DNA

Part:BBa_K5443028

Designed by: Nick Coleman, Carmen Hawthorne   Group: iGEM24_MacquarieUni   (2024-10-01)

This composite part consists of five genes (ECH-Ppu, FCS-Ppu, COMT-ara, C3H-Sam5 and TAL-Sam8), each with a strong RBS attached, cloned into an expression vector (pUS250) to make the construct pMQ3A. The RBS used are all slightly different variants of B0034, designed to be of predicted strength 10,000 (arbitrary units) using the DeNovoDNA website RBS calculator. The genes are organised into a two-gene cluster (ECH-Ppu + FCS-Ppu) and three-gene cluster (COMT-ara, C3H-Sam5, TAL-Sam8), with each cluster under the control of its own cumate-inducible promoter. The two cumate promoter sequences are intended to be functionally identical but the non-essential sequence elements have been modified in the second promoter to minimise plasmid mutations due to homologous recombination.

Figure 1. Structure of composite part K5443028: Insert DNA in Vanillin biosynthetic plasmid pMQ3A

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