BBa_K571005 1 BBa_K571005 R0011+acsAB/pSB1C3 2011-10-02T11:00:00Z 2015-05-08T01:12:44Z *acsAB: GenBank AB091060.1 *promoter: iGEM registry of standard biological parts The biobrick is composite element, combination of BBa_K571000 and BBa_K571003. The promoter binding to the upstream of acsAB gene, IPTG as the inducer, activates the acsAB gene, producing Cyclic-di-GMP binding domain and UDP-forming domain. The sequence of acsAB and promoter is amplified by PCR, followed by enzyme digestion and ligation to bind the 2 sequences with the backbone-pSB1C3. The order of restriction enzyme is EcoRI, XbaI and SpeI. false false _739_ 0 10422 9 It's complicated true It is a very long sequence, therefore below is the considerations to take note during cloning. *The concentration agarose gel during gel electrophoresis should be about 0.8-1% *The enzyme digestion reaction time is advised to be elongated so that digestion could be more perfect. *The respective restriction enzyme at the both end of primer should not cut the sequence of another gene false Lu-Chu Ke component2151367 1 BBa_K571000 component2151365 1 BBa_K571004 annotation2151367 1 BBa_K571000 range2151367 1 87 4739 annotation2151365 1 BBa_K571004 range2151365 1 1 80 BBa_K571000 1 BBa_K571000 acsAB/pSB1C3 2011-10-01T11:00:00Z 2015-05-08T01:12:44Z *organism: Gluconacetobacter hansenii ATCC 23769 *GenBank: AB091060.1 The acs operon in Gluconacetobacter hansenii is the mechanism that synthesize cellulose synthase, acsAB is one of the gene sequence. The acsAB will produce protein with 2 subunits; one is catalytic subunit which is able to transform UDP-glucose to UDP, another one would be the regulatory subunit of the cellulose synthase which will bind with cyclic di-GMP and activated. From the primer we design, the acsAB sequence is amplified from PCR from the genomic DNA of Gluconacetobacter hansenii, XbaI and SpeI is also added at the both end of primer as the cutting site, so that acsAB and pSB1C3 could easily bind to it. false false _739_ 0 10422 9 It's complicated false By comparative, acsAB is long base pairs gene, so we need more primer during PCR. It is also advisable to separate PCR into a few times to get the most satisfied results. false Lu-Chu Ke annotation2149953 1 acsAB range2149953 1 1 4653 BBa_K571004 1 BBa_K571004 R0011+RBS/pSB1C3 2011-10-02T11:00:00Z 2015-05-08T01:12:44Z BBa_R0011 This biobrick contains the BBa_R0011 promoter of 80 bp cloned in pSB1C3 with EcoRI and XbaI restriction site which will can be induced by IPTG. RBS is added to the end of the sequence in order to enable gene expression to perform well. false false _739_ 0 10422 9 It's complicated false none. false Lu-Chu Ke annotation2151464 1 R0011 range2151464 1 1 55 annotation2151465 1 RBS range2151465 1 64 80 BBa_K571005_sequence 1 aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcacatactagagttaaggaggtaaaaaaatactagatgccagaggttcggtcgtcaacgcagtcagagtcaggaatgtcacagtggatggggaaaattctctccattcgcggtgctgggctgattattggtgtttttggcctgtgtgcgctgattgcggctacgtccgtgaccctgccgccagaacagcagttgattgtggcatttgtatgtgtcgtgatcttttttattgtcggtcataagcccagccgtcggtcccagattttccttgaagtgctgtcagggctggtttcgctgcgctatctgacatggcgcctgacggaaacgctttcattcgatacatggttgcagggtctgcttgggacaatgcttctggtggcggaactttacgccctgatgatgctgttcctcagctacttccagacgatcgcgccgctgcatcgtgcgcctctgccgctgccgccgaaccctgacgaatggcccacggtcgatatcttcgtcccgacctataacgaagaactgagcattgtccgcctgacggtgctgggatcactggggattgactggccaccggaaaaggtgcgggttcatatccttgatgacggccgtcgtcctgaattcgccgcctttgccgctgaatgtggcgcgaattatatcgcccgcccgacgaacgaacatgcaaaggccggtaatcttaactatgccattggtcataccgatggtgattacatcctgatctttgactgcgaccacgtcccgacccgcgccttcctgcagttgacaatgggctggatggtcgaagacccgaagatcgcgctgatgcagaccccgcatcacttctattcccccgacccgttccagcggaacctgtcggctggttatcgtaccccgcccgagggcaacctgttttatggcgtggtgcaggatggcaacgatttctgggatgcgaccttcttttgcgggtcatgtgcaatcctgcgtcgcacggcgattgagcagatcggcggctttgcgacccagaccgtgaccgaagacgcgcataccgcactcaagatgcagcgtctgggctggtccacggcctatctgcgtatcccgcttgccggtggtctcgcgacggaacgcctgatcctgcatatcggacagcgcgtgcgctgggcgcgtgggatgctgcagatcttccgcatcgacaatcctctgttcgggcgtggcctgtcatgggggcagcggctttgttacctgtcggccatgacgtcgttcctgttcgctgtcccgcgcgtcatcttcctgagctccccgctggcgttcctgttctttgggcagaacatcattgccgcgtcgccgctcgcgctgctggcctatgccatcccgcatatgttccacgccgtcggcacggcgtcgaagatcaacaagggctggcgctactccttctggagtgaggtctatgaaaccaccatggcgctgttcctggtgcgcgtgacgattgtcaccctgctcagcccttcacgtgggaagttcaacgtgacggacaagggcgggttgcttgaaaaaggttatttcgaccttggcgccgtctatccgaacatcatccttggcctgatcatgttcggcggtctggcgcgtggtgtctatgaactgtctttcggccatctcgaccagatcgccgaacgtgcctacctgctgaactccgcttgggcgatgctcagcctcatcatcatccttgcggccatcgccgtggggcgtgaaacacagcagaaacgcaacagtcatcgcatccccgcaaccatcccggtggaagtggcgaatgccgatgggtccatcatcgtgacgggcgtgaccgaggacctgtccatgggtggggccgcggtaaagatgtcatggcctgcgaagctgtcggggccgacgccggtttatatccgtactgtccttgatggagaggaactgatcctgcccgccaggatcatccgtgctggcaacgggcgggggatcttcatctggacgattgataacctgcagcaggaattctcggttatccgcctggtgttcggccgtgccgacgcatgggttgactggggcaattacaaggccgaccgcccgctgctcagcctcatggacatggttctcagcgtcaagggcctgttccgttcaagcggcgacatcgtccatcgcagttccccaaccaagccttcggctggcaatgccctgtctgacgatacgaacaacccgtcacgcaaggagcgtgtgctgaagggaaccgtgaaaatggtttcgcttctggcgctgctgacatttgcttcctcggcacaggcggcgtcagcgcccagggccgtcgcggcgaaggccccggcccatcagcccgaagcctctgacctgccgccgttgcccgcgctcctgccggccaccagcggtgcggcgcaggcgggttcgggcgatgccggcgccgatggacccggcagccccacgggccagcccctggcggccgacagtgccgatgcgctggtggaaaatgcggaaaatacgtccgatacggcgactgtccataattataccctcaaggatcttggcgccgcagggtccatcacaatgcgtggccttgccccgttgcaggggatcgagttcgggattccctctgaccagcttgtgacgtccgcgcgccttgtgctgtcgggttcgatgtcgcccaacctgcgtccggaaaccaattcggtcacaatgacgctgaacgagcagtatatcggcacgttgcgtcccgacccggcgcacccgacattcggccccatgtcgttcgagatcaacccgatcttcttcgtcagcggaaaccgtctgaacttcaacttcgcctccgggtcgaagggatgttcggacatcacgaacgatacgctgtgggccacgatctcgcagaactcgcagcttcagatcacgacaatcgcgctgccgccgcgtcgcctgctgtcacgtctgccgcagcccttctatgacaagaacgtgcgccagcatgttacggtcccgatggtgctggcgcagacctatgacccgcagatactcaagtccgcggggattctcgcttcgtggtttggcaagcagacggacttcctcggcgtgacgttcccggtgtcgtccaccatcccacagagtggcaacgccatcctgatcggcgtggccgatgaactgccgaccagcttcgggcggccgcaggtcaatggcccggcggttctggaactgccgaacccgtcggatgcaaatgccacgatcctggtggtgacggggcgtgaccgtgatgaggttattaccgcgagcaagggcatcgccttcgcgtctgctcccctgccgaccgacagccatatggatgtcgcgccggtcgatatcgccccgcgcaagcccaatgacgcgccatcttttatcgcgatggaccatccggtgcgctttggcgaccttgtaacggcaagcaagctacagggaaccggctttacgtccggtgtgctgtcggttccgttccgcattccgcccgacctttatacgtggcgtaaccgcccgtacaagatgcaggtacgtttccgttcccccgcgggggaggcgaaggatgtcgagaagtcacgtctcgatgtcgggatcaacgaggtttacctgcattcctatccgctgcgcgaaacgcatggcctgattggcgcggttttgcagggtgtcggccttgcccgccctgcaagcggcatgcaggtgcatgatctcgacgtaccgccgtggaccgtgttcgggcaggatcagttgaacttctactttgacgcgatgccccttgcgcgcggaatctgtcaaagtggcgcggcaaacaatgcattccatcttgggcttgacccggattccaccattgatttttcccgtgcccatcacatcgcccagatgcccaaccttgcctatatggcgacggtcggttttcctttcaccacctatgccgatctgtcgcagacggcggttgttctgcctgaacacccgaatgccgcgactgttggcgcctatcttgacctgatggggttcatgggcgcggcgacgtggtatccggttgcgggcgtggacattgtgtcggccgatcatgtcagtgacgttgcggaccgtaacctgctggtgatttcgacgctggcgacaagtggcgagatcgcgccgctgctgtcacgttcatcctacgaagtggcggatggtcatttgcgcacggtgtcgcatgcgtccgcgctggataacgcgatcaaggcggtcgacgatccgctgacggctttccgcgaccgtgacagcaagccgcaggatgtcgatacccccctgacgggtggtgtcggcgcgatgatcgaggcggaatcccccctgacggcgggccgcaccgttctggcgctgctgtcgtctgacggtgcggggctgaacaacctccttcagatgttgggggagcgcaagaagcaggcgaatatccagggggacctggttgttgcccatggcgaggacctgtcttcgtaccgcacttcgccggtctatacgatcggcaccctgccgctctggctgtggccggactggtatatgcacaacagaccggttcgtgtgctgcttgtcggcctgttgggatgtattctgatcgtcagtgttcttgcacgggctctggcgcggcatgcggcccggcgtttcaagcagcttgaggatgagaggcgcaagtcgtga BBa_K571004_sequence 1 aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcacatactagagttaaggaggtaaaaaaa BBa_K571000_sequence 1 atgccagaggttcggtcgtcaacgcagtcagagtcaggaatgtcacagtggatggggaaaattctctccattcgcggtgctgggctgattattggtgtttttggcctgtgtgcgctgattgcggctacgtccgtgaccctgccgccagaacagcagttgattgtggcatttgtatgtgtcgtgatcttttttattgtcggtcataagcccagccgtcggtcccagattttccttgaagtgctgtcagggctggtttcgctgcgctatctgacatggcgcctgacggaaacgctttcattcgatacatggttgcagggtctgcttgggacaatgcttctggtggcggaactttacgccctgatgatgctgttcctcagctacttccagacgatcgcgccgctgcatcgtgcgcctctgccgctgccgccgaaccctgacgaatggcccacggtcgatatcttcgtcccgacctataacgaagaactgagcattgtccgcctgacggtgctgggatcactggggattgactggccaccggaaaaggtgcgggttcatatccttgatgacggccgtcgtcctgaattcgccgcctttgccgctgaatgtggcgcgaattatatcgcccgcccgacgaacgaacatgcaaaggccggtaatcttaactatgccattggtcataccgatggtgattacatcctgatctttgactgcgaccacgtcccgacccgcgccttcctgcagttgacaatgggctggatggtcgaagacccgaagatcgcgctgatgcagaccccgcatcacttctattcccccgacccgttccagcggaacctgtcggctggttatcgtaccccgcccgagggcaacctgttttatggcgtggtgcaggatggcaacgatttctgggatgcgaccttcttttgcgggtcatgtgcaatcctgcgtcgcacggcgattgagcagatcggcggctttgcgacccagaccgtgaccgaagacgcgcataccgcactcaagatgcagcgtctgggctggtccacggcctatctgcgtatcccgcttgccggtggtctcgcgacggaacgcctgatcctgcatatcggacagcgcgtgcgctgggcgcgtgggatgctgcagatcttccgcatcgacaatcctctgttcgggcgtggcctgtcatgggggcagcggctttgttacctgtcggccatgacgtcgttcctgttcgctgtcccgcgcgtcatcttcctgagctccccgctggcgttcctgttctttgggcagaacatcattgccgcgtcgccgctcgcgctgctggcctatgccatcccgcatatgttccacgccgtcggcacggcgtcgaagatcaacaagggctggcgctactccttctggagtgaggtctatgaaaccaccatggcgctgttcctggtgcgcgtgacgattgtcaccctgctcagcccttcacgtgggaagttcaacgtgacggacaagggcgggttgcttgaaaaaggttatttcgaccttggcgccgtctatccgaacatcatccttggcctgatcatgttcggcggtctggcgcgtggtgtctatgaactgtctttcggccatctcgaccagatcgccgaacgtgcctacctgctgaactccgcttgggcgatgctcagcctcatcatcatccttgcggccatcgccgtggggcgtgaaacacagcagaaacgcaacagtcatcgcatccccgcaaccatcccggtggaagtggcgaatgccgatgggtccatcatcgtgacgggcgtgaccgaggacctgtccatgggtggggccgcggtaaagatgtcatggcctgcgaagctgtcggggccgacgccggtttatatccgtactgtccttgatggagaggaactgatcctgcccgccaggatcatccgtgctggcaacgggcgggggatcttcatctggacgattgataacctgcagcaggaattctcggttatccgcctggtgttcggccgtgccgacgcatgggttgactggggcaattacaaggccgaccgcccgctgctcagcctcatggacatggttctcagcgtcaagggcctgttccgttcaagcggcgacatcgtccatcgcagttccccaaccaagccttcggctggcaatgccctgtctgacgatacgaacaacccgtcacgcaaggagcgtgtgctgaagggaaccgtgaaaatggtttcgcttctggcgctgctgacatttgcttcctcggcacaggcggcgtcagcgcccagggccgtcgcggcgaaggccccggcccatcagcccgaagcctctgacctgccgccgttgcccgcgctcctgccggccaccagcggtgcggcgcaggcgggttcgggcgatgccggcgccgatggacccggcagccccacgggccagcccctggcggccgacagtgccgatgcgctggtggaaaatgcggaaaatacgtccgatacggcgactgtccataattataccctcaaggatcttggcgccgcagggtccatcacaatgcgtggccttgccccgttgcaggggatcgagttcgggattccctctgaccagcttgtgacgtccgcgcgccttgtgctgtcgggttcgatgtcgcccaacctgcgtccggaaaccaattcggtcacaatgacgctgaacgagcagtatatcggcacgttgcgtcccgacccggcgcacccgacattcggccccatgtcgttcgagatcaacccgatcttcttcgtcagcggaaaccgtctgaacttcaacttcgcctccgggtcgaagggatgttcggacatcacgaacgatacgctgtgggccacgatctcgcagaactcgcagcttcagatcacgacaatcgcgctgccgccgcgtcgcctgctgtcacgtctgccgcagcccttctatgacaagaacgtgcgccagcatgttacggtcccgatggtgctggcgcagacctatgacccgcagatactcaagtccgcggggattctcgcttcgtggtttggcaagcagacggacttcctcggcgtgacgttcccggtgtcgtccaccatcccacagagtggcaacgccatcctgatcggcgtggccgatgaactgccgaccagcttcgggcggccgcaggtcaatggcccggcggttctggaactgccgaacccgtcggatgcaaatgccacgatcctggtggtgacggggcgtgaccgtgatgaggttattaccgcgagcaagggcatcgccttcgcgtctgctcccctgccgaccgacagccatatggatgtcgcgccggtcgatatcgccccgcgcaagcccaatgacgcgccatcttttatcgcgatggaccatccggtgcgctttggcgaccttgtaacggcaagcaagctacagggaaccggctttacgtccggtgtgctgtcggttccgttccgcattccgcccgacctttatacgtggcgtaaccgcccgtacaagatgcaggtacgtttccgttcccccgcgggggaggcgaaggatgtcgagaagtcacgtctcgatgtcgggatcaacgaggtttacctgcattcctatccgctgcgcgaaacgcatggcctgattggcgcggttttgcagggtgtcggccttgcccgccctgcaagcggcatgcaggtgcatgatctcgacgtaccgccgtggaccgtgttcgggcaggatcagttgaacttctactttgacgcgatgccccttgcgcgcggaatctgtcaaagtggcgcggcaaacaatgcattccatcttgggcttgacccggattccaccattgatttttcccgtgcccatcacatcgcccagatgcccaaccttgcctatatggcgacggtcggttttcctttcaccacctatgccgatctgtcgcagacggcggttgttctgcctgaacacccgaatgccgcgactgttggcgcctatcttgacctgatggggttcatgggcgcggcgacgtggtatccggttgcgggcgtggacattgtgtcggccgatcatgtcagtgacgttgcggaccgtaacctgctggtgatttcgacgctggcgacaagtggcgagatcgcgccgctgctgtcacgttcatcctacgaagtggcggatggtcatttgcgcacggtgtcgcatgcgtccgcgctggataacgcgatcaaggcggtcgacgatccgctgacggctttccgcgaccgtgacagcaagccgcaggatgtcgatacccccctgacgggtggtgtcggcgcgatgatcgaggcggaatcccccctgacggcgggccgcaccgttctggcgctgctgtcgtctgacggtgcggggctgaacaacctccttcagatgttgggggagcgcaagaagcaggcgaatatccagggggacctggttgttgcccatggcgaggacctgtcttcgtaccgcacttcgccggtctatacgatcggcaccctgccgctctggctgtggccggactggtatatgcacaacagaccggttcgtgtgctgcttgtcggcctgttgggatgtattctgatcgtcagtgttcttgcacgggctctggcgcggcatgcggcccggcgtttcaagcagcttgaggatgagaggcgcaagtcgtga 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