Difference between revisions of "Part:BBa K3971024"
 
(2 intermediate revisions by one other user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K3971024 short</partinfo>
 
<partinfo>BBa_K3971024 short</partinfo>
  
−
This composite part has been designed to characterize the activity of the native E.coli csc operon bidirectional promoter.
+
This composite part has been designed to characterize the activity of the native <i>E.coli</i> csc operon bidirectional promoter from the strain EC3132 [1]. In order to allow for modularity in the promoter region of the cassette, two restriction sites Nde1 and BamH1 flank it, so that the promoter can be easily excised out and replaced with another and the cassette can be used to characterize that promoter.
  
−
[[File:T--IISER-Pune-India--bidirectionalcassette.png|thumb|700x900px|centre|Figure of the composite construct for characterizing cscB transport efficiencies under different promoters. NS1a and NS1b are the two homology arms to Neutral Site 1.]]
+
A YFP and a BFP gene flank the promoter on both sides on opposite coding strands, and the relative fluorescence intensity can be used as a proxy for the strength of the promoter in both directions. Both fluorescence promoters have a double terminator (BBa_B0015).
 +
 +
 
 +
[[File:T--IISER-Pune-India--bidirectionalcassette.png|thumb|700x900px|centre|Figure of the composite construct for characterizing the activity of the native <i>E.coli</i> csc operon bidirectional promoter.]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 13: Line 16:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3971024 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3971024 SequenceAndFeatures</partinfo>
 +
 +
===References:===
 +
 +
[1] Jahreis, K., Bentler, L., Bockmann, J., Hans, S., Meyer, A., Siepelmeyer, J., & Lengeler, J. W. (2002). Adaptation of sucrose metabolism in the Escherichia coli wild-type strain EC3132. Journal of bacteriology, 184(19), 5307–5316. https://doi.org/10.1128/JB.184.19.5307-5316.2002
  
  

Latest revision as of 00:17, 22 October 2021


Promoter characterization cassette for native E.coli csc operon bidirectional promoter.

This composite part has been designed to characterize the activity of the native E.coli csc operon bidirectional promoter from the strain EC3132 [1]. In order to allow for modularity in the promoter region of the cassette, two restriction sites Nde1 and BamH1 flank it, so that the promoter can be easily excised out and replaced with another and the cassette can be used to characterize that promoter.

A YFP and a BFP gene flank the promoter on both sides on opposite coding strands, and the relative fluorescence intensity can be used as a proxy for the strength of the promoter in both directions. Both fluorescence promoters have a double terminator (BBa_B0015).


Figure of the composite construct for characterizing the activity of the native E.coli csc operon bidirectional promoter.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1699

References:

[1] Jahreis, K., Bentler, L., Bockmann, J., Hans, S., Meyer, A., Siepelmeyer, J., & Lengeler, J. W. (2002). Adaptation of sucrose metabolism in the Escherichia coli wild-type strain EC3132. Journal of bacteriology, 184(19), 5307–5316. https://doi.org/10.1128/JB.184.19.5307-5316.2002