Difference between revisions of "Part:BBa K3733005"
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===Usage and Biology===
 
===Usage and Biology===
 
<p>J23106 is a medium-strength constitutive promoter of the Anderson family. By editing J23106's -35 region by replacing nucleotides 3-5 from TAC to ATA, J23106a was successfully constructed (<b>Figure 1</b>)，which is a very weak promoter. </p>
 
<p>J23106 is a medium-strength constitutive promoter of the Anderson family. By editing J23106's -35 region by replacing nucleotides 3-5 from TAC to ATA, J23106a was successfully constructed (<b>Figure 1</b>)，which is a very weak promoter. </p>
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<p><b>Figure 1. </b> Modified methods of J23106a</p>
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<center><img src="https://static.igem.org/mediawiki/parts/9/99/T--HZAU-China--J23106a_1.png" style="width:793px;height:360px"></center>
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<center><b>Figure 1.</b>Modified methods of J23106a</center>
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===Functional Parameters===
 
===Functional Parameters===
 
<p>To characterize this part, J23106a and J23106 were cloned into pSC101 vector separately. We chose neGFP（https://parts.igem.org/Part:BBa_K3733012） as the reporter. Plasmids were transferred into <i>E.coli</i> DH5α. The E. coli strain was cultured overnight, and the relative intensity of J23106a was evaluated by measuring RFU/OD<sub>600</sub> under the control of two different promoters. The relative fluorescence unit (RFU) was determined by Synergy H1 microplate reader. </p>
 
<p>To characterize this part, J23106a and J23106 were cloned into pSC101 vector separately. We chose neGFP（https://parts.igem.org/Part:BBa_K3733012） as the reporter. Plasmids were transferred into <i>E.coli</i> DH5α. The E. coli strain was cultured overnight, and the relative intensity of J23106a was evaluated by measuring RFU/OD<sub>600</sub> under the control of two different promoters. The relative fluorescence unit (RFU) was determined by Synergy H1 microplate reader. </p>
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<p><b>Figure 2. </b> Comparison of relative strength between J23106 and J23106a.The microplate reader gain adopts 50</p>
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<center><img src="https://static.igem.org/mediawiki/parts/5/51/T--HZAU-China--J23106a_2.png" style="width:793px;height:360px"></center>
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<center><b>Figure 2.</b>Comparison of relative strength between J23106 and J23106a.The microplate reader gain adopts 50</center>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3733005 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3733005 SequenceAndFeatures</partinfo>

Revision as of 09:07, 15 October 2021


J23106a:Improved promoter of J23106

J23106a is a constitutive promoter obtained by mutating J23106.（https://parts.igem.org/Part:BBa_J23106） LThe strength of this promoter is much weaker than J23106.


Functional Parameters

To characterize this part, J23106a and J23106 were cloned into pSC101 vector separately. We chose neGFP（https://parts.igem.org/Part:BBa_K3733012） as the reporter. Plasmids were transferred into E.coli DH5α. The E. coli strain was cultured overnight, and the relative intensity of J23106a was evaluated by measuring RFU/OD600 under the control of two different promoters. The relative fluorescence unit (RFU) was determined by Synergy H1 microplate reader. 

无标题文档

Figure 2.Comparison of relative strength between J23106 and J23106a.The microplate reader gain adopts 50

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
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]