Device

Part:BBa_K1172908:Design

Designed by: Tore Bleckwehl   Group: iGEM13_Bielefeld-Germany   (2013-09-19)

Part 1 of the Biosafety-System Lac of Growth (pRha lacI alr)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1623
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1547
    Illegal BamHI site found at 2249
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1665
    Illegal AgeI site found at 1965
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1422


Design Notes

  • Amplification of the rhamnose promoter PRha (BBa_K194004) with its pSB1C3 backbone using specific Gibson-Primer:
pRhA_rev (41 bp)

5'-TTGTTTCTCCTCTTTCTAGTATTCATTACGACCAGTCTAAA-3'

pSB1C3_rha_suf (42 bp)

5'-GTGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAG-3'
  • Amplification of the repressor lacI with Gibson-Primer containing the RBS (BBa_B0034):
lacI_fwd (45 bp)

5'-TACTAGAAAGAGGAGAAACAAATGGTGAATGTGAAACCAGTAACG-3'

lacI_rev (22 bp)

5'-TTATTAAGCTACTAAAGCGTAG-3'


alr_lacI (67 bp)

5'‑CTACGCTTTAGTAGCTTAATAATACTAGAAAGAGGAGAAATACTAGATGCAAGCGGCAACTGTTGTG-3'

alr_lacI_2 (42 bp)

5'-CTGCAGCGGCCGCTACTAGTA-3'


  • Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].



Source


References