Composite

Part:BBa_K2558216

Designed by: Tsinghua 2018   Group: iGEM18_Tsinghua   (2018-10-05)
Revision as of 02:35, 6 October 2018 by Huangtz (Talk | contribs)

CRISPRi Safety Catch device with lux pR-HS promotor

This construct is the “Safety Catch” test system , which is composed of lux pR induced lacI, constantly expressed dCas9 and IPTG inducible gRNA that specifically binds to lux pR promotor on "Neon" system. Without AHL stimulation, gRNA is produced at a basal level and inhibit luxI and GFP (promotor: luxpR-Hypersensitive, or lux pR-HS) transcription. Upon the addition of AHL, lacI on Safety Catch is transcribed (here we need to mention that even though the lacI on the Sately Catch is also controlled by lux pR, the design of gRNA allows us to make it unaffected by CRISPRi, thus it can still be activated by luxR-AHL complex.) and prevents gRNA synthesis, thus relieving inhibition on lux positive feedback system on Neon. We designed this device along with K2558215, which is the same except for the strength of expression and some other parameters.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 310
    Illegal NheI site found at 333
    Illegal NheI site found at 1466
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 5850
    Illegal BamHI site found at 3745
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 79
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4617


[edit]
Categories
Parameters
None