Regulatory
ts-feedbac

Part:BBa_K3285002:Design

Designed by: Sayantan Datta   Group: iGEM19_IISER-Pune-India   (2019-10-16)


Temperature sensitive negative feedback loop using cI-lam promoter


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 AgeI site found at 630
    Illegal AgeI site found at 742
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The RBS sites were too small to be extracted from Biobrick and stitched using PCR and hence were introduced with standard scar sequences through primers.


Source

The part was made by using the following:- a) Promoter - cI-lam from Biobrick BBa_R0051 present in distribution plate-6 (5K); b) mRFP gene from Biobrick BBa_E1010 present in distribution plate-3 (12M); c) cI-ts2 (Biobrick BBa_K3285001) which was synthesised from TwistBio; d) Double Terminator from Biobrick BBa_B0015 present indistribution plate-3 (4E). RBS were introduced using primers during PCR amplification and stitching.

References