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Plasmid

Part:BBa_K1321201:Design

Designed by: Xenia Spencer-Milnes   Group: iGEM14_Imperial   (2014-10-09)


VHb in pSEVA331-Bb


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 3384
    Illegal suffix found in sequence at 513
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3384
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 514
    Illegal PstI site found at 528
    Illegal NotI site found at 521
    Illegal NotI site found at 3390
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3384
    Illegal BglII site found at 477
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3384
    Illegal suffix found in sequence at 514
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3384
    Illegal XbaI site found at 3399
    Illegal SpeI site found at 514
    Illegal PstI site found at 528
    Illegal NgoMIV site found at 1703
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

BBa_K12321201 was created by restricting BBa_K1321200 and BBa_K1321300 with PstI and XbaI, gel purifying the correct fragments, ligating using T4 DNA ligase, and transformed into chemically competent E.coli DH10B cells. Transformants were cultured on LB-chloramphenicol plates for 2 days at 37C inverted, and correct transformants were determnined via colony PCR, restriction digestion analysis and sequencing.

Source

BBa_K13421201 was created using BBa_K1321200 and BBa_K1321300.

References