Generator

Part:BBa_K103017:Design

Designed by: Michael Lower   Group: iGEM08_Warsaw   (2008-10-09)
Revision as of 15:06, 28 October 2008 by Smaegol (Talk | contribs) (→‎Design Notes)

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OmpA_linker_alpha_linker under Plac


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1321
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Preparation of BBa_K103016 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=5&arg0=10_October_2008&arg1=13_October_2008&arg2=14_October_2008&arg3=15_October_2008&arg4=16_October_2008&name=Preparation%20of%20OmpA_linker_alpha_linker%20under%20Plac%20(BBa_K103017) here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).

Alpha fragment was amplified from [http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha pACYC177+OmpA-omega-ΔA-alpha] using primers: 5' GG GAGCTCGCTACTTACTCTAGCTTCCCGG 3' and 5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC 3'.

Resulting product was used as a template for PCR with primers: 5' GG GAGCTCGCTACTTACTCTAGCTTCCCGG 3' and 5' ATACTAGTACCGGATCCAGAACCTCCTCCACCGCTACCTCCACCTCCAGAAC 3' (to obtain alpha_linker fragment), followed by digestion with SacI and BcuI.

Alpha_linker fragment was ligated into pSB2K3 vector containing lactose promoter and OmpA_linker (obtained by digest of pSB2K3 carrying BBa_K103018 with SacI and BcuI)

Source

References