Difference between revisions of "Part:BBa K4083000: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 | ||
Revision as of 12:10, 19 October 2021
pRGPDuo2 plasmid for P. putida
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 3853
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3880
Illegal BamHI site found at 31
Illegal XhoI site found at 2175 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 6480
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Figure 1. Illustration of RPGDuo2 plasmid.
As illustrated in Figure 1, this plasmid contains two multiple cloning sites (MCS 1 and MCS2) with two transcriptional terminators. The MCS1 and MCS2 are regulated by IPTG-inducible Ptac and aTc-inducible PtetR/tetA promoters respectively. Moreover, there are lacI and tetR dependent repression systems that are regulated by Ptac and PtetR/tetA respectively. They were integrated for metabolic engineering purposes. There is a kanamycin resistance gene (KanR). There are two origins of replication to allow replication in E. coli (colE1) and P. putida (pRO1600).
Source
Modified from pRG_Duet for Corynebacterium glutamicum Gauttam et al.
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