Cloning and/or sequencing vector

Abstract

A cloning and/or sequencing vector enables recombinant clones to be selected directly. The vector encodes a fusion protein which includes a protein poison.

1 . A method of cloning a DNA fragment in a prokaryotic cell, said method comprising:
 providing a cloning vector comprising at least one promoter nucleotide sequence and at least one nucleotide sequence comprising at least one cloning site localized at the level of a nucleotide sequence for a gene encoding CcdB, said CcdB being cytotoxic;   inserting said DNA fragment into said at least one cloning site in said cloning vector such that said DNA fragment interferes with the cytotoxic activity of said CcdB;   transferring the cloning vector with the DNA fragment into the prokaryotic cell; and   growing the prokaryotic cell to produce multiple copies of the DNA fragment.   
     
     
         2 . The method of  claim 1 , wherein said at least one nucleotide sequence comprises a plurality of cloning sites. 
     
     
         3 . The method of  claim 1 , wherein said at least one cloning site is a unique cloning site. 
     
     
         4 . The method of  claim 1 , wherein said at least one cloning site is an artificially created cloning site. 
     
     
         5 . The method of  claim 1 , wherein said at least one promoter nucleotide sequence comprises a repressible promoter. 
     
     
         6 . The method of  claim 5 , wherein said repressible promoter is a Lac operon promoter. 
     
     
         7 . The method of  claim 1 , wherein said cloning vector is a recombinant virus or recombinant plasmid. 
     
     
         8 . The method of  claim 7 , wherein said recombinant plasmid comprises a pUC origin of replication. 
     
     
         9 . The method of  claim 8 , wherein said pUC origin of replication is the origin of replication from pUC18 or pUC19.