Part:BBa_K5471013:Design
Cassette 2 (ITR-EF1a-mir195-eGFP-SV40pA-ITR)
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 587
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2136
Illegal PstI site found at 587
Illegal NotI site found at 2130 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 735
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 587
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 587
Illegal NgoMIV site found at 470
Illegal AgeI site found at 195 - 1000INCOMPATIBLE 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)