BBa_K390000 1 BBa_K390000 pSB1A3_Int (BamHI, XhoI, NdeI, NheI added) 2010-07-31T11:00:00Z 2015-05-08T01:12:19Z pSB1A3 with BamHI, XhoI, NdeI, and NheI sites mutated in outside of the terminator regions. This part is based on pSB1A3, with BamHI, XhoI, NdeI and NheI sites added through site-directed mutagenesis. This plasmid backbone retains compatibility with BioBricks (except with RFC 21 - BamBgl), its antibiotic resistance, and its high copy number in E. coli. The additional restriction sites allow the addition of DNA sequences homologous to genomic sequences in Synechocystis sp. PCC 6803 and other species, enabling the plasmid to be used to integrate the BioBrick construct and an antibiotic resistance gene into the chromosome at a specific location through homologous recombination. Integration sequences can be designed so that the BioBrick construct is added in the middle of a gene knocking the gene out in the process, or between two genes without interfering with native expression. The BioBrick construct can be assembled in E. coli cells without changes to the standard procedure, and when completely assemble, the plasmid can be introduced into the target species. Synechocystis sp. PCC 6803 naturally takes up DNA from its environment, and if sequences are homologous to regions of the genome, will naturally undergo homologous recombination. Other species may lack one or both of these abilities, and the addition of another plasmid carrying a recombinase gene may be necessary for genomic integration. false false _518_ 0 6788 9 It's complicated true BamHI, XhoI, NdeI, and NheI sites were mutated in using Stratagene's site-directed mutagenesis kit, the sites were mutated in by changing the nucleotide sequence, without the addition or removal of nucleotides. false Cody Tramp annotation2121030 1 VR range2121030 1 157 176 annotation2121038 1 BamHI Site range2121038 1 1992 1997 annotation2121026 1 suffix range2121026 1 1 21 annotation2121042 1 a > g range2121042 1 2014 2014 annotation2121037 1 AmpR (rev) range2121037 1 1025 1895 annotation2121035 1 rep(pMB1) (rev) range2121035 1 261 875 annotation2121028 1 terminator stem loop range2121028 1 30 64 annotation2121045 1 terminator stem loop range2121045 1 2107 2126 annotation2121036 1 Ori range2121036 1 276 276 annotation2121029 1 deletion of two bases range2121029 1 34 35 annotation2121043 1 VF2 range2121043 1 2017 2037 annotation2121039 1 t > a range2121039 1 1995 1995 annotation2121046 1 prefix range2121046 1 2136 2157 annotation2121027 1 E. coli his operon terminator range2121027 1 22 93 annotation2121032 1 g > a range2121032 1 219 219 annotation2121031 1 NdeI site range2121031 1 216 221 annotation2121033 1 NheI Site range2121033 1 230 235 annotation2121040 1 XhoI Site range2121040 1 2009 2014 annotation2121041 1 c > t range2121041 1 2010 2010 annotation2121044 1 end of terminator range2121044 1 2094 2097 annotation2121034 1 c > t range2121034 1 232 232 BBa_K390000_sequence 1 tactagtagcggccgctgcagtccggcaaaaaaacgggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgttatgcaggcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatatgagcaaaaggctagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggatccgcgcacatttcctcgagaagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggcagaatttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagag 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