Coding

Part:BBa_K933001:Design

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

leader sequence with threonine repeats. mCherry and GFP reporters


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 66
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 66
    Illegal NheI site found at 94
    Illegal NheI site found at 117
    Illegal NotI site found at 72
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 66
    Illegal BamHI site found at 197
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 66
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 66
    Illegal XbaI site found at 81
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was made by digesting out the orginal repeat sequence in BBa_K933002 at BamHI and HindIII and ligating in the threonine 6 repeat of ACT. The repeat sequence was made from annealed oligos.

Sequence data provided for: EcoRI, XbaI, J23100 Promoter, RBS34, leader sequence, BamHI, Threonine ACT repeat, HindIII

Construct.jpg

The part still has the Promoter, RBS34, leader sequence, repeat sequence (specifically Threonine ACT 6 repeats) mcherry, RBS30, and GFP. Built in sPB1C3 also.

Source

This piece was already partially assembled by a Ph.D student working in the Richard lab at Penn State named Mike Speer. Additional work was performed to perfect the construct with an alternate RBS from the registry. Repeat sequences were ordered as ssDNA oligos.

References