Types | DnaRegion
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Roles | Coding
CDS
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Sequences | BBa_K615001_sequence (Version 1)
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Description
This protein is designed for one-hybrid selection systems testing for zinc finger binding. In a strain such as BBa_K615000, where the omega subunit of RNA polymerase has been knocked out, zinc finger binding can be used to control expression of genes necessary for cell survival. Properly bound zinc fingers recruit RNA polymerase to transcribe genes such as His3, allowing the bacteria to grow in incomplete media, while improperly bound zinc fingers will not activate His3 and thus will not survive.
This array is composed of three zinc fingers: Fingers 2 (F2) and 3 (F3) are known from CoDA studies, while F1 is a novel zinc finger designed to bind to the sequence TGG, together creating a zinc finger binding site of 5'-GTG GGA TGG-3'. The part is also designed to allow F1 to be easily replaced by other zinc fingers to allow testing of many different fingers' abilities to bind to the TGG codon.
Notes
Array designed to bind to 5'-GTGGGATGG-3', of which the first six bases represents a known CoDA pair while no binders have yet been found for the last codon in the context of the other fingers.
Source
CoDA sequences from: Jeffry D Sander, Elizabeth J Dahlborg, Mathew J Goodwin et al. Selection-free zinc-finger-nuclease engineering by contextdependent assembly (CoDA). (2011) Nature Methods 8(1): 53-55.