Composite

Part:BBa_K5471013:Design

Designed by: Michail Diamantakis   Group: iGEM24_Patras-Med   (2024-09-30)


Cassette 2 (ITR-EF1a-mir195-eGFP-SV40pA-ITR)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 587
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2136
    Illegal PstI site found at 587
    Illegal NotI site found at 2130
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 735
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 587
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 587
    Illegal NgoMIV site found at 470
    Illegal AgeI site found at 195
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1240


Design Notes

The sequence of this cassette is derived from the direct cloning of Cassette 1 https://parts.igem.org/Part:BBa_K5471012 in between the pair of ITR sequences.


Source

This composite part was designed by our team for the purposes of our experiments. This part was designed by us, taking inspiration from "sc AAV miR26a eGFP" https://www.addgene.org/21894/ sc AAV miR26a eGFP was a gift from Joshua Mendell (Addgene plasmid # 21894 ; http://n2t.net/addgene:21894 ; RRID:Addgene_21894)

The individual basic parts used to build the cassette are as follows; https://parts.igem.org/Part:BBa_K5471021 (AAV2 ITR 5' end) https://parts.igem.org/Part:BBa_K5471005 (EF1a promoter and intron) https://parts.igem.org/Part:BBa_K5471006 (Kozak sequence) https://parts.igem.org/Part:BBa_K5471007 (eGFP coding region) https://parts.igem.org/Part:BBa_K5471008 (SV40pA signal) https://parts.igem.org/Part:BBa_K5471022 (AAV2 ITR 3'end) https://parts.igem.org/Part:BBa_K5471009 (hsa-mir-195 genomic locus 150bp flank) https://parts.igem.org/Part:BBa_K5471011 (EF1a-mir195 infused promoter) https://parts.igem.org/Part:BBa_K5471012 (Cassette 1 EF1a-mir195-eGFP-SV40pA) https://parts.igem.org/Part:BBa_K5471004 (AAV2 ITRs left and right)


References