Composite

Part:BBa_K1321329:Design

Designed by: Michael Florea   Group: iGEM14_Imperial   (2014-10-08)

GFP-LVA under TetR control in pSEVA331-Bb


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4748
    Illegal suffix found in sequence at 1877
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4748
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 1878
    Illegal PstI site found at 1892
    Illegal NotI site found at 1885
    Illegal NotI site found at 4754
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4748
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4748
    Illegal suffix found in sequence at 1878
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4748
    Illegal XbaI site found at 4763
    Illegal SpeI site found at 1878
    Illegal PstI site found at 1892
    Illegal NgoMIV site found at 3067
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1616


Design Notes

BBa_K1321302 was created by restricting part BBa_K145280 and BBa_K1321300 (pSEVA331-BB backbone) with XbaI and PstI, gel purifying the resulting fragments and ligating with T4 ligase. Ligated DNA was then transformed into chemically competent cells, screened via colony PCR and culture PCR, and confirmed by sequencing.


Source

References