CloneFast: A simple plasmid design and construction guide for labs venturing into synthetic biology.

Traditional plasmid assembly often involves expensive or restrictive enzymatic processes. To overcome these limitations, we introduce the CloneFast guide, a streamlined approach for efficient plasmid construction. Initially, this guide advises users through bioinformatic plasmid and oligonucleotide design in silico. Subsequently, we outline practical experimental steps, including the introduction of phosphorothioate modifications via reusable modified oligonucleotides (CompetePCR), generation of precise sticky ends through a rapid iodine-mediated cleavage, and seamless fragment assembly into plasmids. CloneFast enables the completion of plasmid assembly within 3 days, offering versatility and accessibility to a wide range of research groups.