Difference between revisions of "Part:BBa K3278001"
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In this part, we use the T7 promoter to substitute the lacIq promoter in front of YF1 and Fixk2.
 
In this part, we use the T7 promoter to substitute the lacIq promoter in front of YF1 and Fixk2.
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<p>
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<b><font size="3"> 0 hour:</font></b>
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<img src="https://2019.igem.org/wiki/images/thumb/c/c0/T--ShanghaiTech_China--characterization_25%28T7c_0h%29.png/447px-T--ShanghaiTech_China--characterization_25%28T7c_0h%29.png" alt="T7c_0h" width="395" height="635"></p>
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<p>
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<b><font size="3"> 6 hour:</font></b>
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<img src="https://2019.igem.org/wiki/images/thumb/f/f6/T--ShanghaiTech_China--characterization_26%28T7c_6h%29.png/800px-T--ShanghaiTech_China--characterization_26%28T7c_6h%29.png" alt="T7c_6h" width="790" height="635"></p>
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<p>
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<b><font size="3"> 12 hour:</font></b>
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<img src="https://2019.igem.org/wiki/images/thumb/2/23/T--ShanghaiTech_China--characterization_27%28T7c_12h%29.png/800px-T--ShanghaiTech_China--characterization_27%28T7c_12h%29.png" alt="T7c_12h" width="790" height="635"></p>
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<p>
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<b><font size="3"> 24 hour:</font></b>
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<img src="https://2019.igem.org/wiki/images/thumb/d/d9/T--ShanghaiTech_China--characterization_28%28T7c_24h%29.png/800px-T--ShanghaiTech_China--characterization_28%28T7c_24h%29.png" alt="T7c_24h" width="790" height="635"></p>
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<p>
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<b><font size="3"> 30 hour:</font></b>
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<img src="https://2019.igem.org/wiki/images/thumb/2/2b/T--ShanghaiTech_China--characterization_29%28T7c_30h%29.png/800px-T--ShanghaiTech_China--characterization_29%28T7c_30h%29.png" alt="T7c_30h" width="790" height="635"></p>
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<p>
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<b><font size="3"> 36 hour:</font></b>
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<img src="https://2019.igem.org/wiki/images/thumb/e/ed/T--ShanghaiTech_China--characterization_30%28T7c_36h%29.png/800px-T--ShanghaiTech_China--characterization_30%28T7c_36h%29.png" alt="T7c_36h" width="790" height="635"></p>
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<p>
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To see more changes on pDawn, go to the websites below.</p>
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<p>
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pDawn+mEGFP: BBa_K3278000</p>
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<p>
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pDawn with T7 promoter: BBa_3278001</p>
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<p>
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pDawn with mtac promoter: BBa_K3278002 (good)</p>
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<p>
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pdawn with tac promoter: BBa_K3278005</p>
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<p>
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pDawn with YF2: BBa_K3278006 (good)</p>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 02:53, 20 October 2019


pDawn with T7 promoter

pDawn is a blue light control system which can start the expression of target gene under the inducement of blue light. More information in BBa_K1075044 added by iGEM13_Bonn.

However, expression leakage and delay are the two main problems of pdawn system. Leakage refers to the expression of target protein without light. According to our measurement, samples strictly kept away from light still had significant expression of GFP after being cultured for one night. Therefore, we carefully studied the gene circuits of pdawn system [2] [3] and hypothesized that enhancing the strength of the promoter of YF1 may enhance the repression of protein expression in the dark, thus solving the leakage problem.

In this part, we use the T7 promoter to substitute the lacIq promoter in front of YF1 and Fixk2.

0 hour: <img src="447px-T--ShanghaiTech_China--characterization_25%28T7c_0h%29.png" alt="T7c_0h" width="395" height="635">

6 hour: <img src="800px-T--ShanghaiTech_China--characterization_26%28T7c_6h%29.png" alt="T7c_6h" width="790" height="635">

12 hour: <img src="800px-T--ShanghaiTech_China--characterization_27%28T7c_12h%29.png" alt="T7c_12h" width="790" height="635">

24 hour: <img src="800px-T--ShanghaiTech_China--characterization_28%28T7c_24h%29.png" alt="T7c_24h" width="790" height="635">

30 hour: <img src="800px-T--ShanghaiTech_China--characterization_29%28T7c_30h%29.png" alt="T7c_30h" width="790" height="635">

36 hour: <img src="800px-T--ShanghaiTech_China--characterization_30%28T7c_36h%29.png" alt="T7c_36h" width="790" height="635">


To see more changes on pDawn, go to the websites below.

pDawn+mEGFP: BBa_K3278000

pDawn with T7 promoter: BBa_3278001

pDawn with mtac promoter: BBa_K3278002 (good)

pdawn with tac promoter: BBa_K3278005

pDawn with YF2: BBa_K3278006 (good)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2231
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 109
    Illegal NgoMIV site found at 241
    Illegal NgoMIV site found at 335
    Illegal NgoMIV site found at 628
    Illegal NgoMIV site found at 1122
    Illegal NgoMIV site found at 1140
    Illegal NgoMIV site found at 1230
    Illegal AgeI site found at 460
    Illegal AgeI site found at 1588
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1683
    Illegal BsaI.rc site found at 571