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Standard BioBrick plasmid backbone

Goal: To persuade a bacterium to produce a variety of small peptides and secrete them into the medium. Peptide secretor cassette. X. E. P. S. antibiotic resistance. origin. Standard BioBrick plasmid backbone. (http://partsregistry.org/Plasmids). His. M. Cassette Structure.

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Standard BioBrick plasmid backbone

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  1. Goal: To persuade a bacterium to produce a variety of small peptides and secrete them into the medium

  2. Peptide secretor cassette X E P S antibiotic resistance origin Standard BioBrickplasmid backbone (http://partsregistry.org/Plasmids)

  3. His M Cassette Structure Secretory signal GFP Peptide RBS

  4. His M Standard Promoter • e.g. J23100 – Strong, constitutive promoter Secretory signal GFP Peptide RBS

  5. His M Standard RBS • e. g. B0034 – star 1 and W Secretory signal GFP Peptide RBS

  6. His M Synthesize Secretory Signal • e. g. lipA or sacB – identified from registry • Possibly need to synthesize • Leave as an exercise for the student! Secretory signal GFP Peptide RBS

  7. His M Some random gene • Need a signaller… • … and a handle • Allow student contributions for alternatives Secretory signal GFP Peptide RBS

  8. His M The ‘slot’ in the cassette • Need: • 1 cleavage site • 1 target peptide • 1 purification handle Secretory signal Peptide GFP RBS

  9. His M What goes in the player? • The polymer engineer is thinking about, e.g. extracellular matrix proto-molecules and self-assembling elements • The biologist is thinking about using this as a recruiting tool… Secretory signal Peptide GFP RBS

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