IGEM:IMPERIAL/2009/Assays Protocols/Cloning Strategy M1&T

Module 1 Cloning Strategy

• Ligate PAH/ Cellulase gene sequence (G1) from GeneArt with the arabinose-inducible promoter from the iGEM starter kit.
• Test for successful ligation using Cellulose/ PAH assay as defined here
• If successful, remove terminators from G1 gene sequence using suitable restriction enzymes.
• Ligate sequence to LacI (BBa_C0012) from the iGEM starter kit.
• Test for expression of LacI gene using LacI repressible sequence coupled with reporter gene. BBa_I13601 is such a reporter - coupled with CFP.

Preliminary Schematic

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