Generator

Part:BBa_K1598003:Design

Designed by: Marta Napiorkowska   Group: iGEM15_UCL   (2015-08-27)
Revision as of 21:05, 27 August 2015 by Martanap (Talk | contribs) (→‎Design Notes)

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RBS-ChAT-6xHis-Terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 682
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 303
    Illegal XhoI site found at 1770
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1095
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1765


Design Notes

- human CHAT gene (UniProt accession no: P28329) was codon-optimized for E.coli and provided with C-terminal 6xHis tag
- due to its length, the sequence was synthesized as two gBlock fragments with 30 bp overlap between each other and with 20 bp overlaps with respective PSB1C3 ends
- the construct was cloned into PSB1C3 using Gibson Assembly

Source

DNA synthesis

References