LOCUS Exported 4139 bp DNA circular SYN 24-JUL-2020 DEFINITION synthetic linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2001) AUTHORS Keoni Gandall TITLE Direct Submission JOURNAL Exported from Poly. FEATURES Location/Qualifiers terminator 1..4139 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2176..2223 /note="transcription terminator for bacteriophage T7 RNA polymerase" /label="T7 terminator" primer_bind 4071..4087 /label="M13 for" /note="common sequencing primer, one of multiple similar variants" terminator 4088..4119 /label="tonB terminator" /note="bidirectional E. coli tonB-P14 transcription terminator" primer_bind 1..20 /label="dialout_for" /note="Reverse dialout primer. Unique pair for each FreeGenes part." primer_bind 2059..2078 /label="dialout_rev" /note="Forward dialout primer. Unique pair for each FreeGenes part." CDS 29..2044 /label="E. coli DNA Ligase" /note="" /translation="MESIEQQLTELRTTLRHHEYLYHVMDAPEIPDAEYDRLMRELRELETKHPELITPDSPTQRVGAAPLAAFSQIRHEVPMLSLDNVFDEESFLAFNKRVQDRLKNNEKVTWCCELKLDGLAVSILYENGVLVSAATRGDGTTGEDITSNVRTIRAIPLKLHGENIPARLEVRGEVFLPQAGFEKINEDARRTGGKVFANPRNAAAGSLRQLDPRITAKRPLTFFCYGVGVLEGGELPDTHLGRLLQFKKWGLPVSDRVTLCESAEEVLAFYHKVEEDRPTLGFDIDGVVIKVNSLAQQEQLGFVARAPRWAVAFKFPAQEQMTFVRDVEFQVGRTGAITPVARLEPVHVAGVLVSNATLHNADEIERLGLRIGDKVVIRRAGDVIPQVVNVVLSERPEDTREVVFPTHCPVCGSDVERVEGEAVARCTGGLICGAQRKESLKHFVSRRAMDVDGMGDKIIDQLVEKEYVHTPADLFKLTAGKLTGLERMGPKSAQNVVNALEKAKETTFARFLYALGIREVGEATAAGLAAYFGTLEALEAASIEELQKVPDVGIVVASHVHNFFAEESNRNVISELLAEGVHWPAPIVINAEEIDSPFAGKTVVLTGSLSQMSRDDAKARLVELGAKVAGSVSKKTDLVIAGEAAGSKLAKAQELGIEVIDEAEMLRLLGS*" primer_bind complement(2228..2244) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" rep_origin complement(2311..2899) /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3090..3950) /label="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*" promoter complement(3951..4055) /label="bla promoter" /note="ampicillin resistance gene promoter" misc_feature 2079..2138 /label="FreeGenes Barcode" /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." ORIGIN 1 AAAGGGGCCG TCAATATCAG GGTCTCTAAT GGAATCCATC GAACAGCAGC TGACTGAACT 61 GCGCACCACC CTGCGTCATC ATGAATACCT GTACCATGTC ATGGATGCGC CGGAAATCCC 121 GGATGCGGAA TACGATCGCC TGATGCGTGA GCTCCGTGAA CTGGAAACGA AACACCCGGA 181 ACTGATTACC CCGGACTCTC CAACCCAGCG TGTGGGTGCC GCTCCGCTGG CTGCTTTTTC 241 TCAGATCCGT CACGAAGTTC CGATGTTAAG CCTGGACAAC GTTTTTGACG AAGAATCATT 301 CTTGGCCTTC AACAAACGTG TGCAGGATCG TCTGAAAAAC AACGAAAAAG TTACTTGGTG 361 TTGCGAGCTC AAGCTTGATG GCCTGGCGGT TAGCATCCTG TACGAAAACG GTGTTCTGGT 421 TTCTGCCGCA ACCCGCGGTG ATGGTACCAC CGGTGAAGAT ATCACTAGCA ACGTGCGTAC 481 CATTCGTGCA ATCCCGCTGA AACTGCACGG AGAAAACATC CCGGCGCGTT TAGAAGTCCG 541 TGGTGAGGTA TTCCTGCCGC AGGCCGGCTT CGAAAAGATC AACGAGGACG CCCGTCGTAC 601 TGGGGGCAAA GTGTTCGCCA ACCCGCGCAA CGCTGCGGCT GGCAGCCTGC GTCAGTTGGA 661 CCCGCGTATC ACCGCAAAAC GTCCGCTGAC CTTTTTCTGC TACGGTGTTG GCGTTCTGGA 721 AGGTGGCGAA CTGCCGGATA CCCACCTGGG TCGCCTGCTG CAATTCAAAA AATGGGGCCT 781 GCCGGTGAGC GACCGTGTTA CTCTCTGTGA GTCCGCAGAG GAGGTTCTGG CCTTCTACCA 841 TAAGGTTGAA GAAGATAGAC CGACCCTGGG TTTCGACATT GACGGCGTAG TGATCAAAGT 901 TAACTCCCTG GCACAGCAGG AACAGCTGGG TTTCGTTGCG CGTGCACCGC GTTGGGCAGT 961 GGCTTTCAAA TTCCCCGCAC AAGAACAGAT GACTTTTGTT CGTGACGTAG AGTTCCAGGT 1021 TGGTCGCACC GGCGCTATTA CCCCAGTTGC TCGCCTGGAA CCGGTGCACG TTGCTGGCGT 1081 TTTGGTATCC AACGCAACTC TGCATAACGC TGATGAGATT GAGCGCTTGG GTCTGCGTAT 1141 CGGCGATAAG GTTGTGATCC GCCGCGCTGG TGATGTTATT CCGCAGGTAG TAAACGTGGT 1201 TCTGTCTGAA CGTCCTGAAG ATACCCGCGA AGTGGTATTC CCGACGCACT GCCCGGTCTG 1261 CGGTAGTGAC GTTGAACGTG TGGAGGGTGA AGCTGTGGCA CGTTGTACCG GCGGCCTGAT 1321 CTGTGGCGCC CAGCGTAAAG AGAGCTTAAA ACACTTTGTT TCCCGTCGTG CAATGGATGT 1381 TGACGGTATG GGTGATAAAA TCATCGACCA GCTGGTGGAA AAAGAATACG TTCACACGCC 1441 GGCCGACCTG TTCAAACTGA CCGCAGGGAA GCTGACCGGT CTGGAACGTA TGGGCCCGAA 1501 ATCCGCTCAA AATGTTGTTA ACGCACTGGA GAAGGCAAAA GAAACGACCT TTGCGCGTTT 1561 TCTGTACGCG CTGGGTATCC GCGAAGTTGG TGAGGCAACC GCGGCAGGCC TGGCAGCTTA 1621 CTTCGGAACT TTAGAAGCGC TGGAAGCGGC GTCCATCGAA GAACTGCAAA AAGTCCCGGA 1681 CGTAGGCATT GTTGTAGCGT CGCACGTTCA CAACTTCTTC GCTGAAGAAT CTAACCGTAA 1741 CGTTATCTCC GAGCTGCTTG CGGAGGGCGT ACACTGGCCA GCGCCGATCG TCATCAACGC 1801 GGAAGAGATC GATTCTCCTT TCGCCGGTAA GACCGTCGTG CTGACCGGTA GCCTCAGCCA 1861 GATGTCCCGT GACGATGCGA AGGCGCGCCT GGTAGAACTG GGTGCGAAAG TTGCCGGCTC 1921 CGTTTCTAAG AAAACGGATC TGGTGATCGC TGGTGAAGCG GCGGGTTCTA AATTGGCTAA 1981 AGCACAGGAG CTGGGCATCG AAGTAATTGA TGAAGCAGAA ATGCTGCGTC TGCTGGGTTC 2041 CTGAAGAGCT TTGAGACCAG ATAGTCACGC ACAAGACCAA CACGCTTCCC TGTCGGTTGT 2101 AGGGATAACA GGGTAATAAC GGGATCAACC GCGTCTGCCG CTGAGGTGTC AATCGTCGGA 2161 GCCGCTGAGC AATAACTAGC ATAACCCCTT GGGGCCTCTA AACGGGTCTT GAGGGGTTTT 2221 TTGCATGGTC ATAGCTGTTT CCTGAGAGCT TGGCAGGTGA TGACACACAT TAACAAATTT 2281 CGTGAGGAGT CTCCAGAAGA ATGCCATTAA TTTCCATAGG CTCCGCCCCC CTGACGAGCA 2341 TCACAAAAAT CGACGCTCAA GTCAGAGGTG GCGAAACCCG ACAGGACTAT AAAGATACCA 2401 GGCGTTTCCC CCTGGAAGCT CCCTCGTGCG CTCTCCTGTT CCGACCCTGC CGCTTACCGG 2461 ATACCTGTCC GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT TCTCATAGCT CACGCTGTAG 2521 GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC TGTGTGCACG AACCCCCCGT 2581 TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT GAGTCCAACC CGGTAAGACA 2641 CGACTTATCG CCACTGGCAG CAGCCACTGG TAACAGGATT AGCAGAGCGA GGTATGTAGG 2701 CGGTGCTACA GAGTTCTTGA AGTGGTGGCC TAACTACGGC TACACTAGAA GAACAGTATT 2761 TGGTATCTGC GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA GCTCTTGATC 2821 CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC AGATTACGCG 2881 CAGAAAAAAA GGATCTCAAG AAGGCCTACT ATTAGCAACA ACGATCCTTT GATCTTTTCT 2941 ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT CATGAGATTA 3001 TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT GAAGTTTTAA ATCAATCTAA 3061 AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT TAATCAGTGA GGCACCTATC 3121 TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC TCCCCGTCGT GTAGATAACT 3181 ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA TGATACCGCG AGAACCACGC 3241 TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG GAAGGGCCGA GCGCAGAAGT 3301 GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT GTTGCCGGGA AGCTAGAGTA 3361 AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA TTGCTACAGG CATCGTGGTG 3421 TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT CCCAACGATC AAGGCGAGTT 3481 ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT TCGGTCCTCC GATCGTTGTC 3541 AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG CAGCACTGCA TAATTCTCTT 3601 ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG AGTACTCAAC CAAGTCATTC 3661 TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG CGTCAATACG GGATAATACC 3721 GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA AACGTTCTTC GGGGCGAAAA 3781 CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT AACCCACTCG TGCACCCAAC 3841 TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT GAGCAAAAAC AGGAAGGCAA 3901 AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT GAATACTCAT ACTCTTCCTT 3961 TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA TGAGCGGATA CATATTTGAA 4021 TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACCT GCACCAGTCA GTAAAACGAC 4081 GGCCAGTAGT CAAAAGCCTC CGACCGGAGG CTTTTGACTT GGTTCAGGTG GAGTGGGAG //