Composite

Part:BBa_K3726058:Design

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-20)
Revision as of 13:27, 20 October 2021 by Sbarraga (Talk | contribs)

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Lv2_BOH1_ABC


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1850
    Illegal PstI site found at 18
    Illegal PstI site found at 1755
    Illegal PstI site found at 2003
    Illegal PstI site found at 3988
    Illegal PstI site found at 4237
    Illegal PstI site found at 7832
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1850
    Illegal NheI site found at 6540
    Illegal NheI site found at 6563
    Illegal NheI site found at 6838
    Illegal NheI site found at 7066
    Illegal PstI site found at 18
    Illegal PstI site found at 1755
    Illegal PstI site found at 2003
    Illegal PstI site found at 3988
    Illegal PstI site found at 4237
    Illegal PstI site found at 7832
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1850
    Illegal BglII site found at 8166
    Illegal BglII site found at 8283
    Illegal BglII site found at 10702
    Illegal BamHI site found at 4320
    Illegal BamHI site found at 4659
    Illegal XhoI site found at 5568
    Illegal XhoI site found at 9386
    Illegal XhoI site found at 10229
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1850
    Illegal PstI site found at 18
    Illegal PstI site found at 1755
    Illegal PstI site found at 2003
    Illegal PstI site found at 3988
    Illegal PstI site found at 4237
    Illegal PstI site found at 7832
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1230
    Illegal EcoRI site found at 1850
    Illegal PstI site found at 18
    Illegal PstI site found at 1755
    Illegal PstI site found at 2003
    Illegal PstI site found at 3988
    Illegal PstI site found at 4237
    Illegal PstI site found at 7832
    Illegal NgoMIV site found at 1494
    Illegal NgoMIV site found at 1594
    Illegal NgoMIV site found at 1900
    Illegal NgoMIV site found at 4915
    Illegal AgeI site found at 594
    Illegal AgeI site found at 2272
    Illegal AgeI site found at 3443
    Illegal AgeI site found at 4443
  • 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_K3726044” BOH1_A_LV1, and “BBa_K3726045” BOH1_B_LV1 are linked between themselves through 3'Con1 K2560070 and 5'Con2 BBa_K2560066 connectors. Likewise, “BBa_K3726045” BOH1_B_LV1 and “BBa_K3726046” BOH1_C_LV1 are linked between themselves through 3'Con2 “BBa_K2560071” and 5'Con3 “BBa_K2560067”connectors.

3'Con1 Is located in the downstream region of BOH1_A_LV1 and 5'Con2 is located in the upstream region of BOH1_B_LV1. Moreover, 3'Con2 is located in the downstream region of BOH1_B_LV1 and 5'Con3 in the upstream region of BOH1_C_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 ”

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.