Coding

Part:BBa_K4461002:Design

Designed by: SHIH-HSUN, YANG   Group: iGEM22_NYCU-Taipei   (2022-10-08)


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


References

[1] Tyson K, Busby S, Cole J. Catabolite regulation of two Escherichia coli operons encoding nitrite reductases: role of the Cra protein. Arch Microbiol. 1997 Sep;168(3):240-4. doi: 10.1007/s002030050494. PMID: 9382703. https://pubmed.ncbi.nlm.nih.gov/9382703/

[2]Browning DF, Cole JA, Busby SJ. Suppression of FNR-dependent transcription activation at the Escherichia coli nir promoter by Fis, IHF and H-NS: modulation of transcription initiation by a complex nucleo-protein assembly. Mol Microbiol. 2000 Sep;37(5):1258-69. doi: 10.1046/j.1365-2958.2000.02087.x. PMID: 10972841.

[3]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.