Plasmid

Part:BBa_K5471020:Design

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


pAAV-EF1a-mir195-eGFP-SV40pA


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1079
    Illegal XbaI site found at 1050
    Illegal PstI site found at 1022
    Illegal PstI site found at 3305
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1079
    Illegal NheI site found at 4854
    Illegal PstI site found at 1022
    Illegal PstI site found at 3305
    Illegal NotI site found at 4848
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1079
    Illegal BamHI site found at 1040
    Illegal XhoI site found at 3453
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1079
    Illegal XbaI site found at 1050
    Illegal PstI site found at 1022
    Illegal PstI site found at 3305
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1079
    Illegal XbaI site found at 1050
    Illegal PstI site found at 1022
    Illegal PstI site found at 3305
    Illegal NgoMIV site found at 3188
    Illegal AgeI site found at 2913
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2395
    Illegal BsaI site found at 3958
    Illegal SapI.rc site found at 767


Design Notes

This plasmid is comprised of the original pUC57 vector provided by Genscript, and the Cassette 2 https://parts.igem.org/Part:BBa_K5471013 cloned into position 2710. The production of the whole plasmid pAAV-EF1a-mir195-eGFP-SV40pA was made possible by Genscript

Source

This plasmid 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 production of the whole plasmid pAAV-EF1a-mir195-eGFP-SV40pA was made possible by Genscript


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), https://parts.igem.org/Part:BBa_K5471013 (Cassette 2 ITR-EF1a-mir195-eGFP-SV40pA-ITR) https://parts.igem.org/Part:BBa_K5471003 (pUC57 vector)

References

The production of the whole plasmid pAAV-EF1a-mir195-eGFP-SV40pA was made possible by Genscript