Composite

Part:BBa_K2066040:Design

Designed by: Kalen Clifton, Christine Gao, Andrew Halleran, Ethan Jones, Likhitha Kolla, Joseph Maniaci, John Marken, John Mitchell, Callan Monette, Adam Reiss   Group: iGEM16_William_and_Mary   (2016-10-14)


J23100 + RiboJ Promoter Characterization Part with 2x Broccoli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 47
    Illegal NheI site found at 70
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

UNS standard chosen for ease of cloning with standard primers.

Source

This part was created using gibson assembly from existing parts.

The UNS2, UNS3, and UNS6 sequences are taken from Torella et al. 2013 (“Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly”). RiboJ sequence is from Lou et. al 2012 ("Ribozyme-based insulator parts buffer synthetic circuits from genetic context"). The spacer sB is taken from Pothoulakis et al. 2013 (“The Spinach RNA Aptamer as a Characterization Tool for Synthetic Biology”). The broccoli sequence is from https://www.addgene.org/66843/. The mCherry sequence is taken from K2066029 with the start codon and extra stop codon added.

References