Our Part Collection
Our parts collection documents the genetic components designed, characterized, and contributed by our team to the iGEM Parts Registry, forming the foundation of our synthetic biology solution. Together, these parts make up the Assimilatory Nitrate Reduction to Ammonium (ANRA) Pathway, developed to address nitrate reduction in Vibrio natriegens. By registering and sharing this collection, we aim to support not only teams working on nitrate reduction but also contribute to the broader iGEM community. We invite future iGEM teams and researchers to explore and build upon our work, fostering innovation for impactful solutions.
Natronaut’s Parts Collection contains a total of 8 parts: 7 basic parts and 1 composite part. Additionally, 5 parts from other teams were integrated into the design of our composite part. These parts enable the efficient assembly of the ANRA pathway in Vibrio natriegens, catalyzed by three key enzymes.
The parts are categorized into five groups:
Transporter: This enzyme facilitates nitrate uptake from the extracellular environment. It consists of an ABC-type transporter composed of three subunits: nasF, nasE, and nasD.
Nitrate Reductase: Responsible for the reduction of nitrates to nitrites, this enzyme's activity is driven by the molybdoenzyme subunit nasA. The reduction reactions require electron transfer, provided by the NADH-dependent subunit nasC.
Nitrite Reductase: Encoded by nasB, nitrite reductase converts nitrites to ammonium via its heme groups, which accept electrons from donor molecules. A 6x His tag was added for localization in our system.
Plasmid: The plasmid vector, pSEVA261, is a low-copy plasmid with a kanamycin resistance marker and a p15A origin of replication. Its compatibility with V. natriegens was confirmed by Tschirhart et al. (2019) and sourced from the SEVA Plasmid Collection.
Composite Part: Our composite part introduces the ANRA pathway under the control of a single promoter and terminator, optimized for compatibility with our chassis. Between each gene, a native ribosome binding site (RBS) was inserted to regulate expression levels.
These parts provide a robust framework for assimilatory nitrate reduction, offering a valuable resource for future iGEM teams and researchers working in similar domains.