Project

Part:BBa_K3972004:Design

Designed by: Ingrid Kolen, Daniek Hoorn, Werner Doensen   Group: iGEM21_TU-Eindhoven   (2021-09-20)


Tetrathionate sensor with transferred TtrR and pTtrB185-269 activated ARG1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1658
    Illegal EcoRI site found at 4811
    Illegal XbaI site found at 1409
    Illegal PstI site found at 404
    Illegal PstI site found at 4668
    Illegal PstI site found at 4773
    Illegal PstI site found at 5151
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1658
    Illegal EcoRI site found at 4811
    Illegal NheI site found at 1543
    Illegal PstI site found at 404
    Illegal PstI site found at 4668
    Illegal PstI site found at 4773
    Illegal PstI site found at 5151
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1658
    Illegal EcoRI site found at 4811
    Illegal BglII site found at 2162
    Illegal BamHI site found at 5567
    Illegal XhoI site found at 5255
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1658
    Illegal EcoRI site found at 4811
    Illegal XbaI site found at 1409
    Illegal PstI site found at 404
    Illegal PstI site found at 4668
    Illegal PstI site found at 4773
    Illegal PstI site found at 5151
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1658
    Illegal EcoRI site found at 4811
    Illegal XbaI site found at 1409
    Illegal PstI site found at 404
    Illegal PstI site found at 4668
    Illegal PstI site found at 4773
    Illegal PstI site found at 5151
    Illegal NgoMIV site found at 2613
    Illegal NgoMIV site found at 3015
    Illegal AgeI site found at 334
    Illegal AgeI site found at 5534
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Insert designed and ordered from IDT. To make this part restriction and ligation reactions were performed, using the insert from IDT and the plasmid pET28a-T7-ARG1 ordered from Addgene. The ordered plasmid was not BioBrick compatible. Therefore, first mutate the sequence to make it TypeIIS BioBrick compatible. Use the following mutations: GvpC codon 25, GvpG codon 78, GvpL codon 241. Earlier mutation attempts failed.

Source

Insert ordered from IDT and pET28a-T7-ARG1 ordered from Addgene.

References