Overview
Using one of the notable promoters found at the end of the screenings and titrations, our team theorized an example plasmid that could be used as a biosensor using these promoters, and this plasmid was entered into iGEM’s part catalog under “Plasmids” for future iGEM teams to use and to track our work. The promoters we used in our project are from “A comprehensive library of fluorescent transcriptional reporters for Escherichia coli” [1], and corresponding sequences were found through EcoCyc [2]. In our plasmid, every part except for the promoter and reporter sfGFP was sourced from our team’s last year parts that are currently in iGEM’s parts catalog: RepA (origin of replication) is BBa_K4651005, pSC101 (origin of replication) is BBa_K4651006, rrnB1 (terminator) is BBa_K4651009, bla (antibiotic resistance marker for carbenicillin) is BBa_K4651003, and bla promoter (promoter for bla resistance marker) is BBa_K4651004. The sequence for the reporter sfGFP was found through the NCBI [3]. To create the plasmid map, PlasMapper 3.0 [4] was used.
Parts Table
| Name | Type | Description | Designers | Length |
|---|---|---|---|---|
| pIGEM1, or BBa_K5240000 | Plasmid | Example biosensor using promoter gadB, which could be used to detect butanoyl-homoserine lactone, with a resistance against carbenicillin and reporter sfGFP | Jason Gao | 5306 |
Plasmid/Part Sequence
References:
[1] Zaslaver, A., Bren, A., Ronen, M., Itzkovitz, S., Kikoin, I., Shavit, S., Liebermeister, W., Surette, M. G., &
Alon, U. (2006). A comprehensive library of fluorescent transcriptional reporters for Escherichia coli.
Nature Methods, 3(8), 623–628. DOI: 10.1038/nmeth895
[2] Keseler, I. M., Gama-Castro, S., Mackie, A., Billington, R., Bonavides-Martínez, C., Caspi, R., Kothari, A.,
Krummenacker, M., Midford, P. E., Muñiz-Rascado, L., Ong, W. K., Paley, S., Santos-Zavaleta, A., Subhraveti, P.,
Tierrafría, V. H., Wolfe, A. J., Collado-Vides, J., Paulsen, I. T., & Karp, P. D. (2021). The EcoCyc Database in
2021. Frontiers in Microbiology, 12, 711077. DOI: 10.3389/fmicb.2021.711077
[3] National Center for Biotechnology Information (NCBI) [Internet]. Bethesda (MD): National Library of Medicine
(US), National Center for Biotechnology Information; [1988]. Available from: www.ncbi.nlm.nih.gov
[4] Wishart, D. S., Ren, L., Leong-Sit, J., et al. (2023). PlasMapper 3.0 - A Web Server for Generating, Editing,
Annotating and Visualizing Publication Quality Plasmid Maps. Nucleic Acids Research. DOI:
10.1093/NAR/GKAD276