Composite

Part:BBa_K3726073:Design

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-19)
Revision as of 14:04, 20 October 2021 by Sbarraga (Talk | contribs)

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L2_TOL_ABB


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1139
    Illegal EcoRI site found at 7738
    Illegal PstI site found at 798
    Illegal PstI site found at 3917
    Illegal PstI site found at 8640
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1139
    Illegal EcoRI site found at 7738
    Illegal NheI site found at 3646
    Illegal NheI site found at 7319
    Illegal NheI site found at 7342
    Illegal NheI site found at 8365
    Illegal PstI site found at 798
    Illegal PstI site found at 3917
    Illegal PstI site found at 8640
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1139
    Illegal EcoRI site found at 7738
    Illegal XhoI site found at 2860
    Illegal XhoI site found at 7774
    Illegal XhoI site found at 8011
    Illegal XhoI site found at 8793
    Illegal XhoI site found at 8913
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1139
    Illegal EcoRI site found at 7738
    Illegal PstI site found at 798
    Illegal PstI site found at 3917
    Illegal PstI site found at 8640
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1139
    Illegal EcoRI site found at 7738
    Illegal PstI site found at 798
    Illegal PstI site found at 3917
    Illegal PstI site found at 8640
    Illegal NgoMIV site found at 724
    Illegal NgoMIV site found at 5053
    Illegal NgoMIV site found at 6569
    Illegal AgeI site found at 3003
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 7498


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_K3726066” Lv1_ HspA-A, and “BBa_K3726071” Lv1_AcrbV2 - B are linked between themselves through 3'Con1 “BBa_K2560070” and 5'Con2 “BBa_K2560066” connectors. Likewise, “BBa_K3726071” Lv1_AcrbV2 - B and “BBa_K3726069” Lv1 SigB-B are linked between themselves through 3'Con2 “BBa_K2560071” and 5'Con3 “BBa_K2560067”connectors.

3'Con1 Is located in the downstream region of Lv1_ HspA-A and 5'Con2 is located in the upstream region of Lv1_AcrbV2 - B. Moreover, 3'Con2 is located in the downstream region of Lv1_AcrbV2 - B and 5'Con3 in the upstream region of Lv1 SigB-B.

Additionally, all this Lv2 composite part is flanked by two homology regions, included within the connector parts used for Lv.2 MoClo Assembly “BBa_K3726108” 5CON1(H)_NS3(mod)-up (PCC 11801) and “BBa_K3726109” 3CON5(H)_NS3(mod)-down (PCC 11801).

For last, regarding to CDS_lv0_SigB we have removed an internal BsmbI site in the base 862 in order to make this pat MoClo compatible with the marburg collection. However, the Sapl site should be removed in order to make this part RFC1000 compatible.


Source

This construct has been made by golden gate reaction.