Difference between revisions of "Part:BBa K2120402"
 
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<partinfo>BBa_K2120402 short</partinfo>
 
<partinfo>BBa_K2120402 short</partinfo>
  
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This is one of our application circuit.
 
  
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This circuit consists of two parts:the inhibitor device and killer device ,which have already been made into basic parts of biobricks.
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'''<Description>'''
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<br>This is one of our application circuits.
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<br>This circuit contains two devices: inhibitor device and killer device.
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<br>Inhibitor device: We select mediun weak constitutive promoter J23116 and strong RBS B0034 to express inhibitor gene ''tetR''.
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<br>Killer device: Toxin protein, MazF, can cleavage ACA sequence of free mRNA specifically. It cause bacteria can’t translate successfully and dead finally. So we constructed it with induced promoter pTet and strong RBS B0034 to control the expression level of toxin protein.
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<br>We reversed the direction of inhibitor device, in order to reduce the influence between two devices.
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<br>Meanwhile, we also constructed more circuits which have different combine with promoter, RBS and inhibitor. More details see in our Wiki([http://2016.igem.org/Team:BIT-China/Parts Parts Submission])
  
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We choose three different promoters (BBa_J23109,BBa_J23116,BBa_J23106) whose reported strengths are respectively low, medium and strong from constitutive promoter family to express the inhibitor tetR and cI ,and then we construct three plasmids with different copy number containing this circuit, in order to find out the proper promoter which meets the threshold that trigger the killer system in E.coli. According to our results about the lethal effect of killer device , [https://parts.igem.org/wiki/index.php?title=Part:BBa_K212008 MazF] ,so we replace the killer gene with mRFP reporter gene in order to figure out the regulation relationship between two devices.
 
  
 
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Latest revision as of 12:26, 23 October 2016


B0015+tetR+B0034+J23116+ptet+B0034+mazF+B0015


<Description>
This is one of our application circuits.


This circuit contains two devices: inhibitor device and killer device.
Inhibitor device: We select mediun weak constitutive promoter J23116 and strong RBS B0034 to express inhibitor gene tetR.
Killer device: Toxin protein, MazF, can cleavage ACA sequence of free mRNA specifically. It cause bacteria can’t translate successfully and dead finally. So we constructed it with induced promoter pTet and strong RBS B0034 to control the expression level of toxin protein.
We reversed the direction of inhibitor device, in order to reduce the influence between two devices.
Meanwhile, we also constructed more circuits which have different combine with promoter, RBS and inhibitor. More details see in our Wiki([http://2016.igem.org/Team:BIT-China/Parts Parts Submission])


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 851
    Illegal NheI site found at 874
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]