Coding
LigA

Part:BBa_K525730:Design

Designed by: Anna Drong   Group: iGEM11_Bielefeld-Germany   (2011-09-12)
Revision as of 21:57, 21 September 2011 by Adrong (Talk | contribs) (→‎Design Notes)

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DNA ligase from Escherichia coli (LigA) with PT7 and RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1449
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2047
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1663


Design Notes

  • This is the complete sequence of NAD+ -dependent DNA ligase with T7-promoter and RBS.
  • It is designed in RFC 25 assembly standard
    • AgeI restriction site upstream of stop codon to easily fuse a C-terminal domain to the coding sequence

Source

  • PCR on the Escherichia coli TOP 10 DNA cloned into BioBrick Vectors
  • Primers with BioBrick prefix, T7-promoter and RBS in the fwd primer. Freiburg suffix in the rev primer.
    • fwd: 5'-acgtgaattcgcggccgcttctagagtaatacgactcactatagggaaagaggagaaaaatggaatcaatcgaacaac-3'
    • rev: 5'-acgtctgcagcggccgctactagtattaaccggtgctacccagcaaacgc-3'

References