Part:BBa_K4083008

RPGDuo2 plasmid with nadE and rhlBA genes

This plasmid consists of four parts: pRGPDuo2 plasmid; nadE, rhlA, and rhlB coding sequences (https://parts.igem.org/Part:BBa_K4083000, https://parts.igem.org/Part:BBa_K4083004, https://parts.igem.org/Part:BBa_K4083006, https://parts.igem.org/Part:BBa_K4083007). However, for Biobricks purposes, we segmented pRGPDuo plasmid into smaller fragments: TetR/TetA: Part:BBa K2800025 - parts.igem.org; tetR: Part:BBa K4083012 - parts.igem.org; pRO1600: Part:BBa K4083013 - parts.igem.org; LacI reverse: Part:BBa K4083010 - parts.igem.org; Laciq reverse Part:BBa K4083011 - parts.igem.org; Ptac: https://parts.igem.org/Part:BBa_K864400; KanR: https://parts.igem.org/Part:BBa_J31003; ColE1: https://parts.igem.org/Part:BBa_K2560036

By inserting this plasmid, we predicted the effective rhamnolipid production by Pseudomonas putida. Since Pseudomonas putida has all genes required for rhamnolipid production except for rhlAB, the rhlA and rhlb coding sequences inserted in plasmid can allow engineered P. putida to synthesize mono-rhamnolipid by itself. [1]

Reference:

[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

Sequence and Features

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]