Reporter

Part:BBa_K3380150:Design

Designed by: Alexandru Popov   Group: iGEM20_Edinburgh   (2020-10-08)


iSpinach fluorescent RNA aptamer


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
    COMPATIBLE WITH RFC[1000]


Design Notes

We ordered the synthesized sequence. Originally ordered sequence: forward (5’TACT agacgcgactacggtgagggtcgggtccagtagcttcggctactgttgagtagagtgtgggctccgtagtcgcgtctc 3’ ) and (5’CATTgagacgcgactacggagcccacactctactcaacagtagccgaagctactggacccgaccctcaccgtagtcgcgtct 3’) with TACT and CATT respectively being a linker, to facilitate further manipulations (swapping of the part or attaching to different promoters).

The forward and reverse sequences were phosphorylated, annealed and ligated to other parts using T4 DNA ligase.


The iSpinach RNA aptamer was flanked by the F30 tRNA scaffold BBa_K3380101 and BBa_K3380102 for increased fluorescence and stability.

Source

iSpinach is an artificial RNA aptamer that was obtained using the SELEXapproach. It is based on the Spinach fluorescent RNA aptamer.

References

Autour, A., Westhof, E. and Ryckelynck, M., 2016. iSpinach: a fluorogenic RNA aptamer optimized for in vitro applications. Nucleic acids research, 44(6), pp.2491-2500.