BBa_K2095000 1 BBa_K2095000 pLIP (SORLIPx2) 2016-10-08T11:00:00Z 2016-10-15T08:33:29Z Dunaliella sp. This basic part consists of a synthesised truncated version of the promotor of the light-inducible protein gene (LIP) in Dunaliella microalgae. Activation of the promoter has shown to be stimulated by high intensity light when introduced into Chlamydomonas Reinhardtii (1)(2). In 2013, Park S. et al. investigated the promoter of the highly inducible promoter gene (LIP) of the marine algae Dunaliella sp.(1). They discovered several light-responsive motifs by experimenting with truncations of the promotor fused with a Renilla luciferase gene introduced in C. reinhardtii. In a more recent study from 2016, they identified sequences over-represented in light-inducible promoters (SORLIP)(2). Duplication of the SORLIP sequence upstream a 100 bp truncation of the LIP promoter showed promising results in activity when stimulated by high- and medium-intensity light. This BioBrick consists of the 2xSORLIP promotor sequence followed by an intron, optimized for use C. reinhardtii. 1. Park S, Lee Y, Lee J-H, Jin E. Expression of the high light-inducible Dunaliella LIP promoter in Chlamydomonas reinhardtii. Planta [Internet]. 2013 Dec [cited 2016 Sep 4];238(6):1147???56. Available from: http://www.ncbi.nlm.nih.gov/pubmed/24043576 2. Baek K, Lee Y, Nam O, Park S, Sim SJ, Jin E. Introducing Dunaliella LIP promoter containing light-inducible motifs improves transgenic expression in Chlamydomonas reinhardtii. Biotechnol J. 2016;11(3):384???92. false false _2563_ 31143 31143 9 false The intron was added to improve gene expression in C. reinhardtii, but is not necessarily vital for its function. false Liu iGEM annotation2489053 1 SORLIPx2 range2489053 1 5 144 BBa_E1010 1 mRFP1 **highly** engineered mutant of red fluorescent protein from Discosoma striata (coral) 2004-07-27T11:00:00Z 2015-08-31T04:07:26Z Campbell et al., PNAS v99 p7877 <a href="http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=12060735">URL</a> Released HQ 2013 monomeric RFP: Red Fluorescent Protein. Excitation peak: 584 nm Emission peak: 607 nm false false _11_1_ 0 52 7 In stock false TAATAA double stop codon added (DE). Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A. true Drew Endy annotation1014044 1 mrfp1 range1014044 1 1 675 annotation2214014 1 Help:Barcodes range2214014 1 682 706 BBa_K2095002 1 BBa_K2095002 RBCS2 3 2016-10-09T11:00:00Z 2016-10-15T08:37:13Z pOpt_mVenus_Hyg expression plasmid Chlamydomonas reinhardtii RuBisCO small subunit 2 terminator obtained from the pOpt_mVenus_Hyg expression plasmid provided by Chlamydomonas Resource Center. false false _2563_ 31137 31137 9 false RBCS2 sequence was synthesized by IDT. false Liu iGEM 2016 BBa_K2095003 1 BBa_K2095003 mRFP1 regulated by pLIP 2016-10-09T11:00:00Z 2016-10-15T08:35:43Z Truncated LIP promoter (BBa_K2095000) source: Dunaliella sp, RFP (BBa_E1010) source: Discosoma striata, RBCS2 terminator (BBa_K2095002) source: pOpt_mVenus_Hyg expression plasmid. This composite part was designed by the Linkoping iGEM team 2016 to demonstrate the functionality of the light-inducible promoter, LIP, Biobrick (BBa_K2095000) A synthesised truncated version of the promotor of the light-inducible protein gene (LIP) in Dunaliella microalgae was set to control the expression of the mRFP1 reporter gene, obtained from BBa_E1010. This part is terminated by a RuBisCO terminator (BBa_K2095002). The part was created to evaluate function and potential leakage of LIP in C. reinhardtii. To provide this information, we planned to perform experiments in successfully transformed algae. This by measuring the RFP expression in transformed algae, stimulated by light of different intensities, compared to algae grown in complete darkness. Unfortunately, the transformation of this part into C. reinhardtii failed, and no results on BBa_K2095000 functionality were provided. false false _2563_ 31137 31137 9 false Used a truncated LIP promoter in order to optimize the promoters gene activating properties. false Liu iGEM 2016 component2490382 1 BBa_K2095002 component2490381 1 BBa_E1010 component2490378 1 BBa_K2095000 annotation2490378 1 BBa_K2095000 range2490378 1 1 305 annotation2490382 1 BBa_K2095002 range2490382 1 1026 1272 annotation2490381 1 BBa_E1010 range2490381 1 312 1017 BBa_E1010_sequence 1 atggcttcctccgaagacgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagacggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataacgctgatagtgctagtgtagatcgc BBa_K2095002_sequence 1 gcgcccgctccgtgtaaatggaggcgctcgttgatctgagccttgccccctgacgaacggcggtggatggaagatactgctctcaagtgctgaagcggtagcttagctccccgtttcgtgctgatcagtctttttcaacacgtaaaaagcggaggagttttgcaattttgttggttgtaacgatcctccgttgattttggcctctttctccatgggcgggctgggcgtatttgaagcggacccgcgc BBa_K2095003_sequence 1 gcgcgggctccgggccaccggagcgtgagggctccgggccacacatttgccagcctgaaaacttgccaaaaaccactcatcatcaaaacaacaaaagcttcaatcaaaactcgttcctacacccacacgaaccgacccgaacaggccaggtgagtcgacgagcaagcccggcggatcaggcagcgtgcttgcagatttgacttgcaacgcccgcattgtgtcgacgaaggcttttggctcctctgtcgctgtctcaagcagcatctaaccctgcgtcgccgtttccatttgcaggaagcttgcgctactagatggcttcctccgaagacgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagacggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataacgctgatagtgctagtgtagatcgctactagaggcgcccgctccgtgtaaatggaggcgctcgttgatctgagccttgccccctgacgaacggcggtggatggaagatactgctctcaagtgctgaagcggtagcttagctccccgtttcgtgctgatcagtctttttcaacacgtaaaaagcggaggagttttgcaattttgttggttgtaacgatcctccgttgattttggcctctttctccatgggcgggctgggcgtatttgaagcggacccgcgc BBa_K2095000_sequence 1 gcgcgggctccgggccaccggagcgtgagggctccgggccacacatttgccagcctgaaaacttgccaaaaaccactcatcatcaaaacaacaaaagcttcaatcaaaactcgttcctacacccacacgaaccgacccgaacaggccaggtgagtcgacgagcaagcccggcggatcaggcagcgtgcttgcagatttgacttgcaacgcccgcattgtgtcgacgaaggcttttggctcctctgtcgctgtctcaagcagcatctaaccctgcgtcgccgtttccatttgcaggaagcttgcgc 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