Patent ID: 7981670

Claim:
A kit comprising: two expression vectors selected from the group consisting of (1) an expression vector comprising a promoter operably linked to a DNA construct that comprises a translation initiation codon, a multiple cloning site downstream of the initiation codon, a mutation hotspot sequence downstream of the initiation codon, and a bacterial colony reporter gene downstream of the mutation hotspot sequence, wherein when a DNA sequence of interest is introduced into the DNA construct at the multiple cloning site, a protein that comprises the amino acid sequence encoded by the reporter gene and maintains the function of the protein product of the reporter gene can be produced from the initiation codon in a bacterial cell and a loss-of-function mutation in the mutation hotspot sequence or the DNA sequence of interest destroys the function of the reporter gene, (2) an expression vector comprising a promoter operably linked to a DNA construct that comprises a translation initiation codon, a mutation hotspot sequence downstream of the initiation codon, and a bacterial colony reporter gene downstream of the mutation hotspot sequence, wherein the translation initiation codon, the mutation hotspot sequence, and the reporter gene are arranged in a way so that a protein that comprises the amino acid sequence encoded by the reporter gene and maintains the function of the protein product of the reporter gene can be produced from the initiation codon in a bacterial cell and a loss-of-function mutation in the mutation hotspot sequence destroys the function of the reporter gene, and (3) an expression vector comprising a promoter operably linked to a DNA construct that comprises a translation initiation codon, a multiple cloning site downstream of the initiation codon, and a bacterial colony reporter gene downstream of the initiation codon, wherein when a DNA sequence of interest or a mutation hotspot sequence is introduced into the DNA construct at the multiple cloning site a protein that comprises the amino acid sequence encoded by the reporter gene and maintains the function of the protein product of the reporter gene can be produced from the initiation codon in a bacterial cell and a loss-of-function mutation in the DNA sequence of interest or the mutation hotspot sequence destroys the function of the reporter gene, wherein the two expression vectors are (1) and (2), expression vectors (1) and (3), or expression vectors (2) and (3); and an instruction manual on using two of said vectors for assessing mutability of a DNA sequence of interest.