Generator

Part:BBa_K103019:Design

Designed by: Michael Lower   Group: iGEM08_Warsaw   (2008-10-09)
Revision as of 15:49, 28 October 2008 by Smaegol (Talk | contribs) (→‎Design Notes)

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alpha_linker under PT7


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Preparation of BBa_K103019 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=9&arg0=25_September_2008&arg1=9_October_2008&arg2=10_October_2008&arg3=13_October_2008&arg4=14_October_2008&arg5=16_October_2008&arg6=17_October_2008&arg7=20_October_2008&arg8=21_October_2008&name=Preparation%20of%20alpha_linker%20under%20PT7%20%28BBa_K103019%29 here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).

Alpha_linker 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' CGGG CATATG CACCCAGAAACGCTGGTGAAAG 3' and 5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC 3'. Product was digested with NdeI and SacI and ligated into pET15b vector ([http://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega pET15b-OmpA-omega] with previously removed XbaI site, cutted with NdeI and SacI).

Alpha_linker under control of lactose promoter was amplified using primers: 5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG 3' and 5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC 3'. PCR product were digested with EcoRI and SacI and ligated into pSB1A3 (obtained by digest of standard pSB1A3 carrying BBa_K103003 part.

Source

References