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ftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/013/SRR29236413/SRR29236413.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/070/SRR29236570/SRR29236570_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/091/SRR29236791/SRR29236791_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/051/SRR29236551/SRR29236551.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/059/SRR29236659/SRR29236659.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/036/SRR29236536/SRR29236536.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR292/074/SRR29236674/SRR29236674.fastq.gzprimaryOK200GenomicsBiomedical Engineering, Boston Universityhttps://www.ebi.ac.uk/ena/browser/view/PRJNA1118281xref:PubMed:40335485The DNA binding of most Escherichia coli Transcription Factors (TFs) has not been comprehensively mapped, and few have models that can quantitatively predict binding affinity. We report the global mapping of in vivo DNA binding for 139 E. coli TFs using ChIP-Seq. We used these data to train BoltzNet, a novel neural network that predicts TF binding energy from DNA sequence. BoltzNet mirrors a quantitative biophysical model and provides directly interpretable predictions genome-wide at nucleotide resolution. We used BoltzNet to quantitatively design novel binding sites, which we validated with biophysical experiments on purified protein. We have generated models for 125 TFs that provide insight into global features of TF binding, including clustering of sites, the role of accessory bases, the relevance of weak sites, and the background affinity of the genome. Our paper provides new paradigms for studying TF-DNA binding and for the development of biophysically motivated neural networks. Overall design: Chromatin immunoprecipitation DNA-sequencing (ChIP-Seq) of 139 TFs from Escherichia coli K-12 MG1655 under native and inducible expression modes, plus in vitro ChIP-Seq of one TF, and Library-ChIP of 3 TFs.ENAfalsePredictive Biophysical Neural Network Modeling of a Compendium of in vivo Transcription Factor DNA Binding Profiles for Escherichia coliPredictive Biophysical Neural Network Modeling of a Compendium of in vivo Transcription Factor DNA Binding Profiles for Escherichia coli2025-05-182025-03-22PRJNA1118281GSE26869840335485