Part:BBa_K4461003:Design

glpABC promoter

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 cloned a larger fragment that include the promoter, then use another pair of primers to specify clone of the promoter. Therefore we can avoid the length of the unintended bands being too similar to distinct easily. The sequence we cloned has to include all transcriptional factors for the promoter. We use BBa_B0034 as the RBS of the promoter, so we didn't clone the RBS from the genomic DNA.

Source

The glpABC promoter is cloned from E.coli K-12 MG1655 genomic DNA.

References

[1]Smith A, Kaczmar A, Bamford RA, Smith C, Frustaci S, Kovacs-Simon A, O'Neill P, Moore K, Paszkiewicz K, Titball RW, Pagliara S. The Culture Environment Influences Both Gene Regulation and Phenotypic Heterogeneity in Escherichia coli. Front Microbiol. 2018 Aug 15;9:1739. doi: 10.3389/fmicb.2018.01739. PMID: 30158905; PMCID: PMC6104134.

[2]Poladyan, A., Avagyan, A., Vassilian, A. et al. Oxidative and Reductive Routes of Glycerol and Glucose Fermentation by Escherichia coli Batch Cultures and Their Regulation by Oxidizing and Reducing Reagents at Different pHs. Curr Microbiol 66, 49–55 (2013).

[3]Khademian, M., and Imlay, J. A. (2017) Escherichia coli cytochrome c peroxidase is a respiratory oxidase that enables the use of hydrogen peroxide as a terminal electron acceptor. Proc Natl Acad Sci U S A 114: E6922–E6931.

[4]Alternative respiratory pathways of Escherichia coli: energetics and transcriptional regulation in response to electron acceptors Biochim. Biophys. Acta Bioenerg., 1320 (1997), pp. 217-234