BioSample

Protocols: The reaction (20 µL) was performed with 0.08 pmol of attB or attP plasmid DNA library, 0.08 pmol of supercoiled plasmid containing WT attB or attP, 7.2 pmol of mv4Int and 40 µg of crude-extract from E. coli BL21(DE3) heated at 95°C for 10 min, and incubated 1h30 at 42°C. . PCR products were purified using the QIAquick PCR purification kit (Qiagen, Germany) The different random DNA libraries were constructed as follow: randomized oligonucleotides (109-bp in length for attB libraries; 184-bp in length for core-attP libraries, Supplementary Table S3B) were obtained by chemical synthesis (Integrated DNA Technologies, USA). PCR was used to create double-stranded DNA using specific primers (attBLib-F / attBLib-R pair for attB and attPLib-F / attPLib-R pair for attP, see accompanying paper. Each PCR-product population was separately cloned by Gibson assembly, either into pCC1FOS™ plasmid (Epicentre, USA) for attB libraries, or plasmid pMET359 (see accompanying paper) for attP libraries. Clones were propagated in E. coli strain EPI300 (Epicentre, USA) under chloramphenicol selection for attB libraries, or in E. coli strain MET961 (see accompanying paper) under carbenicillin selection for attP libraries.