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. 2017 Nov 1;12(11):e0186568. doi: 10.1371/journal.pone.0186568

Fig 3. TA-GC cloning.

Fig 3

A: BccI recognizes the sequence CCATC at the cloning site of pET-BccI and cuts out at the recognition site as indicated. B: after digestion with BccI. C: the protein-coding gene (starting with an ATG codon and always having a 3΄ glycine-coding GGC codon) after PCR amplification, and D: after treatment with T4 DNA polymerase in the presence of dATP and dGTP. The 5΄-Α and G overhangs of the protein-coding gene created after incubation with the T4 DNA polymerase are complementary to the 5΄-T and C overhangs of the pET-BccI vector after digestion with BccI.