BBa_J100307 1 BBa_J100307 Variant of repClone Red (J100205) 2016-10-07T11:00:00Z 2016-10-10T11:42:44Z The two main indicator proteins are GFP (BBa_I746916) and RFP (BBa_E1010). There are some SNPs in the GFP sequence due to codon optimization; these have been verified by sequencing and their functionality has been verified experimentally. This part is a variant of repClone Red (J100205), with some differences in the TetR and GFP. We designed repClone Red to test the function of repressors. Internal BsaI sites allow the insertion of the Ptet promoter and removal of the backwards P2 promoter pointed towards GFP. Successful cloning of the Ptet promoter will change colonies from green to not green. Variations on the Ptet promoter, or introduction of anhydrotetrocycline (aTc), will affect the activity of the repressor, as indicated by the fluorescence of colonies containing the plasmid. This can be used in synthetic biology laboratories or for undergraduate introductory biology lab modules. Variations from the registry sequence for TetR and GFP were sequence verified, and functionality was experimentally verified. false false _578_ 18858 18858 9 false The stop codons of the RBS and start codons of the proteins overlap to increase effective translation. This is cloned into pUC-IDT-Amp (BsaI site removed) with modified Bba prefix so that there is only an EcoRI site upstream and a normal Bba suffix. false Monica Prudencio annotation2488213 1 BsaI Recognition Site range2488213 1 1517 1522 annotation2488211 1 T->C range2488211 1 953 953 annotation2488207 1 BD24 range2488207 1 640 727 annotation2488214 1 P2 Promoter range2488214 1 1523 1568 annotation2488210 1 T->C range2488210 1 945 945 annotation2488218 1 RFP range2488218 1 1734 2414 annotation2488212 1 B0034 range2488212 1 1490 1501 annotation2488215 1 Terminator L3S2P21 range2488215 1 1577 1637 annotation2488209 1 GFP Optimized Sequence range2488209 1 764 1483 annotation2488216 1 BsaI Recognition Site range2488216 1 1638 1643 annotation2488206 1 T->G range2488206 1 243 243 annotation2488205 1 tetR Repressor C0041 range2488205 1 1 639 annotation2488208 1 P5 Promoter range2488208 1 728 763 annotation2488217 1 BD18 range2488217 1 1649 1736 BBa_J100307_sequence 1 cggccgctactagtattattagctaccgctttcacatttcagttgtttttccagtccgcaaataatcagttctaaaccaaacagaaaggcaggttctgcaccctgatgatcaaacagttcaatggcctgacgcagcagcggaggcatgctatcggtggtcggggtttcgcgttcttcttttgcaacctgatgttcttgatcttccagaacacaacccagggtaaaatgaccaactgcgctcagtgcatacagtgcgttttccagactaaaaccttgctgacacagaaatgccagctgattttccagggtttcatactgtttttcggtcggacgggtgcccagatgaacttttgcaccatcacgatggctcagcagggcacaacgaaagctttttgcattattacgcagaaaatcctgccagctttcaccttccagcggacaaaaatgggtatgatgacgatccagcatttcaattgccagtgcatccagcagtgcacgtttgtttttaacatgccaatacagggtcggctgttcaacacccagtttctgtgccagtttacgggtggtcagaccttcaataccaacttcattcagcagttccagtgcgctattaatcactttgcttttatccagacggctcattagaaaccgtccatcgcatgattaagatgtttcagtacgaaaattgctttcattgttgatctcctttttaagtgaacttgggccctccacataaattacgagccggatgattaattgtcaattattatttgtatagttcatccatgccatgtgtaatcccagcagctgttacaaactcaagaaggaccatgtgatctctctcttcgttgggatctttcgaaagggcagattgtgtggacaggtaatggttgtctggtaaaaggacagggccatcgccaattggagtattttgttgataatggcctgctagctgaacgcttccatcttcaatgttgtgtctaattttgaagttaactttgattccattcttttgtttgtctgccatgatgtatacattgtgtgagttatagttgtattccaatttgtgtccaagaatatttccatcttctttaaaatcaataccttttaactcgattctattaacaagggtatcaccttcaaacttgacttcagcacgtgtcttgtagttcccgtcatctttgaaaaatacagttctttcctgtacataaccttcgggcatggcactcttgaaaaagtcatgctgtttcatatgatctgggtatctcgcaaagcattgaacaccataaccgaaagtagtgacaagtgttggccatggaacaggtagttttccagtagtgcaaataaatttaagggtaagttttccgtatgttgcatcaccttcaccctctccactgacagaaaatttgtgcccattaacatcgccatctaattcaacaagaattgggacaactccagtgaaaagttcttctcctttacgcacctagtatttctcctctttaattactagacgacagagacctccacacattataggtacaaaaagatgcgaagtcaatactctttttggccaattctcggtaccaaattccagaaaagaggcctcccgaaaggggggccttttttcgttttggtccggtctctgcgggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcgacggagcgtttctaatggcttcctccgaagatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagattacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataa 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