Difference between revisions of "Part:BBa K1045000:Design"

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We noticed that, when cloning the DarR operator via hybridization oligos, it was difficult to obtain transformants indicating that potential secondary structures that might be formed by the palindromic sequences of these hybridization oligos interfere with the transformation. The palindromic sequences could result from the sequence of the DarR operator itself.
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We noticed that, when cloning the DarR operator via hybridization oligos, it was difficult to obtain transformants. The DarR operator harbors a palindromic sequence which could form secondary structures. Such secondary structures might account for the inefficient transformations.
  
 
===Source===
 
===Source===

Latest revision as of 07:31, 27 October 2013

DarR operator


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]


Design Notes

Vector.jpg

We noticed that, when cloning the DarR operator via hybridization oligos, it was difficult to obtain transformants. The DarR operator harbors a palindromic sequence which could form secondary structures. Such secondary structures might account for the inefficient transformations.

Source

The sequence of this part corresponds to the sequence of the DarR operator upstream of darR, which was published by Zhang et al. in 2013. The part was generated using hybridization oligos purchased from Sigma-Aldrich.

[http://www.google.de/imgres?um=1&hl=de&biw=1433&bih=691&tbm=isch&tbnid=-_tTmC4jAFeHFM:&imgrefurl=http://2007.igem.org/wiki/index.php/Paris/transclusion_example&docid=4kRb6plD_PfdGM&imgurl=https://static.igem.org/mediawiki/2007/d/d2/Pbluescript_SK-.jpg&w=316&h=241&ei=x8ZCUsOgPMrg4QSExIGoCQ&zoom=1&iact=hc&vpx=427&vpy=229&dur=1608&hovh=192&hovw=252&tx=52&ty=213&page=1&tbnh=161&tbnw=199&start=0&ndsp=34&ved=1t:429,r:3,s:0,i:89 Vector iGEM 2007 Wiki]

References

Lei Zhang, Weihui Li, and Zheng-Guo He (2013) “DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in Mycobacterium smegmatis”, THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 288, NO. 5, pp. 3085–3096