Part:BBa_K5471020:Design
pAAV-EF1a-mir195-eGFP-SV40pA
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1079
Illegal BamHI site found at 1040
Illegal XhoI site found at 3453 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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