Composite

Part:BBa_K2814020

Designed by: Saksham Sharma   Group: iGEM18_IIT_Delhi   (2018-10-10)


Third Node_ Recombinase based IFFL

Contains the third node for our recombinase based incoherent feedforward loop. It contains sfGFP between the attP and attB sites of TP901-1 and BxbI. Hence, sfGFP gets expressed when recombination takes place once, due to the constitutive P7 promoter in the reverse direction, and stops expressing when the DNA recombines again.

In our case, the third node gets activated when either, BxbI or TP901-1 Integrase is expressed from our first node. The second node gets activated too by the first node, which acts as a repressor on the third node, but starts displaying its function after a time delay. This gives rise to a pulse-like response of our reporter protein which can be easily monitored via fluorescence spectroscopy, thus completing our recombinase-based incoherent feed-forward loop(RIFFL).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 946
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 27
    Illegal BsaI site found at 966
    Illegal SapI.rc site found at 130


[edit]
Categories
Parameters
None