Part:BBa_K5471012:Design
Cassette 1 (EF1a-mir195-eGFP-SV40pA)
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 460
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2009
Illegal PstI site found at 460
Illegal NotI site found at 2003 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 608
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 460
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 460
Illegal NgoMIV site found at 343
Illegal AgeI site found at 68 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1113
Design Notes
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). EF1a-mir195 infused promoter's design is described in detail in https://parts.igem.org/Part:BBa_K5471011 . Downstream of this composite promoter a kozak sequence and eGFP coding region were placed, for the production of a strong fluoerescent signal when the cassette is expressed. SV40pA signal was chosen for its reliability.
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_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_K5471009 (hsa-mir-195 genomic locus 150bp flank) https://parts.igem.org/Part:BBa_K5471011 (EF1a-mir195 infused promoter)