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URL: http://www.addgene.org/11312
Proper Citation: RRID:Addgene_11312
Insert Name: 3 perfect binding sites for CXCR4 siRNA antisense
Bacterial Resistance: Ampicillin
Defining Citation: PMID:12600936
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:5256; Vector Backbone:pGL3-control; Vector Types:Luciferase; Bacterial Resistance:Ampicillin
Comments: The end of the ORF and the beginning of the 3' UTR is (with the TAA stop codon, followed by T, then the XbaI site TCTAGA): GATCGCCGTG TAA T TCTAGA GTCGGGGCG. We digested the plasmids with XbaI, and ligated in the following: ctaga ttccgagatatcggtaat gggcc. This produced a modified vector, still containing an XbaI site, but now also containing an ApaI site (GGGCCC), to allow for directional cloning of 3' UTR inserts. The final vector contains, starting with the stop codon TAA: TAA T Tctaga ttccgagatatcggtaat gggccC TAGA. All UTR inserts were then constructed in the following format: TCTAGACTCGAGCCGG[binding site]ATCGCGGGCCC. A single perfect binding site is AAGTTTTCACTCCAGCTAACACCGG. The CXCR4 siRNA used is GUUUUCACUCCAGCUAACACATTCAAAAGUGAGGUCGAUUGU.
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Source: Addgene
