Difference between revisions of "Part:BBa K2213007"
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This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible.
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This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible.<br>
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2213007 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K2213007 parameters</partinfo>
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Function:
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This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A medium strength Anderson promoter here, low strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213006), and a high strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213008).<br>
 
This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A medium strength Anderson promoter here, low strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213006), and a high strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213008).<br>
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The expression levels compared to low and high are as shown. As the medium strength promoter had similar expression levels to the high promoter, it was not expressed alongside any Eut subunits. This result was unexpected and should be taken into consideration when choosing either the medium or high promoter.<br>
 
The expression levels compared to low and high are as shown. As the medium strength promoter had similar expression levels to the high promoter, it was not expressed alongside any Eut subunits. This result was unexpected and should be taken into consideration when choosing either the medium or high promoter.<br>
 +
 +
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<!-- Add more about the biology of this part here
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===Usage and Biology===
 +
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2213007 SequenceAndFeatures</partinfo>
 +
 +
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K2213007 parameters</partinfo>
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<!-- -->

Revision as of 01:57, 1 November 2017


MediumPromoter_PduD(1-20)_mCherry

Manchesterigem17-med-tag1.png
Figure1. Circuit diagram of BBa_K2213007.

Improvements


This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible.

This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A medium strength Anderson promoter here, low strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213006), and a high strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213008).

The PduD tag was combined with the medium strength Anderson promoter (https://parts.igem.org/Part:BBa_J23108) and mCherry.

PromoterComparison800p.png

A gradient of fluorescence is evident when compared to low and high promoters. When expressed the distribution of the mCherry signal was homogeneous and fluorescence level was slightly less than the high promoter. .



TagExpression500p.jpg
The expression levels compared to low and high are as shown. As the medium strength promoter had similar expression levels to the high promoter, it was not expressed alongside any Eut subunits. This result was unexpected and should be taken into consideration when choosing either the medium or high promoter.


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]