Difference between revisions of "Part:BBa K777000:Design"

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===Design Notes===
 
===Design Notes===
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Genomic sequence was amplified using the following primers.<br><br>
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* Genomic sequence was amplified using the following primers.
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Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
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** Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
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*** Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´
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Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´
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*** Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´
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<br>
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Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´
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* One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.  
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<br><br>
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** Primers QuikChange (QC): Primers were provided by SIGMA.
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<br><br>
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*** Fwd: TarQC for: 5´ ACTGATTGATTAcCTAGATTATGGCAATACTGGAG 3´
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One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR. <br><br>
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*** Rev: TarQC rev: 5´ TGCCATAATCTAGgTAATCAATCAGTTCAGTTAAC 3´<br>(the uncapitalized letter induces the mutation for removal of the XbaI site)
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Primers QuikChange: Primers were provided by SIGMA.
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−
 
+
−
Fwd: TarQC for: 5´ ACTGATTGATTACCTAGATTATGGCAATACTGGAG 3´
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−
 
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Rev: TarQC rev: 5´ TGCCATAATCTAGGTAATCAATCAGTTCAGTTAAC 3´
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===Source===
 
===Source===

Revision as of 13:15, 17 September 2012

Tar receptor


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1282
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 111


Design Notes

  • Genomic sequence was amplified using the following primers.
    • Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
      • Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´
      • Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´


  • One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.
    • Primers QuikChange (QC): Primers were provided by SIGMA.
      • Fwd: TarQC for: 5´ ACTGATTGATTAcCTAGATTATGGCAATACTGGAG 3´
      • Rev: TarQC rev: 5´ TGCCATAATCTAGgTAATCAATCAGTTCAGTTAAC 3´
        (the uncapitalized letter induces the mutation for removal of the XbaI site)

Source

The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).

References

  • Derr P., Boder E. and Goulian M. Changing the specificity of a bacterial chemoreceptor. J. Mol. Biol. 2006. 355:923–932.