Difference between revisions of "Part:BBa K5348005"

 
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pL-RBS2
 
  
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===Usage and Biology===
 
  
 
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===Functional Parameters===
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    <h2>BBa_K5348005 (pL-RBS2)</h2>
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    <h3>Construction Design</h3>
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        To reduce the leaky expression of the light-inducible induction system (BBa_K3447133, hereafter referred to as the pL), we reduced the intensity of the RBS linked to the target gene in this element by 108-fold (RBS2). The composition of this element is shown below.
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        <img src="https://static.igem.wiki/teams/5348/bba-k5348005/figure-1.jpg" alt="Figure 1. Schematic diagram of the pL-RBS2">
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        <div class="caption">Figure 1. Schematic diagram of the pL-RBS2</div>
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    <h3>Engineering Principle</h3>
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        Under dark condition, histidine kinase (YF1) phosphorylates FixJ (response regulator of histidine kinase), which activates PFixK2 (the target gene for transcription upon FixJ activation), driving the expression of the cI gene (λ phage repressor), which represses the transcription of its cognate promoter, PR (the cognate promoter of cI), and downstream genes cannot be expressed. Under blue light, the cI gene cannot be expressed, PR can be transcribed normally, and downstream genes can be expressed [1].
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    <h3>Reference</h3>
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    <p>[1] H, Mays RL, Hoffman SM, Avalos JL. Optogenetic Control of Microbial Consortia Populations for Chemical Production. ACS Synth Biol. 2021 Aug 20;10(8):2015-2029.</p>
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Revision as of 09:34, 30 September 2024


pL-RBS2


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1874
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 587
    Illegal NgoMIV site found at 659
    Illegal NgoMIV site found at 749
    Illegal NgoMIV site found at 767
    Illegal NgoMIV site found at 1259
    Illegal NgoMIV site found at 1552
    Illegal NgoMIV site found at 1646
    Illegal AgeI site found at 301
    Illegal AgeI site found at 1427
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1316
    Illegal BsaI.rc site found at 200


<!DOCTYPE html> BBa_K5348005 (pL-RBS2)

BBa_K5348005 (pL-RBS2)

Construction Design

To reduce the leaky expression of the light-inducible induction system (BBa_K3447133, hereafter referred to as the pL), we reduced the intensity of the RBS linked to the target gene in this element by 108-fold (RBS2). The composition of this element is shown below.

Figure 1. Schematic diagram of the pL-RBS2
Figure 1. Schematic diagram of the pL-RBS2

Engineering Principle

Under dark condition, histidine kinase (YF1) phosphorylates FixJ (response regulator of histidine kinase), which activates PFixK2 (the target gene for transcription upon FixJ activation), driving the expression of the cI gene (λ phage repressor), which represses the transcription of its cognate promoter, PR (the cognate promoter of cI), and downstream genes cannot be expressed. Under blue light, the cI gene cannot be expressed, PR can be transcribed normally, and downstream genes can be expressed [1].

Reference

[1] H, Mays RL, Hoffman SM, Avalos JL. Optogenetic Control of Microbial Consortia Populations for Chemical Production. ACS Synth Biol. 2021 Aug 20;10(8):2015-2029.