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A dataset of ligand-dependent binding sites was generated using the SiteHopper create tool [13] where default parameters create a binding site patch within 4 Å of a specified bound ligand.
A genome-wide map of predicted STAT1 binding sites was generated as follows.
The plasmid pCAG-125b-6 pCAG-125b-6 pCAG-125b-6 copies of binding sites was generated using the isocaudamer of SpeI/ NheI.
In order to decrease endogenous miR-181a-5p miR-181a-5p miR-181a-5puct with four repeats containing miR-181 binding sites was generated.
To identify candidate TFs that can regulate multiple Hcrt-neuron specific genes, a map of possible TF binding sites was generated based on tHcrt-neuron specifictranscripts.
The plasmid containing site-directed mutations in putative p53 binding sites was generated by replacing the wild type Mlu I- Xho I fragment of pGL2-210 with a mutant fragment synthesized artificially pGL2-210BB, Table 1, Fig. 3).
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After formation of the dityrosine bond a complete set of well-formed zinc binding sites is generated.
All the designed mutant nucleotide sequences were re-analysed in Match 1.0 Public to check for loss of the specific transcription factor binding site and that no new binding sites were generated.
A binding p-value was then determined for each genomic position by Wilcoxon rank sum test and binding sites were generated from those more significant than specified thresholds with a maximum gap of 500 and minimum run of 350.
Two different types of sets of example binding sites were generated for each cutoff score from 2 to 7. They correspond to sequences drawn from a step-function distribution or a Boltzmann distribution, in each case using a maximum energy cutoff for selected sites.
Mutations in the miR-200c binding sites were generated by PCR directed mutagenesis.
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