Difference between revisions of "Part:BBa K1045013:Design"
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===Design Notes===
 
===Design Notes===
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For cloning of [[Part:BBa_K1045013|BBa_K1045013]], first, oligos for [[Part:BBa_K1045012|BBa_K1045012]] were hybridized. The hybridization oligos were designed such that their double-stranded hybridization product would correspond to the sequence of [[Part:BBa_K1045012|BBa_K1045012]] cut with ''EcoRI'' and ''SpeI'' at the prefix and suffix sites. Second, this hybridization product was ligated in front of the GFP generator [[Part:BBa_E0240|BBa_E0240]] previously cut with ''EcoRI'' and ''XbaI''.
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For cloning of '''BBa_K1045013''', first, oligos for [[Part:BBa_K1045012|BBa_K1045012]] were hybridized. The hybridization oligos were designed such that their double-stranded hybridization product would correspond to the sequence of [[Part:BBa_K1045012|BBa_K1045012]] cut with ''EcoRI'' and ''SpeI'' at the prefix and suffix sites. Second, this hybridization product was ligated in front of the GFP generator [[Part:BBa_E0240|BBa_E0240]] previously cut with ''EcoRI'' and ''XbaI''.
  
 
===Source===
 
===Source===
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The hybridization oligos mentioned under "Design Notes" were purchased from Sigma-Aldrich. The sequence originated from the Parts Registry page of [[Part:BBa_J23110|BBa_J23110]] and the sequence of the DarR operator found upstream of ''darR'' (Zhang ''et al.'', 2013). The plasmid of part [[Part:BBa_E0240|BBa_E0240]] derived from the distribution kit 2013.
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To be continued
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===References===
 
===References===
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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

Latest revision as of 16:49, 26 October 2013

Promoter - DarR operator - GFP generator BBa_E0240


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 728


Design Notes

For cloning of BBa_K1045013, first, oligos for BBa_K1045012 were hybridized. The hybridization oligos were designed such that their double-stranded hybridization product would correspond to the sequence of BBa_K1045012 cut with EcoRI and SpeI at the prefix and suffix sites. Second, this hybridization product was ligated in front of the GFP generator BBa_E0240 previously cut with EcoRI and XbaI.

Source

The hybridization oligos mentioned under "Design Notes" were purchased from Sigma-Aldrich. The sequence originated from the Parts Registry page of BBa_J23110 and the sequence of the DarR operator found upstream of darR (Zhang et al., 2013). The plasmid of part BBa_E0240 derived from the distribution kit 2013.

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