title: AssemblX: a user-friendly toolkit for rapid and reliable multi-gene assemblies.
abstract: The assembly of large DNA constructs coding for entire pathways poses a major challenge in the field of synthetic biology. Here, we present AssemblX, a novel, user-friendly and highly efficient multi-gene assembly strategy. The software-assisted AssemblX process allows even unexperienced users to rapidly design, build and test DNA constructs with currently up to 25 functional units, from 75 or more subunits. At the gene level, AssemblX uses scar-free, overlap-based and sequence-independent methods, allowing the unrestricted design of transcriptional units without laborious parts domestication. The assembly into multi-gene modules is enabled via a standardized, highly efficient, polymerase chain reaction-free and virtually sequence-independent scheme, which relies on rare cutting restriction enzymes and optimized adapter sequences. Selection and marker switching strategies render the whole process reliable, rapid and very effective. The assembly product can be easily transferred to any desired expression host, making AssemblX useful for researchers from various fields.
authors.affiliation: University of Potsdam, Cell2Fab Research Unit, Karl-Liebknecht-Strasse 24-25, 14476 Potsdam, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Hochrein
authors.affiliation: University of Potsdam, Cell2Fab Research Unit, Karl-Liebknecht-Strasse 24-25, 14476 Potsdam, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Machens
authors.affiliation: Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam, Germany.
authors.forename: Juergen
authors.initials: J
authors.lastname: Gremmels
authors.affiliation: University of Potsdam, Cell2Fab Research Unit, Karl-Liebknecht-Strasse 24-25, 14476 Potsdam, Germany.
authors.forename: Karina
authors.initials: K
authors.lastname: Schulz
authors.affiliation: University of Potsdam, Cell2Fab Research Unit, Karl-Liebknecht-Strasse 24-25, 14476 Potsdam, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Messerschmidt
authors.affiliation: Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam, Germany.|University of Potsdam, Department of Molecular Biology, Karl-Liebknecht-Strasse 24-25, 14476 Potsdam, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Mueller-Roeber
mesh_terms: D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods*; D020451:Contig Mapping / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005819:Genetic Markers; D016678:Genome; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D012984:Software
chemical_list: D005819:Genetic Markers; D011994:Recombinant Proteins; D004247:DNA; D004262:DNA Restriction Enzymes