https://pages.htgaa.org/2026a/sergio-cuiza/labs/lab-week-6--gibson-assembly/index.htmlCompletion status: This lab was completed virtually (in silico primer design, virtual PCR, Gibson assembly simulation, and sequence analysis). The physical wet lab (PCR thermocycling, DpnI digest, DNA purification, Gibson assembly, transformation of E. coli, and plate incubation) was not performed. All results below are theoretical, based on the published paper (Liljeruhm et al., 2018) and the provided protocol. Overview & Objective In this two‑day lab, we change the chromophore of the purple Acropora millepora chromoprotein (amilCP) to orange, pink, and blue mutants by PCR‑based mutagenesis and Gibson assembly. The amilCP gene is carried on the mUAV plasmid (Addgene). We amplify two fragments – a backbone (origin, chloramphenicol resistance, promoter, RBS) and an insert (chromophore region + terminator) – with overlapping ends. The insert forward primer contains the desired mutation(s). After DpnI digestion (to remove template plasmid), we purify the fragments, assemble them via Gibson, and transform into chemically competent E. coli. Only cells with the correctly assembled plasmid survive on chloramphenicol and express coloured chromoproteins.Hugoen