Part:BBa_K2680275

3G tsLambda-cI

Temperature-sensitive Lambda-CI flanked by prefix sticky end C and suffix sticky end D in a 3G-compatible backbone. For more information on 3G hybrid DNA assembly and 3G parts, see the [http://2018.igem.org/Team:William_and_Mary/Part_Collection William and Mary 3G Parts Page].

Lambda-CI is a phage transcriptional repressor that represses lytic pR and pL promoters. Lambda-cI possesses an auto-regulatory function, capable of activating as well as repressing its own transcription when present in sufficient amounts. Temperature-sensitive CI becomes induced at 37C (see induction curve below). We recommend growing it at 29C to avoid unintentional induction.

Figure from Piraner et al. 2016. "OD-normalized fluorescence from the TlpA- and TcI-regulated constructs as a function of induction temperature for a fixed duration of 12 h. N = 4."¹

References

¹Piraner, D. I., Abedi, M. H., Moser, B. A., Lee-gosselin, A., & Shapiro, M. G. (2017). Tunable thermal bioswitches for in vivo control of microbial therapeutics. Nature Chemical Biology, 13(1), 75-80. doi:http://dx.doi.org/10.1038/nchembio.2233

Nandan Kumar Jana, Siddhartha Roy, Bhabatarak Bhattacharyya, Nitai Chandra Mandal; Amino acid changes in the repressor of bacteriophage lambda due to temperature-sensitive mutations in its cI gene and the structure of a highly temperature-sensitive mutant repressor, Protein Engineering, Design and Selection, Volume 12, Issue 3, 1 March 1999, Pages 225–233, https://doi.org/10.1093/protein/12.3.225

Sequence and Features

Assembly Compatibility:

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

  Illegal BsaI site found at 47
  Illegal BsaI.rc site found at 777