Team:William and Mary/Basic Part

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Basic Part

Bba_K0026550 is a decoy binding array containing 85 TetO repeat sequences, and was derived from an addgene plasmid from Finney-Manchester et al. (2013) (“Harnessing mutagenic homologous recombination for targeted mutagenesis in vivo by TaGTEAM). This part can be used to shift the transfer function of an aTC inducible promoter to the left (Figure 1). See decoy binding array page for more information.

This part was assembled using restriction cloning with EcoRI and SpeI, as accurately performing PCR on the sequence is close to impossible due to the large stretches of homologous sequences.

Figure 1: Population level FACs data comparing the relative fluorescence of a pTet sfGFP and TetR reporter with and without a tetO binding array. While the data is noisy, it is clear that the inflection point of the circuit with the binding array has shifted to the left as expected. Additionally, both circuits experienced a decrease in fluorescence at higher aTC concentrations, this is likely due to the toxicity of aTC in high concentrations. (See binding array page for more details)

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