Part:BBa_M36855:Design
Recombinase-based heat-inducible sensor
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1545
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1765
Illegal NheI site found at 1788
Illegal PstI site found at 1545 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1545
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1545
Illegal NgoMIV site found at 1712
Illegal AgeI site found at 1599 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1732
Illegal BsaI.rc site found at 1815
Design Notes
During the design of this part, we carefully selected an irreversible integrase so that the sequence encoding the constitutive promoter could not be flipped back to the "wrong" direction (in which it doesn't trigger transcription of the downstream actuator), and once triggered would theoretically remain "on" for the rest of the organism's lifespan. We used a strong RBS to ensure robust expression of the integrase under heat conditions, and purposefully selected a well-characterized heat inducible promoter from the Parts Registry. A strong constitutive promoter was used between the recombinase sites so that detection of a "flipping" event would be as easy as possible.
Source
This part is an amalgamation of existing Registry parts, attB and attP sites, and the Bxb1 integrase from the cited PNAS paper, which we've also made into a part and submitted to the registry.
References
Bonnet J, Subsoontorn P, Endy D. 2014. Rewritable Digital Data Storage In Live Cells Via Engineered Control Of Recombination Directionality. PNAS. 109(23): 8884-8889.