Compositions and methods for cloning nucleic acids

A composition comprising a linearized vector comprising a single overhanging dTMP at a 3′ end and a single overhanging dCMP at another 3′ end and a linearized vector comprising a single overhanging dTMP at each 3′ end.

A method of cloning a DNA molecule which is produced by a polymerase chain reaction comprising:

• subjecting a DNA molecule to a polymerase chain reaction producing a nucleic acid product,
• ligating the nucleic acid product in a mixture comprising the nucleic acid product and two or more linear DNA molecules capable of self-replicating when circularized, the linear DNA molecules comprising a single overhanging nucleotide at each 3′ end, wherein a pair of overhanging nucleotides of a linear DNA molecule of the composition are different than a pair of overhanging nucleotides of another linear DNA molecule of the composition.

A kit for cloning of nucleic acids wherein the kit comprises:

• a container which includes a composition comprising two or more linear DNA molecules capable of self-replicating when circularized, the linear DNA molecules comprising a single overhanging nucleotide at each 3′ end, wherein a pair of overhanging nucleotides of a linear DNA molecule of the composition are different than a pair of overhanging nucleotides of another linear DNA molecule of the composition.