High-throughput assembly of genetic elements

A set of sequence-verified nucleic acid elements for combinatorial construction of genetic elements, said set comprising two or more nucleic acid elements assembled from a plurality of nucleic acid parts, comprising one or more categories of nucleic acid parts, wherein the nucleic acid elements comprise:

• at least two sequences selected from the plurality of nucleic acid parts;
• (ii) a 3′ flanking nucleic acid sequence and a 5′ flanking nucleic acid sequence, which together are capable of functioning to order assembly of higher-order nucleic acid constructs, to facilitate retrieval, and to identify the nucleic acid element,
• wherein the 3′ and 5′ flanking sequences comprise one or more nucleic acid barcodes that are unique to each sequence-verified nucleic acid element, which together are capable of functioning to facilitate retrieval and to identify the nucleic acid element, and
• wherein the 3′ and 5′ flanking sequences comprise one or more restriction sites that are capable of being cleaved prior to assembly in order to produce sticky ends to facilitate ordered assembly of higher order nucleic acid elements;
• and
• (iii) at least one DNA hash,
• wherein each DNA hash comprises a unique nucleic acid barcode capable of functioning to identify either the nucleic acid element or a nucleic acid part in the nucleic acid element,
• wherein the barcode of each DNA hash is flanked by primer binding sites capable of being amplified to produce an amplicon capable of forming a concatemer with an amplicon from another DNA hash in another nucleic acid element,
• wherein the primer binding sites are different for nucleic acid elements in the set of sequence-verified nucleic acid elements that are for each position within a combinatorial construction of genetic elements;
• and
• wherein each DNA hash is located internally to the cleavage sites for the one or more restriction sites in (ii), whereby the hash cannot be removed by restriction enzyme digestion at the one or more restriction sites.