Difference between revisions of "Part:BBa K581010"
 
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Regulatory proteins for receiving signaling of CAI-1.
 
Regulatory proteins for receiving signaling of CAI-1.
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<partinfo>BBa_K581010 parameters</partinfo>
 
<partinfo>BBa_K581010 parameters</partinfo>
 
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===Design Notes===
 
===Design Notes===
  
 
▪ A silent mutation (A->G) at base 1086 was made to remove PstI site in the region of cqsS gene
 
▪ A silent mutation (A->G) at base 1086 was made to remove PstI site in the region of cqsS gene
 +
 
▪ A silent mutation (A->G) at base 1650 was made to remove EcoRI site in the region of cqsS gene
 
▪ A silent mutation (A->G) at base 1650 was made to remove EcoRI site in the region of cqsS gene
 +
 
▪ A silent mutation (G->A) at base 3234 was made to remove PstI site in the region of luxU gene
 
▪ A silent mutation (G->A) at base 3234 was made to remove PstI site in the region of luxU gene
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▪ cqsS and luxO were amplified by PCR
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▪ cqsS and luxO were amplified by PCR from the genome DNA of ''Vibrio cholera''
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▪ promoter Bba_J23106 + luxU was amplified by DNAWorks
 
▪ promoter Bba_J23106 + luxU was amplified by DNAWorks
  
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===Source===
 
===Source===
  
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▪ We received the genome DNA of Vibrio cholera  from Chinese Center for Disease Control and Prevention
+
▪ We received the genome DNA of ''Vibrio cholera'' from Chinese Center for Disease Control and Prevention
  
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▪ Although our system includes several genes from Vibrio cholera, these genes are not directly involved with virulence or antibiotic resistance in any case.
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▪ Although our system includes several genes from ''Vibrio cholera'', these genes are not directly involved with virulence or antibiotic resistance in any case.
  
  
 
===References===
 
===References===
  
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* [http://nar.oxfordjournals.org/content/30/10/e43.full.pdf+html].
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* [http://nar.oxfordjournals.org/content/30/10/e43.full.pdf+html] David M. Hoover, Jacek Lubkowski. DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis. ''Nucl. Acids Res'' 30 (10): e4 (2002).

Latest revision as of 16:11, 5 October 2011

promoter(BBa_J23106)-RBS-luxU-RBS-cqsS-RBS-luxO

Regulatory proteins for receiving signaling of CAI-1.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 4021
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 4021
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3096
    Illegal BamHI site found at 404
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 4021
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 4021
    Illegal AgeI site found at 2374
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 964
    Illegal SapI site found at 3284


Design Notes

▪ A silent mutation (A->G) at base 1086 was made to remove PstI site in the region of cqsS gene

▪ A silent mutation (A->G) at base 1650 was made to remove EcoRI site in the region of cqsS gene

▪ A silent mutation (G->A) at base 3234 was made to remove PstI site in the region of luxU gene

▪ cqsS and luxO were amplified by PCR from the genome DNA of Vibrio cholera

▪ promoter Bba_J23106 + luxU was amplified by DNAWorks


Source

▪ We received the genome DNA of Vibrio cholera from Chinese Center for Disease Control and Prevention

▪ Although our system includes several genes from Vibrio cholera, these genes are not directly involved with virulence or antibiotic resistance in any case.


References

  • [http://nar.oxfordjournals.org/content/30/10/e43.full.pdf+html] David M. Hoover, Jacek Lubkowski. DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis. Nucl. Acids Res 30 (10): e4 (2002).