RNA

Part:BBa_K1614014:Design

Designed by: Frieda Anna Sorgenfrei   Group: iGEM15_Heidelberg   (2015-09-16)


ATP Aptamer JAWS1 Spinach2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 165
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 165
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 77
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 165
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 165
    Illegal NgoMIV site found at 178
    Illegal NgoMIV site found at 207
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Cloned without restriction enzymes to avoid issues occurring from conserved cut sites. For more details see RFC 110. This part was designed to quantify ATP concentration. To achieve this we exchanged stem two in the Spinach aptamer with an ATP binding aptamer. Now the Spinach can only become fluorescent in presence of ATP. We used this part in our measurement device described on our Wiki. With an appropriate tiration curve the concentration of ATP can directly be calculated of the fluorescent insensitivity.

Source

Synthentic created with JAWS with Spinach II aptamer (Babendure, J.R., Adams, S.R., and Tsien, R.Y. (2003). Aptamers switch on fluorescence of triphenylmethane dyes. Journal of the American Chemical Society 125, 14716-14717.) and ATP aptamer

References