Difference between revisions of "Part:BBa K2558215"

 
Line 1: Line 1:
 +
__NOTOC__
 +
<partinfo>BBa_K2558215 short</partinfo>
  
 +
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: lux pR) 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 K2558216, which is the same except for the strength of expression and some other parameters.
 +
 +
<!-- Add more about the biology of this part here
 +
===Usage and Biology===
 +
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K2558215 SequenceAndFeatures</partinfo>
 +
 +
 +
<!-- Uncomment this to enable Functional Parameter display
 +
===Functional Parameters===
 +
<partinfo>BBa_K2558215 parameters</partinfo>
 +
<!-- -->

Revision as of 02:35, 6 October 2018

CRISPRi Safety Catch device with lux pR 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: lux pR) 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 K2558216, 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 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1163
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 5547
    Illegal BamHI site found at 3442
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 5777
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4314