Difference between revisions of "Part:BBa K611025"
 
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<partinfo>BBa_K611025 short</partinfo>
 
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Parts K611021 through K611027 are variants of the wild type LacI repressible promoter, BBa_R0010. These parts were generated through three rounds of mutagenic PCR and each have around 1 to 7 mutations. Each of these variants differ in properties such as transcriptional strength and repressor binding affinity and have been well characterized. This can be very useful in tuning genetic circuits for specific outputs. These parts were characterized using K611013.
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Parts K611021 through K611027 are variants of the wild type LacI repressible promoter, BBa_R0010. These parts were generated through three rounds of mutagenic PCR and each have around 1 to 7 mutations. Each of these variants differ in properties such as transcriptional strength and repressor binding affinity and have been well characterized. This can be very useful in tuning genetic circuits for specific outputs. These parts were characterized using K611013. The data below was collected in strain BW22826 which is, among other things, ΔAraBAD and ΔLacI. For more detailed information on this strain, it can be found here: http://cgsc.biology.yale.edu/Strain.php?ID=92290
  
 
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<img src="https://static.igem.org/mediawiki/parts/f/fb/UCD_Plot_allmuts_crop.png" width="600">
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<img src="https://static.igem.org/mediawiki/parts/9/93/UCD_mut5_2D_crop.png" width="600">
 
<img src="https://static.igem.org/mediawiki/parts/e/ee/UCD_R10WT.png" width="450">
 
<img src="https://static.igem.org/mediawiki/parts/e/ee/UCD_R10WT.png" width="450">
 
<img src="https://static.igem.org/mediawiki/parts/5/53/UCD_R10mut5.png" width="450">
 
<img src="https://static.igem.org/mediawiki/parts/5/53/UCD_R10mut5.png" width="450">
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When induced with arabinose(as seen on the lower left axis), pBAD is derepressed allowing for transcription of the Lac repressor.  This is seen as a decrease in the reporter, GFP (on the left, vertical axis).  If IPTG is added(as seen on the right axis), it binds the Lac repressor which frees the promoter to transcribe.
  
 
https://static.igem.org/mediawiki/2011/0/0d/UCD_Partsreg_R10mut_seq.png
 
https://static.igem.org/mediawiki/2011/0/0d/UCD_Partsreg_R10mut_seq.png
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===Functional Parameters===
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==Functional Parameters: Austin_UTexas==
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<body>
 
<partinfo>BBa_K611025 parameters</partinfo>
 
<partinfo>BBa_K611025 parameters</partinfo>
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<h3><center>Burden Imposed by this Part:</center></h3>
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<center><img src = "https://static.igem.org/mediawiki/parts/f/fa/T--Austin_Utexas--no_burden_icon.png" style = "width:160px;height:120px"></center>
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<figcaption><center><b>Burden Value: 5.7 ± 6.1% </b></center></figcaption>
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<p> Burden is the percent reduction in the growth rate of <i>E. coli</i> cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the
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<a href="https://parts.igem.org/Part:BBa_K3174002">BBa_K3174002</a> - <a href="https://parts.igem.org/Part:BBa_K3174007">BBa_K3174007</a> pages for more information on the methods, an explanation of the sources of burden,  and other conclusions from a large-scale measurement project conducted by the <a href="http://2019.igem.org/Team:Austin_UTexas">2019 Austin_UTexas team</a>.</p>
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<p>This functional parameter was added by the <a href="https://2020.igem.org/Team:Austin_UTexas/Contribution">2020 Austin_UTexas team.</a></p>
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Latest revision as of 22:31, 3 September 2020

LacI Promoter Variant #5

Parts K611021 through K611027 are variants of the wild type LacI repressible promoter, BBa_R0010. These parts were generated through three rounds of mutagenic PCR and each have around 1 to 7 mutations. Each of these variants differ in properties such as transcriptional strength and repressor binding affinity and have been well characterized. This can be very useful in tuning genetic circuits for specific outputs. These parts were characterized using K611013. The data below was collected in strain BW22826 which is, among other things, ΔAraBAD and ΔLacI. For more detailed information on this strain, it can be found here: http://cgsc.biology.yale.edu/Strain.php?ID=92290


When induced with arabinose(as seen on the lower left axis), pBAD is derepressed allowing for transcription of the Lac repressor. This is seen as a decrease in the reporter, GFP (on the left, vertical axis). If IPTG is added(as seen on the right axis), it binds the Lac repressor which frees the promoter to transcribe.

UCD_Partsreg_R10mut_seq.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Functional Parameters: Austin_UTexas

BBa_K611025 parameters

Burden Imposed by this Part:

Burden Value: 5.7 ± 6.1%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods, an explanation of the sources of burden, and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.