Method For Constructing And Modifying Large Dna Molecules

Abstract

This Invention Provides A Method For Combining Overlapping Dna Molecules Comprising: (A) Providing First And Second Dna Fragments, The First Having A Region Homologous To A Region In The Second; (B) Tagging The First Dna Fragment With A Selectable Marker; (C) Cloning The First Dna Sequence Into A Retrieval Vector To Form A Dna-Vector Complex; (D) Linearizing The Dna-Vector Complex; And (E) Inserting The First Dna Fragment From The Dna-Vector Complex Into The Second Dna Fragment Using Homologous Recombination To Form A Combined Dna Molecule; And (F) Removing The Selectable Marker, Thereby Generating A Combined Dna Molecule. The Invention Further Provides A Vector For Retrieving And Inserting A Selected Dna Molecule Into A Target Dna Molecule.