Method and kits for preparing multicomponent nucleic acid constructs

A kit for the production of nucleic acid multicomponent constructs, comprising a package containing nucleic acid components, each component comprising a double stranded nucleic acid molecule having at least one single stranded 5′ or 3′ terminal sequence the terminal sequence having sufficient complementarity to either a terminal sequence in a separate nucleic acid component or to a sequence in a linking nucleic acid molecule so to allow for specific annealing and linkage of the components in a predetermined order.

A kit for the production of nucleic acid multicomponent constructs, comprising at least 3 different nucleic acid components appropriately phosphorylated for ligation, the kit further comprising a ligase enzyme.

A kit for the production of vectors, comprising nucleic acid components, each component comprising a double stranded nucleic acid molecule having at least one single stranded 5′ or 3′ terminal sequence, the terminal sequence having sufficient complementarity to either a terminal sequence in a separate nucleic acid component or to a sequence in a linking nucleic acid molecule so as to allow for specific annealing and linkage of the components in a predetermined order, wherein the nucleic acid components encode:

• i) an origin of replication, and ii) a selectable marker

A method of linking nucleic acid components in a predetermined order to produce a nucleic acid multicomponent construct, comprising:

• (a) providing the nucleic acid components and one or more linking nucleic acid molecules to be assembled into the construct, each nucleic acid component comprising a double stranded nucleic acid molecule having at least one single stranded 5′ or 3′ terminal sequence, the terminal sequence having sufficient complementarity to a sequence in a linking nucleic acid molecule so as to allow for specific annealing of complementary sequences and linkage of the components in a predetermined order;
• (b) incubating the nucleic acid components under conditions which allow for the specific annealing and linkage of the nucleic acid components to thereby produce the nucleic acid multicomponent construct.