Part:BBa_K4461004:Design
ansB promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 117
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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 ansB promoter is cloned from E.coli K-12 MG1655 genomic DNA.
References
[1]Jennings MP, Beacham IR. Co-dependent positive regulation of the ansB promoter of Escherichia coli by CRP and the FNR protein: a molecular analysis. Mol Microbiol. 1993 Jul;9(1):155-64. doi: 10.1111/j.1365-2958.1993.tb01677.x. PMID: 8412660.
[2]Robbins JW Jr, Taylor KB. Optimization of Escherichia coli growth by controlled addition of glucose. Biotechnol Bioeng. 1989 Dec 5;34(10):1289-94. doi: 10.1002/bit.260341007. PMID: 18588069.
[3]Bradley MD, Beach MB, de Koning APJ, Pratt TS, Osuna R. Effects of Fis on Escherichia coli gene expression during different growth stages. Microbiology (Reading). 2007 Sep;153(Pt 9):2922-2940. doi: 10.1099/mic.0.2007/008565-0. PMID: 17768236.
[4]Oliveira, E.M.M., Carvajal, E. & Bon, E.P.S. L-Asparaginase II of Saccharomyces cerevisiae . Appl Biochem Biotechnol 77, 311–316 (1999).
[5]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.