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Golden Gate Assembly

Golden Gate Assembly. 1 ul (50 ng ) Receiving plasmid 1 ul (1:1 molar ratio) annealed oligos 1 ul 10X Ligase buffer 0.5 ul BsaI HF 0.5 ul T4 DNA Ligase 6.0 ul dH2O 10 ul Total. 20 cycles 37° C for 1 minute 16° C for 1 minute 37° C for 15 minutes. pTac Promoter.

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Golden Gate Assembly

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  1. Golden Gate Assembly 1 ul (50 ng) Receiving plasmid 1 ul (1:1 molar ratio) annealed oligos 1 ul 10X Ligase buffer 0.5 ulBsaI HF 0.5 ul T4 DNA Ligase 6.0 ul dH2O 10 ul Total 20 cycles 37° C for 1 minute 16° C for 1 minute 37° C for 15 minutes

  2. pTac Promoter • Hybrid promoter constructed from pTrp and pLac promoters in E. coli -35 -10 +1 J119137 BsaI BsaI GFP RFP DeBoer HA, Comstock LJ, Vasser M (1982) PNAS 80, 21-25.

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