Coding
LigA

Part:BBa_K525740:Design

Designed by: Anna Drong   Group: iGEM11_Bielefeld-Germany   (2011-09-12)
Revision as of 21:58, 21 September 2011 by Adrong (Talk | contribs) (→‎Design Notes)

DNA ligase from Escherichia coli (LigA) with His-tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1422
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1636


Design Notes

  • This is the complete sequence of NAD+ -dependent DNA ligase.
  • It is designed in RFC 25
    • NgoMIV restriction site downstream the start codon to easily fuse an N-terminal domain to the coding sequence

Source

  • PCR on the Escherichia coli TOP 10 DNA cloned into BioBrick Vectors
  • Primers with BioBrick Freiburg prefix in the fwd primer. Suffix and His-tag in the rev primer.
    • fwd: 5'-acgtgaattcgcggccgcttctagatggccggcgaatcaatcgaacaacaactgac-3'
    • rev: 5'-acgtctgcagcggccgctactagtaatgatgatgatgatgatggctacccagcaaacgc-3'

References