Coding

Part:BBa_K660200:Design

Designed by: Glasgow Uni 2011   Group: iGEM11_Glasgow   (2011-09-16)
Revision as of 13:05, 21 September 2011 by HannahRalph (Talk | contribs)

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