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Construction of pET21b::TetR-His6
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DNA containing the coding sequence of TetR (class B, from Tn10) was chemically synthesized (gBlocks dsDNA fragments, IDT). This gBlocks fragment and a NdeI-HindIII-digested pET21b backbone were assembled together using a 2x Gibson master mix (NEB). Gibson assembly was possible due to a 23 bp sequence shared between the NdeI-HindIII-cut pET21b backbone and the gBlocks fragment. These 23 bp regions were incorporated during the synthesis of gBlocks fragments. The resulting plasmids were sequence verified by Sanger sequencing (Eurofins, Germany).
Copyright and License information: ©2020, Jalal et al
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