Part:BBa_K1620001:Design

Promoter element Zasp

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

We have identified two sequences of this promoter (one in sense strand and another in the antisense strand). In order to turn this sequence more feasible we used a hybrid final sequence. The final sequence was believed to promote for both sites. A previously report of Gilston et al. (2014) have shown that another sequence could be promoter this operon in the direct sense (Figure 1).

Figure 1: Sequence aligment of the promoters Zasp and that determined for znuABC by Gilston et al. (2014).

Source

The promoter element was designed through bioinformatic analysis of 500bp frame upstream genes of operon ZnuABC from E. coli K12 MG1655 genome, as observed in Figure 2. Then, it was synthesized by IDT through G-block and further assembled by 3A assembly method.

Figure 2: Genome of E. coli K-12 MG1655 showing the operon genes znuABC and the region of where the Zasp promoter was obtained.

References

Gilston BA, Wang S, Marcus MD, Canalizo-Hernández MA, Swindell EP, Xue Y, et al. 2014. Structural and Mechanistic Basis of Zinc Regulation Across the E. coli Zur Regulon. PLoS Biol 12(11): e1001987. doi:10.1371/journal.pbio.1001987