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| | <partinfo>BBa_K660201 short</partinfo> | | <partinfo>BBa_K660201 short</partinfo> |
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| − | <partinfo>BBa_K660201 SequenceAndFeatures</partinfo>
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| − | ===Design Notes===
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| − | We are aware of two illegal restriction sites that exist in our latherin biobrick. Due to time constraints, we were unable to complete site directed mutagenesis. The primers we designed for this are below, for any future team.<br/><br/>
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| − | <u>1st PstI site:</u><br/>
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| − | 5': GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG<br/><br/>
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| − |     Forward Primer: 5' CAG TTG CAG ACG GGT ATC TTG CAG TTT AAT TTC CGC<br/><br/>
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| − |     Reverse Primer: 5' GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG<br/><br/>
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| − | MTP= 79.33 <br/><br/>
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| − | <u>2nd PstI site:</u><br/>
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| − | 5': GCG ATT GAG CTG CAA AGC GGC AAT CCG C<br/><br/>
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| − |     Forward Primer: 5' G CGG ATT GCC GCT TTG CAG CTC AAT CGC<br/><br/>
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| − |     Reverse Primer: 5' GCG ATT GAG CTG CAA AGC GGC AAT CCG C<br/><br/>
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| − | MTP= 78.71<br/><br/>
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| − | ===Source===
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| − | Latherin was originally isolated from horse sweat. It was expressed in <i>E.coli</i> by researchers at University of Glasgow. We received it from them in a plasmid and amplified it up by PCR to add a biobrick prefix and suffix and a 6xHIS tag.
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| − | ===References===
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| − | Beegley J, et al., 1986. Isolation and characterization of latherin, a surface-active protein from horse sweat<br/>http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=search&term=3753435>.
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| − | <br/><br/>
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| − | Kennedy M., 2011., Latherin and other biocompatible surfactant proteins<br/>
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| − | http://www.biochemsoctrans.org/bst/039/bst0391017.htm<br/><br/>
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| − | Information on "Affinity Purification" <br/>
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| − | http://www.brunschwig-ch.com/pdf/downloads/ABT_RapidRun_Procedure.pdf?PHPSESSID=cb79f5862da62a654bb1a6aefe87c62b
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