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Fig. 3 | Microbial Cell Factories

Fig. 3

From: Direct pathway cloning of the sodorifen biosynthetic gene cluster and recombinant generation of its product in E. coli

Fig. 3

DiPaC of the sod BGC into pET28b-ptetO::sod_gfpv2. a PCR amplification of the 4.6 kb sodAD fragment with Q5 polymerase and primers containing homology sequences for SLIC into the backbone, b sodA–D fragment after gel purification, c PCR amplification to construct a linear 6.6 kb backbone of pET28b-ptetO::gfpv2, d positive screening PCR of five clones after transformation of the SLIC-joined pET28b-ptetO::sod_gfpv2 plasmid and e control restriction digest of two clones with NdeI or EcoRV in comparison to the predicted (Snapgene) pattern

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