Difference between revisions of "Part:BBa K731040:Design"

 
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<partinfo>BBa_K731040 short</partinfo>
 
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===Design Notes===
 
===Design Notes===
Primers for Gibson's Assembly were designed to add a linker before sfGFP and remove the stop codons from CysE M256I.
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This part was created by Gibson assembly starting from K731030. sfGFP was amplified by PCR from K515005, removing the RBS. CysE and sfGFP are separated by a short nucleotide linker that encodes for the amino acidic sequence GSGSGS.
 
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===Source===
 
===Source===
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===References===
 
===References===
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Jones-Mortimer, M.C.(1968). The control of sulphate reduction in Escherichia coli by O-acetyl-l-serine. Biochem. J. 106, 33. <br>
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Kredich & Tomkins, 1966. The enzymic synthesis of L-cysteine in Escherichia coli and Salmonella typhimurium. Biochem. J. 241, 4955-4965.  <br>
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Denk, D., and A. Bock. 1987. L-cysteine biosynthesis in Escherichia coli: nucleotide sequence and expression of the serine acetyltransferase (cysE) gene from the wild-type and a cysteine-excreting mutant. J. Gen. Microbiol. 133:515–525.  <br>

Latest revision as of 21:29, 26 October 2012

Inducible araC-pBAD promoter regulating a sfGFP-tagged M256I CysE


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2045
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1909
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI.rc site found at 2075


Design Notes

This part was created by Gibson assembly starting from K731030. sfGFP was amplified by PCR from K515005, removing the RBS. CysE and sfGFP are separated by a short nucleotide linker that encodes for the amino acidic sequence GSGSGS.

Source

K731030

References

Jones-Mortimer, M.C.(1968). The control of sulphate reduction in Escherichia coli by O-acetyl-l-serine. Biochem. J. 106, 33.
Kredich & Tomkins, 1966. The enzymic synthesis of L-cysteine in Escherichia coli and Salmonella typhimurium. Biochem. J. 241, 4955-4965.
Denk, D., and A. Bock. 1987. L-cysteine biosynthesis in Escherichia coli: nucleotide sequence and expression of the serine acetyltransferase (cysE) gene from the wild-type and a cysteine-excreting mutant. J. Gen. Microbiol. 133:515–525.