From 2007.igem.org

Lab schedule

• mouse insulin gene cloning (delay to 8/7)
  • primer design for [http://www.informatics.jax.org/searches/accession_report.cgi?id=MGI%3A96573 insulin II of Mus musculus], [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?cmd=Retrieve&db=Nucleotide&list_uids=142350788&dopt=GenBank&WebEnv=04iJq7mV5tF1N_LSoJWAy8uc-0i_vXvfkGc3KMxcxnrIPG9K-LDVOuYkZWjzh6BpujGcDGNGQk0k9I%40D4770B146B72B990_0041SID&WebEnvRq=1 MN_008387.3 transcript]
    • sequence for standard assembly (marked in green) and PCR reaction (marked in red)
    • primer for A chain
      • translation (90-110): GIVDQCCTSICSLYQLENYCN
      • transcript (341-403): GGCATTGTAGATCAGTGCTGCACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAAC, 63 bp
      • left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATGATTGTAGATCAGTGCTGCAC
      • right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGTTGCAGTAGTTCTCCAGCT
    • primer for B chain
      • translation (25-54): FVKQHLCGSHLVEALYLVCGERGFFYTPMS
      • transcript (146-235):TTTGTCAAGCAGCACCTTTGTGGTTCCCACCTGGTGGAGGCTCTCTACCTGGTGTGTGGGGAGCGTGGCTTCTTCTACACACCCATGTCC, 90 bp
      • left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTTTGTCAAGCAGCACCTTTG
      • right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGGACATGGGTGTGTAGAAGA
• purchase of BCRC human insulin clone (ordered not paid)
• cutting sites of iGEM 2007 parts
  • ompR binding ompC promoter
    • ompR binding site (C1, C2, C3) extraction
    • assembly to insulin
  • [http://partsregistry.org/Part:BBa_C0082 tar-EnvZ fusion construct]: cut EnvZ part for our fusion construct
• antibiotics LB plate preparation
  • ampicillin
  • kanamycin