BBa_K410000 1 BBa_K410000 Periplasmic Heat Generation 2010-10-26T11:00:00Z 2015-05-08T01:12:25Z The part consists of three sub-parts with varying origins. The cold-shock promoter, HybB, was amplified using PCR from the biobrick BBa_J43001 in the Spring 2010 Distribution. OmpA was amplified from a commercial vector pFLAGS-CTS (Sigma), a plasmid that contains the 81 bp signal peptide. Aox1a, the alternative oxidase from Sacred Lotus, was synthesized commercially by Mr. Gene, Co. The sequence was taken from Genbank (GenBank: AB491175.1) and is from the Sacred Lotus (Nelumbo Nucifera). What it is: This part consists of hybB-ompA-AOX1A. HybB is a cold shock promoter. OmpA is a periplasmic targeting sequence, and AOX1A is a gene responsible for thermogenesis in the sacred lotus, a plant. What it does: Induces AOX1A expression in response to a cold shock. Here, we describe a "cold shock" as incubating E. coli between 20-30C for 10-12 hours. The part should produce a thermogenic response raising the temperature of the E. coli's immediate environment (plate or liquid cultures) above ambient temperature. false false _522_ 0 7842 9 It's complicated true Design considerations revolved primarily around targeting the AOX to the periplasm in order to localize AOX near its proper substrates. false Christina Graves, Scott Holmes annotation2111604 1 AoxA (Alternative Oxidase) range2111604 1 483 1517 annotation2111710 1 NotI Site range2111710 1 394 401 annotation2111736 1 XmaI Site range2111736 1 477 482 annotation2111504 1 HybB Cold Shock Promoter range2111504 1 1 393 annotation2111563 1 OmpA range2111563 1 414 476 annotation2111580 1 Ribosome Binding Site range2111580 1 402 407 BBa_K410000_sequence 1 cgccgctatggactggataaagagatggtaatggatttctttcgtgagaataattcctgttctacgttgcgcttttttatggccggttatcgcctcgaaaattgatcaaacatacgtattatcttgctttaattaattacactaatgcttcttcccttcgttttagcgccccgccgcagtatcatgatatcgataaccataataaatgtgtggtaaatggcgcatcgatcgcattattgattttgcgattgaggcaaaatatatgccaggtcttcgcaacggaataactataaatgactggagataacaccctcatccattctcacggcattaaccgtcgtgatttcatgaagctttgtgcagcattagccgccaccatggggttaagtagcagcggccgcaggagatatcatatgaaaaagacagctatcgcgattgcagtggcactggctggtttcgctaccgttgcgcaagctcccgggatgatgaacagcaaactggcagctctgctgctgaaacaactgggttctgctaccgttcgtaccgtgacaatgggtcctctgaatgggattaccactgagtcctcgtgctttctgcatgcatctggtccggtcgttccaggtcgccgtacttggattcgcttttcttgtctgggagtccgtaatggttctaccagtgccctgaataacaaagagaaagaggagaaaggcgtccgtacaagttctactgttggtggcgccaatcgccctgaggataaaatgatcgtgtcctattggggtatgcctccggcaaatctgactaaaaaagacggctccgagtggaaatggaactcattccgtccatgggaaacctataaagccgatctgtcgatcgatctgaaaaaacaccatagcccggtgacgtttatggataaactggcctattggaccgttaaagccctgcgctatccgactgatattctgttccaaaaccgttatggctgtcgtgccatgatgctggagactgtggcggcagtaccagggatggttggggggatgctgctgcacctgaaatctctgcgtcgcttcgaacattctggtgggtggatcaaaacactgctggaagaagccgaaaacgagcgtatgcacctgatgaccttcatggaagtgtctcagccaaaatggtatgaacgtgccctggttgtagcagttcagggtgtgttcttcaacacgtattttctgggttatctgatcagccctcgcttcgctcatcgtgtggttggatatctggaggaggaagcaattcactcctataccgagtttctgaaagaactggacaaaggcaacatccagaacgtcccagcaccagcaattgccgtggattattggcaactgccaccggattctacgctgcgtgatgtggtgatggtcgtccgtgctgatgaagcacatcatcgtgacgtgaaccactttgccagcgacattcacgaccaaggctatgagctgaaagagtctcctgctcctctgggatatcattaa igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z