Difference between revisions of "Part:BBa K1321328:Design"
Line 6: Line 6:
  
 
===Design Notes===
 
===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.
+
BBa_K1321302 was created by restricting BBa_K1033924 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.
  
  

Revision as of 21:48, 20 October 2014

J-sRNA-331Bb


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4541
    Illegal EcoRI site found at 1297
    Illegal SpeI site found at 1207
    Illegal PstI site found at 1685
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1297
    Illegal EcoRI site found at 4541
    Illegal SpeI site found at 1207
    Illegal PstI site found at 1685
    Illegal NotI site found at 1678
    Illegal NotI site found at 4547
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1297
    Illegal EcoRI site found at 4541
    Illegal XhoI site found at 1145
    Illegal XhoI site found at 1151
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4541
    Illegal EcoRI site found at 1297
    Illegal SpeI site found at 1207
    Illegal PstI site found at 1685
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4541
    Illegal EcoRI site found at 1297
    Illegal XbaI site found at 4556
    Illegal SpeI site found at 1207
    Illegal PstI site found at 1685
    Illegal NgoMIV site found at 2860
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1198
    Illegal BsaI.rc site found at 987


Design Notes

BBa_K1321302 was created by restricting BBa_K1033924 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