Week 2 HW: DNA Design

The name of my repository: Ira-Dhawan-hydrogel-project2026 The name of my folder: IraDhawanHydrogel

What molecule are you expressing?

I am expressing a protein that forms the hydrogel mesh, CsgA. This is the major curli amyloid subunit, which self-assembles outside the cell into a water-retaining hydrogel that can serve as the basis for a wound-care material. This directly impelents my week 1 idea.

What is the sequence? Give the protein or RNA sequence, or the specific gene, you will encode, and say where it comes from (a database accession such as UniProt or NCBI, a known part, or de novo). Cite it.

Sequence: E. Coli CsgA protein, UniProt accession P0AEK3. I will use the full 151 amino acid sequence which includes the natural signal peptide. Source: UniProt database, known natural part, not de novo.

What host or chassis? The living system from HW1, for example E. coli, S. cerevisiae, CHO or another mammalian line, or a cell-free system. Your host drives almost every part choice that follows.

E. coli- same microbial system I proposed in Week 1. It’s fast, well-characterized, and naturally produces curli.

Where does the product need to go? Cytoplasm, secreted, membrane, or a compartment? This decides whether you need a signal peptide or targeting sequence.

Sectreted/extracellular. Hydrogel must form outside the cell,so I need a signal peptide. CsgA already has its native Sec signal peptide (amino acids 1-20). So if I use PelB, I will note it. So targeting - secreted to periplasm/extracellular via Sec pathway.