Synthetic perturbation of gene expression is central to our ability to reliably uncover genotype-phenotype relationships in microbes. Here, I present a novel gene transcription activation strategy that uses the Vibrio cholerae CRISPR-Associated Transposon (CAST) system to selectively insert promoter elements upstream of genes of interest. Through this strategy, I show robust activation of both recombinant and endogenous genes across the Escherichia coli chromosome. I then demonstrate precise tuning of expression levels by exchanging the promoter elements being inserted. Finally, I demonstrate that CAST activation can be used to synthetically induce ampicillin-resistant phenotypes in E. coli.