Use of multiple recombination sites with unique specificity in recombinational cloning
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Abstract
The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites with unique specificity. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. Such molecules and/or compounds or combinations of such molecules and/or compounds can also be bound through recombination to various structures or supports according to the invention.
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Citations
155 Claims
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1-142. -142. (Cancelled).
- 143. A composition comprising a solid support having two or more vectors affixed thereon in discrete, defined locations, wherein each of said vectors comprises a DNA insert that is operably linked at each end to at least one promoter.
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144. A composition comprising a solid support having two or more vectors affixed thereon in discrete, defined locations, wherein each of said vectors comprises a first copy and a second copy of a DNA insert each of which is operably linked to a promoter,
wherein transcription of the first copy of said DNA insert results in the generation of a sense transcript, and wherein transcription of the second copy of said DNA insert results in the generation of an antisense transcript.
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145. A composition comprising a solid support having two or more vectors affixed thereon in discrete, defined locations, wherein each of said vectors comprises two copies of a DNA insert that are oriented in opposite orientation relative to each other, and wherein said two insert copies are operably linked to a single promoter.
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146. A composition comprising a solid support having at least a first vector and a second vector affixed thereon in discrete, defined locations, said first vector comprising a copy of a DNA insert operably linked to a promoter, and said second vector comprising a copy of said DNA insert operably linked to a promoter,
wherein transcription of the DNA insert of said first vector results in the generation of a sense transcript, and wherein transcription of the DNA insert of said second vector results in the generation of an antisense transcript.
Specification