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

Revision as of 12:32, 19 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

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. When we used hybridization oligos for BBa_K1045012 (a promoter sequence followed by the DarR operator) we did not observe these problems. Consequently, it might be recommendable to clone the DarR operator as a part of bigger sequences.

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

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 [[File:vector.jpg|200px]]
-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. When we used hybridization oligos for [Part:BBa_K1045012|BBa_K1045012] (a promoter sequence followed by the DarR operator) we did not observe these problems. Consequently, it might be recommendable to clone the DarR operator as a part of bigger sequences.
+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. When we used hybridization oligos for [[Part:BBa_K1045012|BBa_K1045012]] (a promoter sequence followed by the DarR operator) we did not observe these problems. Consequently, it might be recommendable to clone the DarR operator as a part of bigger sequences.
 ===Source===