Generator

Part:BBa_K103020:Design

Designed by: Michael Lower   Group: iGEM08_Warsaw   (2008-10-09)
Revision as of 16:03, 28 October 2008 by Smaegol (Talk | contribs) (→‎Design Notes)

omega_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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 356


Design Notes

Preparation of BBa_K103019 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=25_September_2008&arg1=29_September_2008&arg2=30_September_2008&arg3=9_October_2008&arg4=10_October_2008&arg5=13_October_2008&arg6=14_October_2008&name=Preparation%20of%20omega_linker%20under%20PT7%20%28BBa_K103020%29 here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).

Omega_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' CAGCCATATGCTACTTACTCTAGCTTCCCGG 3' and 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 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).

Omega_linker under control of lactose promoter was amplified using primers: 5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAA TTAATACG 3' and 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 3' . PCR product were digested with EcoRI and BcuI and ligated into pSB2K3

Source

References