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URL: http://www.addgene.org/20793
Proper Citation: RRID:Addgene_20793
Insert Name: mir-206-1/mir-133b
Organism: Danio rerio
Bacterial Resistance: Ampicillin
Defining Citation: PMID:19240133
Vector Backbone Description: Backbone Marker:Bartel Lab; Backbone Size:0; Vector Backbone:pBSII SK+ modified to introduce GFP and more cloning sites; Vector Types:zebrafish expression; Bacterial Resistance:Ampicillin
Comments: For polycistronic mir-206-1/mir-133b, we were not able to identify the transcriptional start precisely by 5′-RACE due to the repetitive sequence in this region. Analyses of EST clones (BM259633, CF999541, CF348604, DR729161) mapped the approximate start of the mir-206-1/mir-133b primary transcript to 1.4 kb from the mir-206 hairpin. A 4.5-kb upstream fragment that includes ∼0.9 kb of the primary transcript was amplified from genomic DNA using primers 3 and 4 (5'-CTGCCCCATTTAGACTCTTTCTTGACG-3' and 5'-CCAGAAGAACCCAGACGAAACCCAC-3'). The promoter fragment was fused to a GFP reporter gene. The miRNA promoter fusion was flanked by I-SceI meganuclease recognition sites to increase the efficiency of transgenensis. Please note that Addgene's sequencing results show some differences from the published sequence. The depositing lab is aware of the differences.
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Source: Addgene
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