Difference between revisions of "Part:BBa K1321333:Design"
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
 
The AraC and pBAD elements were obtained by PCR, using the Biobricks BBa_K325108, BBa_K325218 and BBa_K325219 as DNA templates. The primers used during this procedure were as follows:
 
The AraC and pBAD elements were obtained by PCR, using the Biobricks BBa_K325108, BBa_K325218 and BBa_K325219 as DNA templates. The primers used during this procedure were as follows:
−
Forward: TACTAGTAGCGGCCGCTGCAG
+
<li>Forward: TACTAGTAGCGGCCGCTGCAG
−
Reverse: GCTAGCCCAAAAAAACGGGTATGGAG
+
<li>Reverse: GCTAGCCCAAAAAAACGGGTATGGAG
 
The resulting elements were DpnI-treated following a standard procedure and were submitted to self-ligation at room temperature for 1h+. The ligation mix was then transformed into chemically competent DH10B Escherichia coli using a standard chemical transformation protocol and plated out on Chloramphenicol-specific LB plates.
 
The resulting elements were DpnI-treated following a standard procedure and were submitted to self-ligation at room temperature for 1h+. The ligation mix was then transformed into chemically competent DH10B Escherichia coli using a standard chemical transformation protocol and plated out on Chloramphenicol-specific LB plates.
  

Revision as of 17:04, 21 October 2014

pSB-AraC-pBAD


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

The AraC and pBAD elements were obtained by PCR, using the Biobricks BBa_K325108, BBa_K325218 and BBa_K325219 as DNA templates. The primers used during this procedure were as follows:

  • Forward: TACTAGTAGCGGCCGCTGCAG
  • Reverse: GCTAGCCCAAAAAAACGGGTATGGAG The resulting elements were DpnI-treated following a standard procedure and were submitted to self-ligation at room temperature for 1h+. The ligation mix was then transformed into chemically competent DH10B Escherichia coli using a standard chemical transformation protocol and plated out on Chloramphenicol-specific LB plates.

    Source

    References

    https://parts.igem.org/Part:BBa_K325108 https://parts.igem.org/Part:BBa_K325218

    https://parts.igem.org/Part:BBa_K325219