https://pages.htgaa.org/2026b/mariana-catalina-pemberthy-mantilla/week-02-hw-dna-design/index.htmlPart 0: Set Up in Kernel Repository & Folder Location: My project repository and folder in the HTBAA 2026 Workspace is located at: HTBAA26_WilliamandMary / Mariana Catalina Pemberthy Mantilla. Part 1: Define Your Expression Target What molecule are you expressing? I am expressing a set of three metabolic enzymes (PhaA, PhaB and PhaC) that form the core biosynthetic pathway to manufacture polyhydroxyalkanoate (PHA) bioplastic for absorbent fibers and waterproof backing films in zero-waste menstrual pads. What is the sequence? Organism Source: Cupriavidus necator (formerly Ralstonia eutropha) H16. Database Accession Numbers: phaC (PHA synthase): UniProt P23608 (NCBI Gene ID: 1080383). phaA (3-ketothiolase): UniProt P14611 (NCBI Gene ID: 1080381). phaB (Acetoacetyl-CoA reductase): UniProt P14697 (NCBI Gene ID: 1080382). What host or chassis? Escherichia coli BL21(DE3). In Cell-Free Metabolic Engineering (CFME), this host chassis is grown in cultures to overexpress the target pathway enzymes; then, cells are lysed to harvest an active enzyme extract for in vitro biomanufacturing. Where does the product need to go? In Vitro Reaction Solution (Cytoplasmic Extract). The target enzymes accumulate inside the host cell cytoplasm during extract preparation and operate freely in the open cell-free reaction vessel. No secretion signal tag or membrane-targeting sequence is required. Part 2: Design Your Genetic Parts Every genetic construct requires specific control elements assembled around the coding sequences. Below is the parts table corresponding to the five individual parts created in Kernel for high-level expression in E. coli BL21(DE3):Hugoen