Part:BBa_K5477051
Team UCopenhagen 2024: Parts Collection of MilkClear
Introduction
Our part collection includes 48 total parts: 24 basic parts, 16 composite parts, and 8 devices, with 45 of which are new. This collection is coherent, as all the parts were developed with the specific goal to address contaminants in breast milk. It is also the first collection tackling this purpose.
The parts can be categorized into three main categories: 1.) Parts for detecting contaminants in breast milk. 2.) Parts for detoxifying contaminants. 3.) Devices integrating these functions.
Instead of function, we have chosen complexity to group our parts on this page, as multiple parts reappear in each category, due to the interlinked nature of our collection. All parts have been tested in either the wet-lab or the dry-lab, which demonstrates their function. We also cloned additional constructs that are not part of this collection.
By contributing a comprehensive collection of receptor, reporter, and detoxification modules, we provide valuable resources that expand current biosensing capabilities and empower future iGEM teams to develop more sophisticated, multi-functional systems.
Devices
The Devices Table outlines the different biosensor devices developed for the detection and detoxification of various environmental contaminants. Each device incorporates specific receptor modules and reporter modules, tailored to detect particular compounds like polycyclic aromatic hydrocarbons (PAHs), dioxins, polychlorinated biphenyls (PCBs), and bisphenol A (BPA). For instance, devices BBa_K5477041 and BBa_K5477042 are biosensors designed to detect PAHs, dioxins, or dioxin-like PCBs, while devices BBa_K5477043 to BBa_K5477045 are specialized for BPA detection. Additionally, BBa_K5477046 introduces the SUPERMOM device, which integrates both detection and detoxification modules to simultaneously identify and neutralize BPA in breast milk. The final device, BBa_K5477047, is a detoxification device targeting PAHs, dioxins, and PCBs through the use of CYP1A1 and UGT enzymes, which metabolize these contaminants into safer, water-soluble compounds. This array of devices demonstrates the versatility of the biosensor platform in addressing diverse environmental and health-related issues.
| Part Name | Part | Type | Function |
|---|---|---|---|
| BBa_K5477041 | BBa_K5477023: pSTE12-AhR - receptor module, BBa_K5477025: pRET2-ARNT receptor module, BBa_K5477026: pRET2-NCOA - receptor module, BBa_K5477030: XRE-pMEL1-NanoLuc reporter module | Device | Biosensor device for detection of PAHs, dioxin or dioxin-like PCBs |
| BBa_K5477042 | BBa_K5477024: pRAD27-AhR - receptor module, BBa_K5477025: pRET2-ARNT receptor module, BBa_K5477026: pRET2-NCOA - receptor module, BBa_K5477030: XRE-pMEL1-NanoLuc reporter module | Device | Biosensor device for detection of PAHs, dioxin or dioxin-like PCBs |
| BBa_K5477043 | BBa_K5477027: pRET2-LexA-ERα(LBD) - receptor module, BBa_K5477031: Lex6Op-pLEU2-NanoLuc reporter module | Device | Biosensor device for detection of BPA |
| BBa_K5477044 | BBa_K5477028: pPOP6-LexA-ERα(LBD) - receptor module, BBa_K5477031: Lex6Op-pLEU2-NanoLuc reporter module | Device | Biosensor device for detection of BPA |
| BBa_K5477045 | BBa_K5477029: pRET2-LexA-mERα(LBD) - receptor module, BBa_K5477031: Lex6Op-pLEU2-NanoLuc reporter module | Device | Biosensor device for detection of BPA |
| BBa_K5477046 | BBa_K5477029: pRET2-LexA-mERα(LBD) - receptor module, BBa_K5477031: Lex6Op-pLEU2-NanoLuc reporter module, BBa_K5477040:UDPD-pPDC1-Lex6Op-pENO1-UGT2B15 | Device | SUPERMOM: Dual-Function Biosensor for BPA Detection and Detoxification |
| BBa_K5477047 | BBa_K5477037: CYP1A1-pGAL1/10-POR detox module, BBa_K5477036: UDPD-pGAL1/10-UGT1A1 detox module | Device | Detoxification device against dioxin and PCBs |
| BBa_K5477048 | pRET2-LexA-mERα(LBD) - receptor module, BBa_K5477031: Lex6Op-pLEU2-NanoLuc reporter module, BBa_K5477039: CYP3A4-MYC-pPDC1-Lex6Op-pENO1-POR | Device | Testing of the SUPERMOM Concept |
Composites
The Composites Table showcases the various composite parts that make up the functional modules of each biosensor device. Each composite part consists of a combination of promoters, receptors, or detoxification modules designed for specific detection or detoxification tasks. For example, BBa_K5477023 and BBa_K5477024 feature receptor modules tailored to detect PAHs, dioxins, and dioxin-like PCBs, while BBa_K5477027 and BBa_K5477028 are optimized for BPA detection through engineered estrogen receptor modules. Additionally, the composite parts BBa_K5477035 to BBa_K5477039 focus on detoxification, containing enzymes such as UGT2B15, UGT1A1, and CYP1A1 that metabolize harmful chemicals into less toxic or more easily excretable forms. These composite parts serve as the building blocks of the devices, providing the necessary functionality to detect and detoxify environmental contaminants.
| Part Name | Part | Type | Length (bp) | |
|---|---|---|---|---|
| BBa_K5477023 | pSTE12-AhR - receptor module | Composite | Promoter and Receptor for detection of PAHs, dioxin or dioxin-like PCBs | 3117 |
| BBa_K5477024 | pRAD27-AhR - receptor module | Composite | Promoter and Receptor for detection of PAHs, dioxin or dioxin-like PCBs | 1900 |
| BBa_K5477025 | pRET2-ARNT receptor module | Composite | Promoter and nuclear translocator detection of PAHs, dioxin or dioxin-like PCBs | 2155 |
| BBa_K5477026 | pRET2-NCOA - receptor module | Composite | Promoter and nuclear receptor co-activator for the detection of PAHs, dioxin or dioxin-like PCBs | 5032 |
| BBa_K5477027 | pRET2-LexA-ERα(LBD) - receptor module | Composite | Promoter and Receptor for detection of BPA | 2389 |
| BBa_K5477028 | pPOP6-LexA-ERα(LBD) - receptor module | Composite | Promoter and Receptor for detection of BPA | 2389 |
| BBa_K5477029 | pRET2-LexA-mERα(LBD) - receptor module | Composite | Promoter and Receptor for detection of BPA | 2389 |
| BBa_K5477030 | XRE-pMEL1-NanoLuc - reporter module | Composite | A xenobiotic response element, a promoter, and a luciferase reporter gene for the detection signal against PCBs, dioxins, and dioxin-like compounds | 868 |
| BBa_K5477031 | Lex6Op-pLEU2-NanoLuc reporter module | Composite | DNA-binding domain, promoter, and luciferase reporter gene for the detection signal against BPA | 880 |
| BBa_K5477035 | UDPD-pGAL1/10-UGT2B15 | Composite | Detoxification module against BPA | 3724 |
| BBa_K5477036 | UDPD-pGAL1/10-UGT1A1 | Composite | Detoxification module against PAHs, dioxin and PCBs | 3726 |
| BBa_K5477037 | CYP1A1-pGAL1/10-POR | Composite | Detoxification module against PAHs, dioxin and PCBs | 4312 |
| BBa_K5477038 | CYP3A4-pGAL1/10-POR | Composite | Detoxification module against a wide array of contaminants | 4250 |
| BBa_K5477039 | CYP3A4-MYC-pGAL1/10-POR | Composite | Detoxification module against a wide array of contaminants | 4286 |
| BBa_K5477040 | UDPD-pPDC1-Lex6Op-pENO1-UGT2B15 | Composite | Detoxification module for the detoxification of BPA | 3631 |
| BBa_K5477032 | CYP3A4-MYC-pPDC1-Lex6Op-pENO1-POR | Composite | Detoxification module for BPA | 4193 |
Parts
The Parts Table lists individual genetic components, including promoters, coding sequences (CDS), and regulatory elements, that make up the biosensor and detoxification systems. Promoters such as pRET2 and pSTE12 are used to drive constitutive or inducible expression of the receptor and detoxification modules, ensuring that the biosensors are responsive under the desired conditions. The table also includes various coding sequences for receptors like AhR, ARNT, and LexA-ERα, which are central to detecting endocrine disruptors like BPA and other toxic chemicals. Detoxification enzymes such as CYP1A1, UGT2B15, and CYP3A4 are also featured, responsible for metabolizing and neutralizing harmful substances. These individual parts allow the biosensors to be highly modular, enabling customization for different contaminants while maintaining efficiency and compatibility with standard assembly methods.
| Part Name | Part | Type | Length (bp) |
|---|---|---|---|
| BBa_K5477000 | pRET2 - Medium strong constitutive promoter in Saccharomyces cerevisiae | Regulatory (Promoter) | 700 |
| BBa_K5477001 | pSTE12 - Constitutive promoter in Saccharomyces cerevisiae | Regulatory (Promoter) | 557 |
| BBa_K5477002 | pRAD27 - Weak constitutive promoter in Saccharomyces cerevisiae | Regulatory (Promoter) | 694 |
| BBa_K5477003 | pMEL1 - Constitutive core promoter in Saccharomyces cerevisiae | Regulatory (Promoter) | 256 |
| BBa_K5477004 | pLEU2 - Constitutive core promoter in Saccharomyces cerevisiae | Regulatory (Promoter) | 130 |
| BBa_K5477005 | pGAL1/10 - Bidirectional inducible promoters from Saccharomyces cerevisiae | Regulatory (Promoter) | 664 |
| BBa_K5477006 | pPDC1-Lex6Op-pENO1 - Bidirectional synthetic promoters with Lex6Op | Regulatory (Promoter) | 571 |
| BBa_K5477007 | pPOP6 - Weak constitutive promoter in Saccharomyces cerevisiae | Regulatory (Promoter) | 700 |
| BBa_K5477008 | XRE - Xenobiotic Response Element | Regulatory Binding Element | 82 |
| BBa_K5477009 | Lex6Op - six tandem copies of LexA operator | Regulatory Binding Element | 220 |
| BBa_K3793004 | AhR - Aryl hydrocarbon Receptor | CDS - receptor | 2547 |
| BBa_K3793005 | ARNT - Aryl hydrocarbon Nuclear Transporter | CDS - receptor | 2370 |
| BBa_K1680009 | NanoLuc - NanoLuciferase reporter gene | CDS - Reporter gene | 516 |
| BBa_K5477012 | NCOA - Nuclear receptor coactivator | CDS - receptor | 4326 |
| BBa_K5477013 | LexA-ERα chimeric activator | CDS - receptor | 1683 |
| BBa_K5477014 | LexA-mERα chimeric activator | CDS - receptor | 1683 |
| BBa_K5477015 | LexA-ERRγ chimeric activator | CDS - receptor | 1602 |
| BBa_K5477016 | UGT2B15 | CDS - enzyme | 1593 |
| BBa_K5477017 | UGT1A1 | CDS - enzyme | 1602 |
| BBa_K5477018 | UDPD | CDS - enzyme | 1446 |
| BBa_K5477019 | CYP1A1 | CDS - receptor | 1575 |
| BBa_K5477020 | CYP3A4 | CDS - enzyme | 1512 |
| BBa_K5477021 | CYP3A4-MYC | CDS - enzyme | 1548 |
| BBa_K5477022 | POR | CDS - enzyme | 2061 |
| None |