Patent ID: 7160702

Claim:
A method for transferring a nucleic acid fragment of interest from Parent Molecule 1 to Parent Molecule 2 wherein Parent Molecule 1 comprises the nucleic acid fragment of interest and Parent Molecule 2 comprises at least one functional element that is able to influence the transcription, translation or replication of the nucleic acid fragment of interest, or for transferring a nucleic acid fragment of interest from Parent Molecule 2 to Parent Molecule 1 wherein Parent Molecule 2 comprises the nucleic acid fragment of interest and Parent Molecule 1 comprises at least one functional element that is able to influence the transcription, translation or replication of the nucleic acid fragment of interest, wherein Parent Molecule 1 may be linear or circular, and Parent Molecule 2 may be linear or circular; wherein Parent Molecule 1 comprises a first recombination site and Parent Molecule 2 comprises a second recombination site; wherein Parent Molecule 1 if linear comprises a first joinable end (JE1), and if circular comprises a first region (CS1) that can be converted into a first joinable end; and wherein Parent Molecule 2 if linear comprises a second joinable (JE2) end and if circular comprises a second region (CS2) that can be converted into a second joinable end; the method comprising: (1) mixing Parent Molecule 1 and Parent Molecule 2 in vitro; (2) forming an intermediate molecule, by either (a) site-specific recombination between the first and second recombination sites, or by (b)(i) joining the first and second joinable ends, or (ii) first converting the region or regions that can be converted into a joinable end or joinable ends, and then joining the joinable ends, the intermediate molecule comprising the nucleic acid molecule of interest and the at least one functional element; and (3) processing the intermediate by either (a) joining the first and the second joinable ends, or (b) by site-specific recombination between the first and second recombination sites, to form a circular Product Vector comprising the nucleic acid molecule of interest and the at least one functional element.