Compositions for use in recombinational cloning of nucleic acids
The present invention relates to compositions and methods for use in recombinational cloning of nucleic acid molecules. In particular, the invention relates to nucleic acid molecules encoding one or more recombination sites or portions thereof, to nucleic acid molecules comprising one or more of these recombination site nucleotide sequences and optionally comprising one or more additional physical or functional nucleotide sequences. The invention also relates to vectors comprising nucleic acid molecules of the invention, to host cells comprising vectors or nucleic acid molecules of the invention, to methods of producing polypeptides using nucleic acid molecules of the invention, and to polypeptides encoded by these nucleic acid molecules or produced by methods of the invention. The invention also relates to the use of these compositions in methods for recombinational cloning of nucleic acids, in vitro and in vivo, to provide chimeric DNA molecules that have particular characteristics and/or DNA segments.
1. An isolated nucleic acid molecule comprising a mutated att recombination site including a core region having a nucleotide sequence selected from agcctgctttattatactaagttggcatta (attL5; SEQ ID NO:87) and agcctgcttttttatattaagttggcatta (attL6; SEQ ID NO:88).
2. An isolated nucleic acid molecule comprising a mutated att recombination site including a core region having a nucleotide sequence selected from the group consisting of ggggacaactttgtacaaaaaagttggct (attB1.6; SEQ ID NO: 105), ggggacaactttgtacaagaaagctgggt (attB2.2; SEQ ID NO:97), and ggggacaactttgtacaagaaagttgggt (attB2.10; SEQ ID NO: 107).