Patent Document ID: 8053191
Application ID: 11897939
Patent Flag: 1

Claim One:
1. A method for assembling nucleic acid segments, the method comprising providing a first and second population of nucleic acids, each population having at least a first, second, third and fourth restriction sites, wherein each population comprises at its 5′ end a 5′ activation sequence located between the first and second restriction sites and at its 3′ end a 3′ activation sequence located between the third and fourth restriction sites; generating a first population of nucleic acid segments having the 5′ activation sequence but lacking the 3′ activation sequence by digesting the first population of nucleic acids using a first set of restriction enzymes that cleave the nucleic acids at the first and third restriction sites; generating a second population of nucleic acid segments having the 3′ activation sequence but lacking the 5′ activation sequence by digesting the second population of nucleic acids using a second set of restriction enzymes that cleave the nucleic acids at the second and fourth restriction sites; combining the first and second populations of nucleic acid segments with a first linearized nucleic acid vector that is digested with one or more restriction enzymes that produce restriction site overhangs that are complementary to the overhangs generated by the first and fourth restriction enzymes on the first and second populations of nucleic acid segments, wherein the first linearized nucleic acid vector comprises (i) at its 3′ end a coding sequence of a first marker gene 5′ of the first restriction site, and (ii) at its 5′ end a coding sequence of a second marker gene 3′ of the fourth restriction site, and isolating ligated first nucleic acid vectors that express the first and the second marker genes, wherein expression of the first and the second marker genes is indicative of correct assembly of the first and second populations of nucleic acid segments and wherein expression of the first marker gene is regulated by the first activation sequence and expression of the second marker gene is regulated by the second activation sequence.