Plasmid

Part:BBa_K4686069:Design

Designed by: Janae Gordon   Group: iGEM23_Princeton   (2023-10-12)


pMCherry+AmylaseDegrader_Link_EGF_mEGFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 2456
    Illegal XbaI site found at 1382
    Illegal PstI site found at 7959
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 2456
    Illegal PstI site found at 7959
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 2456
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 2456
    Illegal XbaI site found at 1382
    Illegal PstI site found at 7959
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 2456
    Illegal XbaI site found at 1382
    Illegal PstI site found at 7959
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1637
    Illegal BsaI.rc site found at 7785
    Illegal SapI site found at 6702


Design Notes

We designed this plasmid so that the Amylase Degrader signal peptide would facilitate the secretion of the EGF ligand so that we could test for secretion efficiency. We also had to consider overhangs for the signal peptide insert into the backbone so that they could be assembled with Gibson Assembly.



Source

pMLB_mCherry plasmid sourced fromPiet de Boer T7_PB_SFFV_CD4_EGFf_mEGFP_TEVs_CD28_ plasmid sourced from Beatrice Ramm Amylase Degrader sequence sourced from UniProt


References