Difference between revisions of "Part:BBa K103003:Design"

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===Design Notes===
 
===Design Notes===
 
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B Domain of Staphylococcal protein A was incidentally amplified from vector carrying full protein A sequnce using primers AP+NotI (5' AA GCGGCCGC C GGTTGACTTCCCCGCGGAATTC 3') and AL+link10+homo2 (5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC 3'). This primers should be able to generate both: full-length protein A-coding sequence and only B-domain coding sequence.<br><br>
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B Domain of Staphylococcal protein A was amplified from vector carrying TapTag sequence (pDRIVE+TapTag) using AL_BXNE (5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3') and APSacSpe (5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3') primers, followed by digestion with EcoRI and BcuI
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Resulting fragment - B Domain of Staphylococcal protein A was cloned into into pACYC177 vector to generate <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA-omega-&Delta;A-alpha</a> vector, used by our team in 'hunter and prey' tests. <br><br>
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BBa K103003 part was prepared by PCR on <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA-omega-&Delta;A-alpha</a> vector using AL_BXNE (5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3') and APSacSpe (5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3') primers, followed by digestion with EcoRI and BcuI
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Revision as of 22:15, 26 October 2008

B domain of Staphylococcal protein A


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 91
    Illegal BamHI site found at 9
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

B Domain of Staphylococcal protein A was amplified from vector carrying TapTag sequence (pDRIVE+TapTag) using AL_BXNE (5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC 3') and APSacSpe (5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC 3') primers, followed by digestion with EcoRI and BcuI

Source

References