Plasmid

Part:BBa_K1321302:Design

Designed by: Michael Florea   Group: iGEM14_Imperial   (2014-10-08)
Revision as of 11:24, 10 October 2014 by Mf1512 (Talk | contribs) (→‎Design Notes)


pSEVA331-BB with J23100-RFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 3776
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3776
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
    Illegal NotI site found at 913
    Illegal NotI site found at 3782
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3776
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3776
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3776
    Illegal XbaI site found at 3791
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
    Illegal NgoMIV site found at 2095
    Illegal AgeI site found at 616
    Illegal AgeI site found at 728
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

BBa_K1321302 was created by restricting BBa_J23100 (Anderson promoter+RFP insert) 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

y

References