The invention provides cells and methods of circularizing linear DNA
molecules. The cell is an isolated Escherichia coli cell which
transiently expresses the Cre recombinase protein from an integrated Cre
recombinase gene, and which is at least transiently repressed for RecBCD
activity. The cells are used in a method of circularizing a linear DNA
molecule comprising at least two loxP sites. The DNA molecule is
introduced into the cells, and the linear DNA molecule is joined at said
loxp sites.