Intermediate

Part:BBa_K933002:Design

Designed by: Hannah Jepsen-Burger   Group: iGEM12_Penn_State   (2012-10-01)

short leader sequence with mCherry and GFP reporters


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 71
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 71
    Illegal NheI site found at 99
    Illegal NheI site found at 122
    Illegal NotI site found at 77
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 71
    Illegal BamHI site found at 202
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 71
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 71
    Illegal XbaI site found at 86
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

An alternative RBS site was used to reduce mCherry expression in this construct. There is an RBS30 site before the GFP so that the GFP will be expressed not depending on the mCherry expression. The repeat sequence that is inputted will cause the variation in RFP expression. The parts were constructed in the sPB1C3 vector as the backbone.


Only sequence provided was for: EcoR1, Xba1, J23100 Promoter, RBS34, leader sequence, BamHI, repeat sequence spot, HindIII

the rest of sequence is the mCherry and GFP

Source

This part was partially assembled by a Penn State Ph.D student in Dr. Thomas Richard's lab named Mike Speer. Completion of the codon optimization project was performed by iGEM members. An alternative RBS site was used to reduce mCherry expression.

References