Difference between revisions of "Part:BBa K3320006"
 
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<partinfo>BBa_K3320006 short</partinfo>
 
<partinfo>BBa_K3320006 short</partinfo>
  
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This is a multi-input AND-gate for biosensor tumor microenvironment.
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This is a multi-input AND-gate for tumor microenvironment.
  
 
<!-- Add more about the biology of this part here <!-- -->
 
<!-- Add more about the biology of this part here <!-- -->
 
===Usage and Biology===
 
===Usage and Biology===
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Our AND Gate has repressor-promoter relation, basically using two promoters to control the signal (repressor) production that represses our protein of interest.
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<p>Our AND Gate has a repressor-promoter relation, basically using two promoters to control the signal (repressor) production that represses our protein of interest. It is composed of two major units: 1) Lactate repressor unit, and 2) Hypoxia repressor unit.</p> 
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<p>The lactate repressor unit is composed of the lactate promoter, inducible in the presence of lactate, producing the PhlF repressor protein. This protein represses the PhlF promoter, which is responsible for producing the Tet repressor protein, that regulates the expression of GFPmut3 by Tet promoter. </p>
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<p>The hypoxia repressor unit is composed of the fdhF promoter, inducible in low concentrations of oxygen, expressing the Srp repressor protein, which represses the Srp promoter producing TetR that regulates the expression of GFPmut3 by Tet promoter.</p>
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<p>Therefore, our system is in a constant state of repression, except in the presence of lactate and hypoxia, which induces the production of GFPmut3. These are physiological characteristics of the tumor microenvironment. </p>
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===Assembly===
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<p>This part was cloned on pUC19 vector, totaling 8004 bp.</p>
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[[File:BBa_K3320006_Circuit.png|550px|]]
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<p> Figure 1. BBa_K3320006 plasmidial map.</p>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3320006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3320006 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 14:57, 21 October 2019


Multi-input AND-gate

This is a multi-input AND-gate for tumor microenvironment.

Usage and Biology

Our AND Gate has a repressor-promoter relation, basically using two promoters to control the signal (repressor) production that represses our protein of interest. It is composed of two major units: 1) Lactate repressor unit, and 2) Hypoxia repressor unit.

The lactate repressor unit is composed of the lactate promoter, inducible in the presence of lactate, producing the PhlF repressor protein. This protein represses the PhlF promoter, which is responsible for producing the Tet repressor protein, that regulates the expression of GFPmut3 by Tet promoter.

The hypoxia repressor unit is composed of the fdhF promoter, inducible in low concentrations of oxygen, expressing the Srp repressor protein, which represses the Srp promoter producing TetR that regulates the expression of GFPmut3 by Tet promoter.

Therefore, our system is in a constant state of repression, except in the presence of lactate and hypoxia, which induces the production of GFPmut3. These are physiological characteristics of the tumor microenvironment.


Assembly

This part was cloned on pUC19 vector, totaling 8004 bp.

BBa K3320006 Circuit.png

Figure 1. BBa_K3320006 plasmidial map.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 78
    Illegal NheI site found at 101
    Illegal NheI site found at 808
    Illegal NheI site found at 831
    Illegal NheI site found at 4308
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1415
    Illegal AgeI site found at 4057
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3284