Adenoviral Vectors

A method for cloning an adenoviral sequence, wherein the adenoviral sequence is a full length adenoviral genome sequence, comprising:

• a) providing a first linear nucleic acid molecule which comprises the full length adenoviral genome sequence;
• b) providing a linearized medium copy plasmid which shares at least two regions of sequence homology with the first linear nucleic acid molecule;
• and
• c) bringing the first linear nucleic acid molecule and the linearized medium copy plasmid into contact in the presence of a 5′ to 3′ exonuclease and an annealing protein such that the first linear nucleic acid molecule and the linearized medium copy plasmid recombine to form a circular plasmid containing the full length adenoviral genome sequence;
• wherein the 5′ to 3′ exonuclease is RecE of SEQ ID NO:
• 1412, or a protein with at least 90% sequence identity to SEQ ID NO:1412, and the annealing protein is RecT,
• and wherein a first region of the at least two regions of sequence homology is a region with sequence homology to only the 5′ ITR of the adenoviral sequence in the first linear nucleic acid molecule, and a second region of the at least two regions of sequence homology is a region with sequence homology to only the 3′ ITR of the adenoviral sequence in the first linear nucleic acid molecule.