Part:BBa_K2680400

3G spy terminator

Spy terminator ECK120029600 flanked by prefix sticky end D and suffix sticky end E (as well as spacers) 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].

Identified as the strongest natural terminator in supplementary figure 1 of Chen et. al 2013. "Terminator strength (TS) was determined from a simple assay that compares the expression of two fluorescent reporters, one placed before and one after the terminator."¹ According to Explora Biotech, ECK120029600 is a "fairly strong palindromic rho-independent transcriptional terminator of the spheroplast protein y ORF."² ECK120029600 is marked as "recombination-resistant" in a data sheet from by Christopher Voigt's lab at MIT.³

References

¹Chen, Y. J., Liu, P., Nielsen, A. A., Brophy, J. A., Clancy, K., Peterson, T., & Voigt, C. A. (2013). Characterization of 582 natural and synthetic terminators and quantification of their design constraints. Nature methods, 10(7), 659.

²Explora Biotech s.r.l. (2017, January 5). Spy Terminator ECK120029600. Retrieved October 14, 2018, from https://doulix.com/biomodules/GNO9X1/

³Voigt Lab Terminator Plasmids Available from Addgene. (n.d.). Retrieved October 14, 2018, from https://www.addgene.org/synthetic-biology/voigt/terminators/

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 153