Protein_Domain

Part:BBa_K103003:Design

Designed by: Michael Lower   Group: iGEM08_Warsaw   (2008-09-14)
Revision as of 21:33, 26 October 2008 by Smaegol (Talk | contribs) (→‎Design Notes)

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 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.

Resulting fragment - B Domain of Staphylococcal protein A was cloned into into pACYC177 vector to generate pACYC177+OmpA-omega-ΔA-alpha vector, used by our team in 'hunter and prey' tests.

BBa K103003 part was prepared by PCR on pACYC177+OmpA-omega-ΔA-alpha vector using AL_BXNE (5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC 3') and APSacSpe (5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC 3') primers, followed by digestion with EcoRI and BcuI

Source

References