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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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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Preparation of BBa_K103003 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=18_September_2008&arg1=19_September_2008&arg2=22_September_2008&arg3=23_September_2008&arg4=24_September_2008&arg5=25_September_2008&arg6=26_September_2008&name=Preparation%20of%20%26Delta%3BA%20%28BBa_K103003%29 here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).
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B-domain of Staphylococcal protein A was amplified from [http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha pACYC177+OmpA-omega-&Delta;A-alpha] vector using primers: 5' TAGAATTCGCGGCCGCTTCTAGA GGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3' and 5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3'. Resulting fragment was then digested with EcoRI and BcuI and cloned into standard BioBrick vector [https://parts.igem.org/wiki/index.php?title=Part:pSB1A3 pSB1A3]
  
 
===Source===
 
===Source===

Latest revision as of 13:15, 28 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

Preparation of BBa_K103003 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=18_September_2008&arg1=19_September_2008&arg2=22_September_2008&arg3=23_September_2008&arg4=24_September_2008&arg5=25_September_2008&arg6=26_September_2008&name=Preparation%20of%20%26Delta%3BA%20%28BBa_K103003%29 here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).

B-domain of Staphylococcal protein A was amplified from [http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha pACYC177+OmpA-omega-ΔA-alpha] vector using primers: 5' TAGAATTCGCGGCCGCTTCTAGA GGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC 3' and 5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC 3'. Resulting fragment was then digested with EcoRI and BcuI and cloned into standard BioBrick vector pSB1A3

Source

References