Difference between revisions of "Part:BBa K143006:Design"

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<partinfo>BBa_K143006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K143006 SequenceAndFeatures</partinfo>
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===Design Notes===
 
===Design Notes===
  
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The EpsE integration sequences were designed from the EpsE (aka YveO) gene sequence<cite>#1</cite> and identification of the sequence directly upstream of the gene on the chromosome (found using NCBI's sequence viewer). The upstream and EpsE gene sequence was analysed for restriction sites and primers (with biobrick prefix and suffix sequences) for two approximately equally sized integration sequences were desgined. The integration sequences were then produced by PCR cloning with Pfu DNA polymerase
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The EpsE integration sequences were based upon the EpsE (aka YveO) gene sequence<cite>#1</cite> and the sequence of the upstream region on the chromosome (found using NCBI's sequence viewer). The upstream and EpsE gene sequence was analysed for restriction sites and primers (with biobrick prefix and suffix sequences) for two approximately equally sized integration sequences were desgined. The integration sequences were then produced by PCR cloning with Pfu DNA polymerase
  
 
===Source===
 
===Source===

Revision as of 13:28, 18 September 2008

3' Integration sequence for the EpsE locus of B. subtilis



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 211
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The EpsE integration sequences were based upon the EpsE (aka YveO) gene sequence#1 and the sequence of the upstream region on the chromosome (found using NCBI's sequence viewer). The upstream and EpsE gene sequence was analysed for restriction sites and primers (with biobrick prefix and suffix sequences) for two approximately equally sized integration sequences were desgined. The integration sequences were then produced by PCR cloning with Pfu DNA polymerase

Source

The 3’ integration sequence was taken from the B.subtilis chromosome and is homologous to the middle section of the EpsE gene. It was produced by PCR cloning with Pfu DNA polymerase

References

<biblio>

  1. 1 pmid=9384377

</biblio>