Plasmid

Part:BBa_K4686061

Designed by: Maia Weatherly   Group: iGEM23_Princeton   (2023-10-10)


pPB_Gal4UAS_tBFP_PGK_mCitrine

A piggybac vector that constitutively expresses mCitrine for selection after transfection. It also has a UAS site that drives expression of tBFP when Gal4 is present. It works alongside part BBa_K4686062, which is a receptor that is able to release Gal4 upon binding. This plasmid is intended to serve as a reporter for that receptor.

Uses primers BBa_K4686015, BBa_K4686016, BBa_K4686017, BBa_K4686018, BBa_K4686019, and BBa_K4686020 to amplify pHR_Gal4UAS_tBFP_PGK_mCitrine and pBR70. pHR_Gal4UAS_tBFP_PGK_mCitrine is from Kole Roybal, and pBR70 was given to us by a mentor.

This plasmid confers Carbenicillin resistance to the bacteria for selection while cloning. It was assembled via Gibson Assembly.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 175
    Illegal EcoRI site found at 1658
    Illegal XbaI site found at 312
    Illegal XbaI site found at 1146
    Illegal XbaI site found at 5881
    Illegal SpeI site found at 307
    Illegal SpeI site found at 336
    Illegal SpeI site found at 1404
    Illegal SpeI site found at 3110
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 175
    Illegal EcoRI site found at 1658
    Illegal NheI site found at 360
    Illegal NheI site found at 6209
    Illegal SpeI site found at 307
    Illegal SpeI site found at 336
    Illegal SpeI site found at 1404
    Illegal SpeI site found at 3110
    Illegal NotI site found at 1097
    Illegal NotI site found at 3374
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 175
    Illegal EcoRI site found at 1658
    Illegal BglII site found at 6223
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 175
    Illegal EcoRI site found at 1658
    Illegal XbaI site found at 312
    Illegal XbaI site found at 1146
    Illegal XbaI site found at 5881
    Illegal SpeI site found at 307
    Illegal SpeI site found at 336
    Illegal SpeI site found at 1404
    Illegal SpeI site found at 3110
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 175
    Illegal EcoRI site found at 1658
    Illegal XbaI site found at 312
    Illegal XbaI site found at 1146
    Illegal XbaI site found at 5881
    Illegal SpeI site found at 307
    Illegal SpeI site found at 336
    Illegal SpeI site found at 1404
    Illegal SpeI site found at 3110
    Illegal NgoMIV site found at 2972
    Illegal AgeI site found at 1269
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4550
    Illegal BsaI.rc site found at 1004
    Illegal SapI.rc site found at 5632


[edit]
Categories
Parameters
None