BBa_J119389 1 BBa_J119389 rClone Blue: Device for GGA Cloning and Testing RBS elements and Riboswitches 2015-06-28T11:00:00Z 2015-08-31T04:08:29Z PCR of J119139 and J119384 and BbsI GGA. rClone Red allows users to clone and test new RBS elements and riboswitches without gel purification or other preparation of DNA. It is a destination vector for Golden Gate Assembly (GGA) using BsaI and ligase. A new RBS or riboswitch can be derived from synthetic oligos, PCR, or a plasmid clone. For proper assembly, the new insert must have the appropriate 4 nt sticky ends or be flanked by BsaI sites that produce the sticky ends. With reference to the top strand, the left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. BsaI always produces a 5' overhang sticky end. Successful GGA assembly replaces the reverse promoter driving GFP expression with the new RBS or riboswitch. Transcription will be initiated in the direction of the RFP coding sequence. The level of expression of RFP will depend on the efficiency of the newly cloned RBS or riboswitch. false false _613_435_ 0 606 61 Not in stock false None. false Anthony Eckdahl annotation2432911 1 GFP reverse range2432911 1 55 774 annotation2432915 1 Amil CP Blue range2432915 1 862 1530 annotation2432910 1 B0034 RBS reverse range2432910 1 781 792 annotation2432914 1 BsaI right range2432914 1 849 854 annotation2432913 1 P5 promoter range2432913 1 1 36 annotation2432916 1 P2 promoter reverse range2432916 1 803 848 annotation2432912 1 BsaI left range2432912 1 42 47 BBa_J119389_sequence 1 ttgacaattaatcatccggctcgtaatttatgtggacgactgagaccactagagttattatttgtatagttcatccatgccatgtgtaatcccagcagctgttacaaactcaagaaggaccatgtgatctctcttttcgttgggatctttcgaaagggcagattgtgtggacaggtaatggttgtctggtaaaaggacagggccatcgccaattggagtattttgttgataatggtctgctagttgaacgcttccatcttcaatgttgtgtctaattttgaagttaactttgattccattcttttgtttgtctgccatgatgtatacattgtgtgagttatagttgtattccaatttgtgtccaagaatgtttccatcttctttaaaatcaataccttttaactcgattctattaacaagggtatcaccttcaaacttgacttcagcacgtgtcttgtagttcccgtcatctttgaaaaatatagttctttcctgtacataaccttcgggcatggcactcttgaaaaagtcatgctgtttcatatgatctgggtatctcgcaaagcattgaacaccataaccgaaagtagtgacaagtgttggccatggaacaggtagttttccagtagtgcaaataaatttaagggtaagttttccgtatgttgcatcaccttcaccctctccactgacagaaaatttgtgcccattaacatcaccatctaattcaacaagaattgggacaactccagtgaaaagttcttctcctttacgcatctagtatttctcctctttaattactagatccacacattataggtacaaaaagatgcgaagtcaatactctttttggtctctgcggagatgagcgtgattgcaaagcagatgacctataaagtgtatatgagcggcaccgtgaacggccattattttgaagttgaaggtgatggtaaaggcaaaccgtatgaaggcgaacagaccgttaaactgaccgttaccaaaggtggtccgctgccgtttgcatgggatattctgagtccgcagtgtcagtatggtagcattccgtttaccaaatatccggaagatatcccggattatgtgaaacagagctttccggaaggttatacctgggaacgtattatgaactttgaagatggtgcagtttgcaccgttagcaatgatagcagcattcagggtaattgctttatctaccacgtgaaatttagcggtctgaattttccgcctaatggtccggttatgcagaaaaaaacccagggttgggaaccgaataccgaacgtctgtttgcacgtgatggcatgctgctgggtaataactttatggcactgaaactggaaggtggtggtcattatctgtgtgagtttaaaaccacctacaaagccaaaaaaccggtgaaaatgcctggttatcattatgtggatcgtaaactggatgtgaccaaccacaataaagattacaccagcgttgaacagtgcgaaattagcattgcacgtaaaccggttgttgcctaataa igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z