Difference between revisions of "Part:BBa K660200:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K660200 short</partinfo>
 
<partinfo>BBa_K660200 short</partinfo>
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===Design Notes===
 
===Design Notes===
−
Illegal restriction sites - site directed mutagenesis
+
<h3>Illegal Restriction Sites</h3>
 +
 
 +
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/>
 +
 
 +
<u>1st PstI site:</u><br/>
 +
5': GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG<br/><br/>
 +
 
 +
&#160;&#160;&#160; Forward Primer: 5' CAG TTG CAG ACG GGT ATC TTG CAG TTT AAT TTC CGC<br/><br/>
 +
   
 +
&#160;&#160;&#160; Reverse Primer: 5' GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG<br/><br/>
 +
 
 +
MTP= 79.33  <br/><br/>
 +
 
 +
 
 +
<u>2nd PstI site:</u><br/>
 +
5': GCG ATT GAG CTG CAA AGC GGC AAT CCG C<br/><br/>
 +
 
 +
&#160;&#160;&#160; Forward Primer: 5' G CGG ATT GCC GCT TTG CAG CTC AAT CGC<br/><br/>
  
 +
&#160;&#160;&#160; Reverse Primer: 5' GCG ATT GAG CTG CAA AGC GGC AAT CCG C<br/><br/>
  
 +
MTP= 78.71<br/><br/>
  
 
===Source===
 
===Source===
  
−
Originally isolated from horse sweat.  
+
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. 
  
−
Received from researchers
 
  
 
===References===
 
===References===

Revision as of 13:05, 21 September 2011

Latherin (Surfactant Protein)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 200
    Illegal PstI site found at 431
    Illegal PstI site found at 569
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 200
    Illegal PstI site found at 431
    Illegal PstI site found at 569
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 514
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 200
    Illegal PstI site found at 431
    Illegal PstI site found at 569
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 200
    Illegal PstI site found at 431
    Illegal PstI site found at 569
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Illegal Restriction Sites

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.

1st PstI site:
5': GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG

    Forward Primer: 5' CAG TTG CAG ACG GGT ATC TTG CAG TTT AAT TTC CGC

    Reverse Primer: 5' GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG

MTP= 79.33


2nd PstI site:
5': GCG ATT GAG CTG CAA AGC GGC AAT CCG C

    Forward Primer: 5' G CGG ATT GCC GCT TTG CAG CTC AAT CGC

    Reverse Primer: 5' GCG ATT GAG CTG CAA AGC GGC AAT CCG C

MTP= 78.71

Source

Latherin was originally isolated from horse sweat. It was expressed in E.coli by researchers at University of Glasgow. We received it from them in a plasmid and amplified it up by PCR.


References