Difference between revisions of "Part:BBa K137060"

 
 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K137060 short</partinfo>
 
<partinfo>BBa_K137060 short</partinfo>
Line 5: Line 4:
 
A promoter facing upstream is flanked by two FimE binding sites. GFP is downstream to this fimE switch. Active FimE will flip the promoter such that GFP is expresssed.
 
A promoter facing upstream is flanked by two FimE binding sites. GFP is downstream to this fimE switch. Active FimE will flip the promoter such that GFP is expresssed.
  
−
<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
 +
The series K137057-K137062 were placed in pSB4K5 and cotransformed with pFIP<sup>1</sup>. Using the Caltech iGEM [http://2008.igem.org/Team:Caltech/Protocols/FimE FimE assay], the series of parts produced the following fluorescence values:
 +
 +
[[Image:FimE.gif]]
 +
 +
Error bars show the standard deviation of experiments performed in triplicate.
 +
 +
Maximum fluorescence was observed to occur at about a separation length of 250 bp. This distance may be a function of entropy and enthalpy. The chances that two DNA segments on the same strand come together statistically decrease as the two segments are separated by intervening sequences. In addition, two sites very close together are unlikely to come into contact with each other, as it is energetically unfavorable to bend DNA to such a great degree. These two opposing forces might explain the presence of an optimal separation length.
  
−
<!-- -->
+
# Ham TS, Lee SK, Keasling JD, and Arkin AP. A tightly regulated inducible expression system utilizing the fim inversion recombination switch. Biotechnol Bioeng 2006 May 5; 94(1) 1-4.
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K137060 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K137060 SequenceAndFeatures</partinfo>

Latest revision as of 01:12, 22 October 2008

GFP fimE switch with 450 bp tetR promoter

A promoter facing upstream is flanked by two FimE binding sites. GFP is downstream to this fimE switch. Active FimE will flip the promoter such that GFP is expresssed.

Usage and Biology

The series K137057-K137062 were placed in pSB4K5 and cotransformed with pFIP1. Using the Caltech iGEM [http://2008.igem.org/Team:Caltech/Protocols/FimE FimE assay], the series of parts produced the following fluorescence values:

FimE.gif

Error bars show the standard deviation of experiments performed in triplicate.

Maximum fluorescence was observed to occur at about a separation length of 250 bp. This distance may be a function of entropy and enthalpy. The chances that two DNA segments on the same strand come together statistically decrease as the two segments are separated by intervening sequences. In addition, two sites very close together are unlikely to come into contact with each other, as it is energetically unfavorable to bend DNA to such a great degree. These two opposing forces might explain the presence of an optimal separation length.

  1. Ham TS, Lee SK, Keasling JD, and Arkin AP. A tightly regulated inducible expression system utilizing the fim inversion recombination switch. Biotechnol Bioeng 2006 May 5; 94(1) 1-4.

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 1206