Part:BBa_K2066111

pLacO1 sfGFP + LacI (weak)

This part places both pLacO1 driving sfGFP expression (K2066014) and consititively expressed LacI onto the same plasmid. This reduces the metabolic stress on transformed cells and helps limit the number of plasmids used by the circuit control toolbox. Placing all circuit control parts on a single plasmid will allow a workable number of plasmids to be used for the input circuit to the circuit control toolbox circuits.

Source: The sfGFP flourescent reporter and pLacO1 design is inspired by C. Lou, B. Stanton, Y.-J. Chen, B. Munsky, C. A. Voigt, Ribozyme-based insulator parts buffer synthetic circuits from genetic context. Nat. Biotechnol. 30, 1137 (2012). doi:10.1038/nbt.2401 pmid:23034349. The UNS sequences at the ends of the insert are derived from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860. A huge thanks to all the researchers involved in its original creation! The LacI (BBa_C0012) sequence is from the iGem registry.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1087
  Illegal NheI site found at 1110
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 224