Measurement

Part:BBa_K2333420:Design

Designed by: Sejal Dhawan   Group: iGEM17_William_and_Mary   (2017-10-27)


UNS pTet sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 996
    Illegal NheI site found at 1019
  • 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 133


Design Notes

This part was designed with the aTc inducible promoter pTet for sfGFP reporter combined with the pTet repressor tetR under the control of the medium-weak strength constitutive promoter J23105.The sfGFP reporters have been codon-optimized for E. coli and feature a double stop codon for enhanced efficiency.

Source

UNS sequences are 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.

The sequence for this sfGFP (BBa_K2333007) was obtained and modified from the sequence in Supplement section V of Lou et al. "2015 Ribozyme-based insulator parts buffer synthetic circuits from genetic context." The sequence was modified to remove any restriction enzymes and then synthesized by IDT.

References

[1] Torella JP, Boehm CR, Lienert F, Chen J-H, Way JC, Silver PA. Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic Acids Research. 2013;42(1):681–689.

[2] Lou, C., Stanton, B., Chen, Y.-J., Munsky, B., & Voigt, C. A. (2012). Ribozyme-based insulator parts buffer synthetic circuits from genetic context. Nature Biotechnology, 30(11), 1137–1142. http://doi.org/10.1038/nbt.2401