Composite

Part:BBa_K3726060:Design

Designed by: Santiago Barrag�n Ariza   Group: iGEM21_MADRID_UCM   (2021-10-20)


L2_BOH2A-GSG


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1873
    Illegal PstI site found at 18
    Illegal PstI site found at 1778
    Illegal PstI site found at 2026
    Illegal PstI site found at 3327
    Illegal PstI site found at 6974
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1873
    Illegal NheI site found at 5654
    Illegal NheI site found at 5677
    Illegal NheI site found at 5980
    Illegal NheI site found at 6208
    Illegal PstI site found at 18
    Illegal PstI site found at 1778
    Illegal PstI site found at 2026
    Illegal PstI site found at 3327
    Illegal PstI site found at 6974
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1873
    Illegal BglII site found at 7303
    Illegal BglII site found at 7420
    Illegal BglII site found at 9839
    Illegal BamHI site found at 3410
    Illegal BamHI site found at 3749
    Illegal XhoI site found at 4682
    Illegal XhoI site found at 8523
    Illegal XhoI site found at 9366
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1873
    Illegal PstI site found at 18
    Illegal PstI site found at 1778
    Illegal PstI site found at 2026
    Illegal PstI site found at 3327
    Illegal PstI site found at 6974
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1873
    Illegal PstI site found at 18
    Illegal PstI site found at 1778
    Illegal PstI site found at 2026
    Illegal PstI site found at 3327
    Illegal PstI site found at 6974
    Illegal NgoMIV site found at 1517
    Illegal NgoMIV site found at 1617
    Illegal NgoMIV site found at 1923
    Illegal NgoMIV site found at 4005
    Illegal AgeI site found at 594
    Illegal AgeI site found at 2295
    Illegal AgeI site found at 3533
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The part is a MoClo Lv.2 part, assembled in the acceptor Lv2 entry vector whose ori is “BBa_K2560036”, and it's kanamycin resistance cassette is “BBa_K2560133” . In accordance with the Marburg Collection MoClo design, the parts “BBa_K3726047” BOH2_A_LV1 and “BBa_K3726052” BOH1_C_GSG_LV1 are linked between themselves through 3'Con2 “BBa_K2560071” and 5'Con3 “BBa_K2560067”connectors. 3'Con2 is located in the downstream region of BOH2_A_LV1 and 5'Con3 in the upstream region of BOH1_C_GSG_LV1. Additionally, all this Lv2 composite part is flanked by two homology regions, included within the connector parts used for Lv.2 MoClo Assembly “BBa_K3726104” 5CON1(H)_NS1(mod)-up (PCC 11801) and “BBa_K3726105 ” 3CON5(H)_NS1(mod)-down (PCC 11801) that allows the double homologous recombination within the genome of PCC 11801.

Source

This construct has been made through golden gate reaction.

References

X. Liu, R. Miao, P. Lindberg y P. Lindblad, “Modular engineering for efficient photosynthetic biosynthesis of 1-butanol from CO2 in cyanobacteria", 2021.