmid regulated by IPTG	Existing
55	Basic	BBa_K2621038	RBS	Ribosome binding site	Existing
56	Basic	BBa_K731721	T7 terminator	Terminator under the control of T7 promoter	Existing

Composite parts

No.	Type	Part Number	Name	Description	New/existing
1	Composite	BBa_K5246041	C. crescentus CB2/CB2A HfsE-HfsJ-HfsG	Part of the polysaccharide tetrad assembly system	New
2	Composite	BBa_K5246042	C. crescentus CB2/CB2A HfsL-HfsH-HfsK	Part of the polysaccharide tetrad assembly system	New
3	Composite	BBa_K5246043	C. crescentus CB2/CB2A HfsE-HfsJ-HfsG-HfsL-HfsH-HfsK	Polysaccharide tetrad assembly system	New
4	Composite	BBa_K5246044	C. crescentus CB2/CB2A HfsA-HfsB-HfsD	Part of the polysaccharide export apparatus	New
5	Composite	BBa_K5246045	C. crescentus CB2/CB2A HfsF-HfsC-HfsI	Part of the polysaccharide export apparatus	New
6	Composite	BBa_K5246046	C. crescentus CB2/CB2A HfsA-HfsB-HfsD-HfsF-HfsC-HfsI	Polysaccharide export apparatus	New
7	Composite	BBa_K5246047	H. baltica HB HfsE-HfsG-HfsH	Part of the polysaccharide tetrad assembly system	New
8	Composite	BBa_K5246048	H. baltica HB CB2/CB2A HfsJ-HfsL-HfsK	Part of the polysaccharide tetrad assembly system	New
9	Composite	BBa_K5246049	H. baltica HB CB2/CB2A HfsE-HfsG-HfsH-HfsJ-HfsL-HfsK	Polysaccharide tetrad assembly system	New
10	Composite	BBa_K5246050	H. baltica HB CB2/CB2A HfsA-HfsB-HfsD	Part of the polysaccharide export apparatus	New
11	Composite	BBa_K5246051	H. baltica HB CB2/CB2A HfsF-HfsC-HfsI	Part of the polysaccharide export apparatus	New
12	Composite	BBa_K5246052	H. baltica HB CB2/CB2A HfsA-HfsB-HfsD-HfsF-HfsC-HfsI	Polysaccharide export apparatus	New
13	Composite	BBa_K5246066	gRNA for wecA gene editing with CRISPR/Cas9 No. 1	Fully functional gRNA targeting wecA No. 1	New
14	Composite	BBa_K5246067	gRNA for wecA gene editing with CRISPR/Cas9 No. 2	Fully functional gRNA targeting wecA No. 2	New
15	Composite	BBa_K5246068	gRNA for wecG gene editing with CRISPR/Cas9	Fully functional gRNA targeting wecG	New
16	Composite	BBa_K5246069	gRNA for wecF gene editing with CRISPR/Cas9	Fully functional gRNA targeting wecF	New
17	Composite	BBa_K5246070	gRNA for wzyE gene editing with CRISPR/Cas9	Fully functional gRNA targeting wzyE	New