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The most basic form of our project

The most basic form of our project. Mefp-3. Tyrosinase. Mefp-5. Cu chaperone?. Mgfp-5. The most basic form of our project: operon-style. Regulatory signal. Mefp-3. Mefp-5. Mgfp-5. Cu chaperone. Tyrosinase. For B. subtilis chassis.

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The most basic form of our project

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  1. The most basic form of our project Mefp-3 Tyrosinase Mefp-5 Cu chaperone? Mgfp-5

  2. The most basic form of our project: operon-style Regulatory signal Mefp-3 Mefp-5 Mgfp-5 Cu chaperone Tyrosinase

  3. For B. subtilis chassis • Add 30-aa tag at N-terminus of all protein coding sequences • Use B. subtilis-specific promoters, terminators (?), ribosome binding sites (?) • Any other signaling components (e.g. quorum-sensing machinery) will have to work in B. subtilis

  4. Existing parts of interest: • BBa_J51000 – ComP B. subtilis quorum sensor coding sequence; nonstandard prefix and suffix sites • BBa_J51001 – ComA, the effector of ComP; nonstandard prefix and suffix sites • BBa_J51002 – ComQX operon—produces the secreted quorum sensing signal • BBa_J33206 – ars promoter and arsR gene plus E. coli lacZ • BBa_I742103 – Gram negative/positive shuttle vector chloramphenicol, neomycin & gentamicin resistance markers • BBa_I742123: pTG262-BB: biobrick vector derived from pTG262 • BBa_I746665 - B. subtilis strong promoter – inducible by IPTG (presumably only if you introduce LacI) • BBa_I746666 – B. subtilis strong promoter driving GFP

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