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1 ATGGGGGCAC TGGGGCTGGA GGGCAGGGGT GGGAGGCTCC AGGGGAGGGG TTCCCTCCTG 61 CTAGCTGTGG CAGGAGCCAC TTCTCTGGTG ACCTTGTTGC TGGCGGTGCC TATCACTGTC 121 CTGGCTGTGC TGGCCTTAGT GCCCCAGGAT CAGGGAGGAC TGGGTTTCAG AAGCTGCCAG 181 AGGAGGAGCC AGAAACAGAT CTCAGCCCCG GGCTCCCAGC TGCCCACCTC ATAG
The pGEM-T is 3kb in length, and contains the amplicin resistance gene, conferring selection of the plasmid in E. coli, and the ori site which is the bacterial origin of replication. The plasmid has multiple cloning sites as shown below. The coding sequence was inserted by TA cloning. Many E. coli strains are suitable for the propagation of this vector including JM109, DH5α and TOP10.
The coding sequence can be easily obtained by digesting the vector with proper restriction enzyme(s). The coding sequence can also be amplified by PCR with M13 primers, or primer pair SP6 and T7.