Overview
Here one can find a collection of all the parts in the categories excisting-, basic-, and composite parts used in our experiments. The composite parts will contribute to the Registry of standard biological parts, expanding the resources for future iGEM teams.
Table of excisting parts
Table over existing parts with their corresponding name and description, used in our project.
| Parts registry ID | Type | Part name | Description |
|---|---|---|---|
| BBa_K143013 | Regulatory | Promoter 43 a constitutive promoter for Bacillus subtilis | Promoter 43 is a constitutive promoter that constitutively expresses the P43 protein in "B. subtilis". This promoter has been shown to be recognized and active during the exponential and lag phases of growth. It has been hypothesized that the ability to recognize the promoter in exponential and lag phase of growth is due to the recognition of the promoter by both sigma factor 55 (the major sigma factor) and sigma factor 37 (the lag phase sigma factor). |
Table of basic parts
Table over basic parts submitted with their corresponding name and description.
| Parts registry ID | Type | Part name | Description |
|---|---|---|---|
| BBa_25WLFEMG | Coding | CHS3 - native expression gene of chitinase in S. cerevisiae | CHS3 encodes the class III chitin synthase (Chs3p) in Saccharomyces cerevisiae. It is a multi-pass membrane enzyme that polymerizes chitin from UDP-GlcNAc and inserts it into the yeast cell wall. |
| BBa_25Q96I72 | Coding | CHI92 - Chitin cell wall binding domain | CHI92 is a chitinase-derived adhesin module displayed on the cell surface of E. coli Nissle to enable binding to the fungal cell wall (specifically Candida albicans) via chitin or related polysaccharide components. In engineered bacteria, Chi92 should be fused to a surface-anchoring domain so that it is exposed externally, where it is expected to bind fungal cell wall structures, promoting stable adhesion to the pathogen. Its activity does not rely on native adhesion factors of E. coli but uses the carbohydrate-binding affinity of the chitinase domain to mediate specific physical contact. In operation, Chi92 should confer selective adherence, increasing contact time between the probiotic and fungal cells. |
| BBa_25RI9TQV | Plasmid_Backbone | pLE-YTK007 - MoClo compatible expression backbone for S. cerevisiae | The plasmid backbone pLE_YTK007 is a shuttle vector for use in both E. coli and S. cerevisiae. It carries a URA3 marker for yeast selection and KanR for bacterial selection. Replication is supported by the 2micron origin in yeast and the ColE1 origin in bacteria. The plasmid includes sfGFP as a fluorescent reporter, along with a ScURA3 terminator for transcriptional termination. Multiple cloning sites with BsaI, BsmBI, and ConS allow efficient Golden Gate assembly. Overall, it enables modular cloning, dual-host propagation, and visual monitoring through GFP expression. |
| BBa_250JF58D | Plasmid_Backbone | ptwist chlor high copy | Twist high copy vector. Works as a backbone in e.coli and B. subtilis in emergencies. Created here to use in composite parts. |
| BBa_25RIMDYU | Coding | YuaB - B. Subtilis Expression pathway with linker sequence | YuaB, a biofilm-associated surface protein of B. subtilis, functions as an effective anchoring motif for bacterial surface display. When fused with a His₆-tag and expressed under a constitutive promoter, YuaB localizes the fusion protein to the bacterial surface, ensuring exposure beyond the peptidoglycan layer. Functionally, YuaB enables the external presentation of heterologous proteins while maintaining host viability and biofilm integrity. More broadly, YuaB provides a stable, surface-exposed, and biofilm-compatible platform that can be adapted for diverse uses in biotechnology, biosensing, and environmental applications. With the linker sequence included this provided a viable pathway to express extracellular proteins. |
| BBa_256RN4XY | Terminator [SO:0000141] | T24 | T24 is an intrinsic (Rho-independent) transcription terminator optimized for use in Bacillus licheniformis. Positioned downstream of coding sequences, it halts RNA polymerase, preventing transcriptional read-through and defining precise 3′ mRNA ends. It is expected to form a stable stem–loop followed by a U-rich sequence, characteristic of intrinsic terminators. With a high termination efficiency of nearly 98%, T24 provides reliable transcriptional insulation, improves mRNA stability, and reduces interference between genetic modules. Its use in expression constructs has been shown to enhance protein yields, reflecting its role in improving transcriptional fidelity and system performance. |
| BBa_25NM1AYT | Coding | SP1 - Non-specific carbohydrate binding protein | SP1 is a synthetic peptide of the same length as the chitinase-derived binding domain (CHI92) designed to serve as an alternate adhesin module in E. coli Nissle. In engineered strains, SP1 is displayed on the cell surface (if a pathway gene is present as well, YuaB for example) and is expected to mediate binding to Candida albicans specifically to its invasive hyphal form, while showing only weak binding to yeast forms or control polymers. As a designed peptide, SP1 does not necessarily carry enzymatic or catalytic activity, but rather functions via its binding affinity to fungal surface components, acting as a synthetic adhesin to tether the probiotic to fungal cells |
| BBa_250JF58D | Primer | Goth010_chi92_mut_REV - Stopcodon mutation primer | Primer designed to mutate in a stop codon after the CHI92 gene |
| BBa_25VJ805I | Primer | Goth009_chi92_mut_FWD - stopcodon mutation primer | Primer designed to mutate in a stop codon after the CHI92 gene |
| BBa_25RZ7TZ2 | Primer | Goth008_Sp1_mut_Rev - Stopcodon mutation primer | Primer designed to mutate in a stop codon after the SP1 gene |
| BBa_25OI51BG | Primer | Goth007_sp1_mut_FWD - Stopcodon mutation primer | Primer designed to mutate in a stop codon after the SP1 gene |
| BBa_25FGFTG0 | Primer | Goth002_chs3_REV - CHS3 amplification primer | Goth002 is an amplification primer for the CHS3 gene. It is meant to amplify the target gene and add scars for MoClo (BsaI and BsmBI). |
| BBa_25RRJ26X | Primer | Goth001_chs3_FWD - CHS3 amplifaction primer | Goth001 is a amplification primer for the CHS3 gene. It is meant to amplify the target gene and add scars for MoClo (BsaI and BsmBI). |
Table of composite parts
Table over composite parts submitted with their corresponding name and description.
| Parts registry ID | Type | Part name | Description |
|---|---|---|---|
| BBa_25WJF999 | Device | Goth03_pLE-YTK007_CHS3 - Expression plasmid for CHS3 in S. cervaise | Goth03 is a yeast expression vector for Saccharomyces cerevisiae, designed to study and enhance chitin biosynthesis in the cell wall. It carries the native CHS3 gene encoding chitin synthase III, the enzyme responsible for chitin polymerization. The construct is derived from the pLE-YTK007 backbone of the Yeast ToolKit (YTK), featuring a yeast origin of replication and modular MoClo assembly sites. URA3 enables auxotrophic selection in yeast, while KanR provides bacterial antibiotic resistance. The original sGFP fragment was replaced with CHS3 using BsaI restriction cloning. This plasmid facilitates controlled CHS3 over expression for functional studies on chitin synthesis, offering a safer S. cerevisiae-based alternative to Candida models. It is suitable for applications in fungal cell wall engineering, metabolic optimization, and synthetic biology. |
| BBa_25K716ND | Device | Goth02_p43_YuaB_Chi92_T24 - Expression plasmid for extracellular binding protein CHI92 | Goth02 is an expression plasmid designed to produce the chitin-binding protein Chitinase 92. Using the Twist high-copy vector as its backbone, it enables high-yield expression in E. coli and B. subtilis. The construct utilizes the YuaB secretion pathway, incorporating a linker to facilitate extracellular expression of CHI92, allowing the protein to interact directly with Candida subspecies. The plasmid carries a CamR resistance marker for selection in bacterial hosts and includes a strong P43 promoter, efficient T24 terminator, and a high-copy ori suitable for robust expression. CHI92 encodes a chitinase capable of binding to chitin, making this plasmid a useful tool for antifungal research, biosensing, and cell-surface binding studies |
| BBa_25H0C19H | Plasmid | Goth01_p43_YuaB_Sp1_T24 - Expression plasmid for extracellular binding protein SP1 | Goth01 is an expression plasmid designed for secretion of the Sp1 binding protein in E. coli and B. subtilis. It uses the Twist high-copy vector backbone for efficient replication and high expression levels. The construct employs the YuaB linker pathway to direct Sp1 to the extracellular environment, enabling binding and interaction with chitin or related fungal cell wall components. The plasmid carries a CamR resistance marker for bacterial selection and includes a strong P43 promoter, a T24 terminator, and a high-efficiency ori optimized for dual-host compatibility. Goth01's aim is to display an extracellular binding protein which binds to fungal cell walls. |