BBa_J119127 1 BBa_J119127 For Testing New Promoters via Golden Gate Assembly with BsmBI 2012-10-29T12:00:00Z 2015-08-31T04:08:27Z Golden Gate assembled of PCR product and BBa_J100091 using BsaI and Ligase. This part allows users to clone and test new promoters without gel purification or other preparation of DNA. It is a destination vector for [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate Golden Gate Assembly] (GGA) using BsmBI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsmBI sites that produce the 4 nt overhangs required for assembly (see [[Part:BBa_J119022]] as an example, or the picture below). Successful GGA assembly replaces the double terminator in '''BBa_J119127''' with the new promoter. Upon assembly, a functional new promoter should cause RFP expression. BBa_J119127 incorporates the BD18 bicistronic translational junction (see [[Part:BBa_J119024]]) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org. [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate_Assembly_protocol See the full GGA protocol online.]<br><center> [[Image:J100028.png]] false false _613_120_ 0 606 61 Not in stock false None. false Todd Eckdahl annotation2214062 1 BsmBI sticky end right range2214062 1 114 117 annotation2214063 1 BD18 biscistron range2214063 1 118 205 annotation2214069 1 Double stop for E1010 range2214069 1 878 883 annotation2214065 1 RBS for RFP range2214065 1 188 196 annotation2214059 1 BsmBI cuts left range2214059 1 6 11 annotation2214067 1 RFP E1010 range2214067 1 203 883 annotation2214066 1 Stop for BD18 leader range2214066 1 201 203 annotation2214061 1 BsmBI cuts right range2214061 1 107 112 annotation2214068 1 Start for RFP range2214068 1 203 205 annotation2214064 1 leader RBS range2214064 1 135 143 annotation2214060 1 TT B0014 range2214060 1 12 106 annotation2214058 1 BsmBI sticky end left range2214058 1 1 4 BBa_J119135 1 BBa_J119135 Backwards RBS (B0030)-GFP (E0040) 2013-01-10T12:00:00Z 2015-08-31T04:08:27Z PCR amplification of E5500. This composite part is a backwards version of E5500, which contains RBS (B0030)-GFP (E0040). It can be expressed by backwards promoters. false false _613_435_ 0 606 61 Not in stock false None. false Claire Shinneman, Paul Frederick annotation2213819 1 GFP reverse range2213819 1 1 719 annotation2213818 1 RBS reverse range2213818 1 727 741 BBa_J119136 1 BBa_J119136 Device for Testing New Promoters via Golden Gate Assembly 2013-01-10T12:00:00Z 2015-08-31T04:08:27Z PCR product from amplification of E5500 ws cloned into BBa_J119127. This part will allow users to clone and test new promoters without gel purification or other preparation of DNA - it is ideal for teaching labs. It is a destination vector for Golden Gate Assembly (GGA) using BsmBI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsmBI sites that produce the 4 nt overhangs required for assembly. The left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. Successful GGA assembly replaces the double terminator in with the new promoter. Upon assembly, a functional new promoter with activity to the right should cause RFP expression. A promoter with activity to the left will cause GFP expression. The part incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org. false false _613_435_ 0 606 61 Not in stock false None. false Claire Shinneman, Paul Frederick component2214085 1 BBa_J119127 component2214072 1 BBa_J119135 annotation2214072 1 BBa_J119135 range2214072 1 1 741 annotation2214085 1 BBa_J119127 range2214085 1 750 1632 BBa_J119135_sequence 1 ttattatttgtatagttcatccatgccatgtgtaatcccagcagctgttacaaactcaagaaggaccatgtggtctctcttttcgttgggatctttcgaaagggcagattgtgtggacaggtaatggttgtctggtaaaaggacagggccatcgccaattggagtattttgttgataatggtctgctagttgaacgcttccatcttcaatgttgtgtctaattttgaagttaactttgattccattcttttgtttgtctgccatgatgtatacattgtgtgagttatagttgtattccaatttgtgtccaagaatgtttccatcttctttaaaatcaataccttttaactcgattctattaacaagggtatcaccttcaaacttgacttcagcacgtgtcttgtagttcccgtcatctttgaaaaatatagttctttcctgtacataaccttcgggcatggcactcttgaaaaagtcatgctgtttcatatgatctgggtatctcgcaaagcattgaacaccataaccgaaagtagtgacaagtgttggccatggaacaggtagttttccagtagtgcaaataaatttaagggtaagttttccgtatgttgcatcaccttcaccctctccactgacagaaaatttgtgcccattaacatcaccatctaattcaacaagaattgggacaactccagtgaaaagttcttctcctttacgcatctagtatttctcctctttaat BBa_J119127_sequence 1 cgactgagacgtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattatttcgtctcagcgggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcgacggagcgtttctaatggcttcctccgaagatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagattacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataa BBa_J119136_sequence 1 ttattatttgtatagttcatccatgccatgtgtaatcccagcagctgttacaaactcaagaaggaccatgtggtctctcttttcgttgggatctttcgaaagggcagattgtgtggacaggtaatggttgtctggtaaaaggacagggccatcgccaattggagtattttgttgataatggtctgctagttgaacgcttccatcttcaatgttgtgtctaattttgaagttaactttgattccattcttttgtttgtctgccatgatgtatacattgtgtgagttatagttgtattccaatttgtgtccaagaatgtttccatcttctttaaaatcaataccttttaactcgattctattaacaagggtatcaccttcaaacttgacttcagcacgtgtcttgtagttcccgtcatctttgaaaaatatagttctttcctgtacataaccttcgggcatggcactcttgaaaaagtcatgctgtttcatatgatctgggtatctcgcaaagcattgaacaccataaccgaaagtagtgacaagtgttggccatggaacaggtagttttccagtagtgcaaataaatttaagggtaagttttccgtatgttgcatcaccttcaccctctccactgacagaaaatttgtgcccattaacatcaccatctaattcaacaagaattgggacaactccagtgaaaagttcttctcctttacgcatctagtatttctcctctttaattactagagcgactgagacgtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattatttcgtctcagcgggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcgacggagcgtttctaatggcttcctccgaagatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagattacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataa 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