Generator

Part:BBa_J36849:Design

Designed by: Perry Tsai   Group: iGEM06_Harvard   (2006-10-29)
Revision as of 21:11, 29 October 2006 by Perrytsai (Talk | contribs) (→‎Design Notes)


Lac-inducible generator of Lpp-OmpA(46-66)-Streptavidin single-chain dimeric + His6 tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1179
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1188
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 609
    Illegal AgeI site found at 1143
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

There was originally a XbaI site TCTAGA at nucleotides 118-123 of the part, which was mutated to AGCCGC. It is missing its start codon at the beginning, as the intention was to fuse this with surface display domains upstream.

There are no spacer nucleotides between the part and the XbaI restriction site, or between the part and the SpeI restriction site, so that the mixed site would be 6bp long, and reading frame would be maintained between protein domains.

Source

PCR off a plasmid obtained from Alice Ting's lab at MIT.

References