Difference between revisions of "Part:BBa K4083008:Design"
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===References=== | ===References=== | ||
| − | [1] Gauttam, R., Mukhopadhyay, A., & Singer, S. W. (2020). Construction of a novel dual-inducible duet-expression system for gene (over)expression in Pseudomonas putida. Plasmid, 110. https://doi.org/10.1016/j.plasmid.2020.102514 | + | [1]Tiso, T., Sabelhaus, P., Behrens, B., Wittgens, A., Rosenau, F., Hayen, H., & Blank, L. M. (2016b). Creating metabolic demand as an engineering strategy in Pseudomonas putida – Rhamnolipid synthesis as an example. Metabolic Engineering Communications, 3, 234–244. https://doi.org/10.1016/j.meteno.2016.08.002 |
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| + | [2] Gauttam, R., Mukhopadhyay, A., & Singer, S. W. (2020). Construction of a novel dual-inducible duet-expression system for gene (over)expression in Pseudomonas putida. Plasmid, 110. https://doi.org/10.1016/j.plasmid.2020.102514 | ||
Latest revision as of 12:33, 19 October 2021
RPGDuo2 plasmid with nadE and rhlBA genes
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 5565
Illegal BamHI site found at 6125
Illegal XhoI site found at 1757
Illegal XhoI site found at 2024
Illegal XhoI site found at 6301 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1949
Illegal NgoMIV site found at 2019
Illegal NgoMIV site found at 2240
Illegal NgoMIV site found at 4400 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
https://static.igem.org/mediawiki/parts/4/49/BBa_K4083008-pRGPDuo2-Edited_sequence.pdf
We planned to insert rhlA and rhlB genes to MCS1 and nadE gene to MCS2 of pRGPDuo2 plasmid. Thus, the rhlA and rhlB genes are controlled by the IPTG-inducible Ptac promoter, while the nadE gene is controlled by aTc-inducible PtetR/teA promoter. Those promoters are regulated by lacI and tetR repression systems
Source
This plasmid was constructed by our team by using the pRGPDuo2 plasmid provided by Gauttam, R.
References
[1]Tiso, T., Sabelhaus, P., Behrens, B., Wittgens, A., Rosenau, F., Hayen, H., & Blank, L. M. (2016b). Creating metabolic demand as an engineering strategy in Pseudomonas putida – Rhamnolipid synthesis as an example. Metabolic Engineering Communications, 3, 234–244. https://doi.org/10.1016/j.meteno.2016.08.002
[2] Gauttam, R., Mukhopadhyay, A., & Singer, S. W. (2020). Construction of a novel dual-inducible duet-expression system for gene (over)expression in Pseudomonas putida. Plasmid, 110. https://doi.org/10.1016/j.plasmid.2020.102514