Composite

Part:BBa_K4083008:Design

Designed by: Arsen Orazbek   Group: iGEM21_NU_Kazakhstan   (2021-10-18)
Revision as of 12:23, 19 October 2021 by Arsenorazbek (Talk | contribs) (→‎References)


RPGDuo2 plasmid with nadE and rhlBA genes


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE 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
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE 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] 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