BBa_K808027 1 BBa_K808027 EstA : inactive fusion protein for bacterial surface expression 2012-09-01T11:00:00Z 2015-05-08T01:13:26Z Our parts derives from an established inactive EstA mutant on pEST100 vector, which was used in the following paper: Becker, S., Theile, S., Heppeler, N., Michalczyk, A., Wentzel, A., Wilhelm, S., Jaeger, K.-E., et al. (2005). A generic system for the Escherichia coli cell-surface display of lipolytic enzymes. FEBS letters, 579(5), 1177???82. doi:10.1016/j.febslet.2004.12.087 This Esterase A is a inactivated membrane bound esterase from Pseudomonas putida. Its active site is mutated so it shows no more hydrolytic activity. It can be used for cell surface display like its shown in the following paper: Taek Ho Yang, Jae Gu Pan, Yeon Soo Seo, Joon Shick Rhee Use of Pseudomonas putida EstA as an Anchoring Motif for Display of a Periplasmic Enzyme on the Surface of Escherichia coli Appl Environ Microbiol. 2004 December; 70(12): 6968???6976. doi: 10.1128/AEM.70.12.6968-6976.2004 false false _1065_ 0 11792 9 In stock false This certain variant of EstA should be used for cell surface expression and displaying of almost every kind of enzyme. There is no ATG at its 5' because its an C-terminal fusion protein. If you want to use you need to design it in the following way: N - Peri plasmatic signal sequence - Protein of interest - EstA - C We did not use any kind of BioBrick Assembly due to the generation of stop codons. If you would like to fuse EstA to any kind of protein, please use SOE PCR or an alternative kind of assembly strategy. false Marie Burghard, Henrik Cordes, Jascha Diemer, Adrian Eilingsfeld, Sven Jager, Philipp Rottmann, Rene Sahm, Arne Wehling BBa_K808027_sequence 1 gctccttcgccctattcgacgctggtcgtgttcggcgacgtccaggcccaacaagccgccggtcgcctggtggatagcgtgcaggccctgcaccaggccggcgcgcgctacatcgtggtctggctgttgcccgacctgggcctgaccccggctaccttcggtggtcccttgcagcctttcgccagccaactcagcggcgcgttcaacgccgagccgaccgcccagttgagccaggccggcgccaacgtcattccgttgaacatcccgctgctgctcaaggaaggcatggccaacccggcttccttcggcctggccgccgaccagaacctgatcggcacctgtttcagcggcaacggctgcaccatgaacccgacctacgggatcaacggcagcacgcccgacccgagcaaattgctgttcaacgacagcgtgcacccgaccatcaccggccagcgcctgatcgccgactacacctattcgctgctgtcggcgccctgggagctgaccctgctgccggaaatggcccacggcaccctgcgtgcctaccaggacgaactgcgcagccagtggcaggcggactgggagaactggcagaacgtcggccagtggcgcggcttcgtcggcggcggtggccagcgcctggacttcgactcccaggacagcgccgccagcggcgacggcaacggctacaacctgacccttggtggcagctaccgcatcgacgaggcctggcgcgccggggtcgccgccggtttctaccggcagaagctggaagccggcgccaaggattccgactaccggatgaacagctacatggccagcgccttcgtgcagcaccaggaaaaccgctggtgggccgacgcggcgttgaccggcggctacctcgactacgacgacctgaagcgcaagttcgccctgggcggcggcgagcgcagcgagaaaggcgacaccaacggccacctgtgggcgttcagcgcgcgcctgggctacgacatcgcccagcaggccgacagtccctggcacctgtcgccgttcgtcagcgccgactatgcacgggtcgaggtcgacggctattccgagaagggcgccagcgccaccgcgctcgactacgacgaccagaagcgcagctcgaagcgcctgggcgccggcctgcaaggcaagtacgcgttcggcagcgatacccagctgttcgccgagtacgcccacgaacgtgagtacgaggacgacacccaggacctgaccatgtccctcaacagcctgccgggcaatcgcttcaccctcgaaggctacaccccgcaggaccatctcaaccgcgtctcactcggcttcagccagaagctggcaccggagctgtcgctgcgcggcggctacaactggcgcaagggcgaggacgatacccagcagagcgtcagcctggcgctgagcctggacttctaataa 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