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. 2019 Jun 27;38(15):e101649. doi: 10.15252/embj.2019101649

Figure 2. A distal DnaA‐box displays phasing and flexibility.

Figure 2

  1. Systematically shifting a distal rightward‐facing consensus DnaA‐box (CR) by half turns of the DNA double helix. For all constructs, the native 101 bp DNA upstream of DnaA‐box#6 was replaced by a synthetic sequence (“incC art” orange line). Numbering indicates the distance from the middle of DnaA‐box#6 to the middle of the distal DnaA‐box. incC artDnaA‐box#CR22/6/7 (TR711), incC artDnaA‐box#CR28/6/7 (TR710), incC artDnaA‐box#CR33/6/7 (TR326), incC artDnaA‐box#CR39/6/7 (TR325), incC artDnaA‐box#CR44/6/7 (OH70), incC artDnaA‐box#CR49/6/7 (TR324), incC artDnaA‐box#CR55/6/7 (TR186), incC artDnaA‐box#CR60/6/7 (TR328), incC artDnaA‐box#CR65/6/7 (TR329), incC artDnaA‐box#CR70/6/7 (TR330), incC artDnaA‐box#CR75/6/7 (TR184), incC artDnaA‐box#CR80/6/7 (TR709), incC artDnaA‐box#CR86/6/7 (TR708), incC artDnaA‐box#CR91/6/7 (TR707).
  2. Shifting a leftward‐facing consensus DnaA‐box (CL) by half turns of the DNA double helix. incC artDnaA‐box#CL86/6/7 (TR657), incC artDnaA‐box#CL91/6/7 (TR690), incC artDnaA‐box#CL96/6/7 (TR691).
  3. Schematic diagram indicating the replacement of the native DNA sequence between dnaA and DnaA‐box#6 with a larger synthetic sequence (“incC x_art”).
  4. The distal rightward‐facing consensus DnaA‐box (CR) can be shifted several hundred base pairs upstream of its endogenous location. incC x_artDnaA‐box#CR44/6/7 (TR210), incC x_artDnaA‐box#6/7 (TR209), incC x_artDnaA‐box#CR132/6/7 (TR203), incC x_artDnaA‐box#CR297/6/7 (TR206), incC x_artDnaA‐box#CR462/6/7 (TR208).