Sharp Lab
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We started with two luciferase vectors, pGL3-control and pRL-TK, purchased from Promega.  The end of the ORF and the beginning of the 3' UTR is shown below, with the TAA stop codon in red, and an underlined XbaI site:

pGL3:
GATCGCCGTGTAATTCTAGAGTCGGGGCG pRL-TK:
AAATGAACAATAATTCTAGAGCGGCCGCT
  We digested the plasmids with XbaI, and ligated in the following: ctagattccgagatatcggtaatgggcc
  This produced a modified vector, still containing an XbaI site, but now also containing an ApaI site, to allow for directional cloning of 3' UTR inserts. Final vectors
TAATTctagattccgagatatcggtaatgggccCTAGA
 
 

All UTR inserts were then constructed in the following format:
TCTAGACTCGAGCCGG[binding site]ATCGCGGGCCC
 
 

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