Coding

Part:BBa_K2333007:Design

Designed by: Ethan M Jones   Group: iGEM17_William_and_Mary   (2017-06-29)

Super Folder GFP (sfGFP) stop codons removed


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 SapI.rc site found at 13


Design Notes

sfGFP has a much faster folding time (~6 min) compared to wild type GFP.[2] This makes it preferable as a reporter for many circuits since your fluorescence readout will more accurately indicate the given cellular concentration of reporter. Greater photobleaching than wild type GFP has been reported in the literature (~20%).[3] However, for most experiments this is not a significant concern.

Source

>The sequence for this sfGFP 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." Sequence was modified to remove any restriction enzymes and then synthesized by IDT


References

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

[2] http://book.bionumbers.org/what-is-the-maturation-time-for-fluorescent-proteins/

[3] Pedelacq, J. D.; Cabantous, S.; Tran, T.; Terwilliger, T. C.; Waldo, G. S. Nat. Biotechnol. 2006, 24, 1170−1170.