New Genes for TALENs

For cloning of TALENs we have developed a binary vector which harbors both DNAs on one plasmid. Cloning  of the  TALE repeat domain can be done either in the SacI-BbvCI or into the AscI-HindIII restriction sites. The present vector contains the Ubi-maize promoter for transformation of monocots. But the TALENs cassette can be simply transfered with SfiI restriction to any other binary containing these restriction sites. Both  backbones of the TALENs construct were codon optimized for high expression in monocots and dicots. They are identical in amino acid sequence. But to  avoid problems due to repeated sequences, both backbones are not identical in the sequence of the nucleic acid. Both 35s promoters can be easily exchanged. We recommend small promoters, since cloning can be difficult, when the complete construct reaches 20 kb.

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