BBa_K1991007 1 BBa_K1991007 Pcons-RBS-LO-BamHI 2016-10-11T11:00:00Z 2016-10-12T03:28:23Z E. coli (Lpp-OmpA) A study showed that Lpp-OmpA (LO) hybrid can direct the heterologous protein GFP to the external surface of E. coli (Enzyme Microb Technol. 2001). Bacillus lipase (J Microbiol. 2014) and Fungi xylanase (Curr Microbiol. 2015) were demonstrated to be displayed on the cell surface of E. coli and maintained the functional enzyme activities. We???d like to display the AOX enzyme on bacterial surface by fusing with Lpp-OmpA and apply to the electrochemical analyzer by depositing on the test strip. false false _2458_ 31417 31417 9 false No false Chen, Pei-En component2493570 1 BBa_K1991004 component2493567 1 BBa_J23101 component2493569 1 BBa_B0034 annotation2493570 1 BBa_K1991004 range2493570 1 62 502 annotation2493567 1 BBa_J23101 range2493567 1 1 35 annotation2493569 1 BBa_B0034 range2493569 1 44 55 BBa_K1991004 1 BBa_K1991004 Lpp-OmpA-BamHI 2016-10-09T11:00:00Z 2016-10-12T03:03:40Z Modified based on BBa_K1694002 (Lpp-OmpA-N) Lpp and OmpA are outer membrane proteins of E. coli. Lpp-OmpA (LO) hybrid can direct heterologous proteins to bacterial cell surface. In 2015, NCTU-Formosa used it to display scFv (single chain fragment variable) antibodies on the surface of E. coli. They found that a fusion protein cannot be possible to be created under the standard BioBrick assembly rule, that is EcoRI(E)-XbaI(X)-GENE-SpeI(S)-PstI(P). The A part of EX-LO-SP and the B part of EX-scFV-SP, for example, are connected by cutting and ligation of SpeI plus PstI for the A part and XbaI plus PstI for the B part. The SCAR generated by XbaI/SpeI (ACTAGA) will form a stop codon just in front of the ATG start codon of the scFV protein of the B part. This situation has been officially mentioned by the BioBrick standard assembly. Therefore, in 2015, NCTU-Formosa created a novel part (BBa_K1694002) putting NcoI site between the LO part and SpeI site. However, when considering cloning, we found that an extra NcoI site is present on Cm resistance gene making it difficult be a vector for gene cloning. In 2016, Mingdao improved the part by replacing NcoI site with BamHI site (BBa_K1991004). Also, we???ve confirmed and prove the function of LO directing a fusion protein to the cell surface with enzyme activity in our project. false false _2458_ 31417 31417 9 false BBa_K1991004: Lpp-OmpA-BamHI/pSB1C3 The DNA fragment of Lpp-OmpA was amplified by PCR using BBa_K1694035 as a template with a primer containing BamHI site and digested by EcoRI and SpeI, followed by cloning onto pSB1C3 which was cut by EcoRI and SpeI. The part has been confirmed by sequencing. false Chen, Pei-En annotation2521542 1 Lpp range2521542 1 1 87 annotation2521543 1 linker range2521543 1 88 90 annotation2521545 1 BamHI range2521545 1 436 441 annotation2521544 1 OmpA range2521544 1 91 435 BBa_J23101 1 BBa_J23101 constitutive promoter family member 2006-08-03T11:00:00Z 2015-08-31T04:08:40Z later Released HQ 2013 later false true _52_ 0 483 95 In stock true N/A true John Anderson BBa_B0034 1 BBa_B0034 RBS (Elowitz 1999) -- defines RBS efficiency 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 RBS based on Elowitz repressilator. false true _1_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. <p>No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: <a href="mailto:(bchow@media.mit.edu)">Brian Chow</a> true Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003. annotation23325 1 conserved range23325 1 5 8 BBa_B0034_sequence 1 aaagaggagaaa BBa_K1991004_sequence 1 atgaaagcaaccaagctggttctgggtgccgtgattctgggcagtaccctgttagcaggttgttctagcaatgccaaaatcgaccaaggcatcaacaacaatggcccgacccacgaaaaccagctgggtgccggtgcctttggtggttatcaggtgaacccgtacgtgggctttgaaatgggctatgattggctgggccgcatgccgtacaaaggcagtgtggagaacggcgcctataaagcacagggcgtgcagctgacagcaaaactgggctaccctattaccgacgacctggacatctacacacgcttaggcggcatggtgtggcgcgccgataccaagagcaacgtgtacggcaagaaccacgataccggcgtgagtccggtgtttgccggcggtgtggagtatgcaatcaccccggaaattgccacacgtggatcc BBa_J23101_sequence 1 tttacagctagctcagtcctaggtattatgctagc BBa_K1991007_sequence 1 tttacagctagctcagtcctaggtattatgctagctactagagaaagaggagaaatactagatgaaagcaaccaagctggttctgggtgccgtgattctgggcagtaccctgttagcaggttgttctagcaatgccaaaatcgaccaaggcatcaacaacaatggcccgacccacgaaaaccagctgggtgccggtgcctttggtggttatcaggtgaacccgtacgtgggctttgaaatgggctatgattggctgggccgcatgccgtacaaaggcagtgtggagaacggcgcctataaagcacagggcgtgcagctgacagcaaaactgggctaccctattaccgacgacctggacatctacacacgcttaggcggcatggtgtggcgcgccgataccaagagcaacgtgtacggcaagaaccacgataccggcgtgagtccggtgtttgccggcggtgtggagtatgcaatcaccccggaaattgccacacgtggatcc 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