LOCUS Exported 4721 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..4721 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 38..85 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 1933..1949 /label="M13 for" /note="common sequencing primer, one of multiple similar variants" terminator 1950..1981 /label="tonB terminator" /note="bidirectional E. coli tonB-P14 transcription terminator" CDS 100..4720 /label="topA" /note="Releases the supercoiling and torsional tension of DNA, which is introduced during the DNA replication and transcription, by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(5'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 3'-OH DNA strand. The free DNA strand then undergoes passage around the unbroken strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 3'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone." /translation="MGKALVIVESPAKAKTINKYLGSDYVVKSSVGHIRDLPTSGSAAKKSADSTSTKTAKKPKKDERGALVNRMGVDPWHNWEAHYEVLPGKEKVVSELKQLAEKADHIYLATDLDREGEAIAWHLREVIGGDDARYSRVVFNEITKNAIRQAFNKPGELNIDRVNAQQARRFMDRVVGYMVSPLLWKKIARGLSAGRVQSVAVRLVVEREREIKAFVPEEFWEVDASTTTPSGEALALQVTHQNDKPFRPVNKEQTQAAVSLLEKARYSVLEREDKPTTSKPGAPFITSTLQQAASTRLGFGVKKTMMMAQRLYEAGYITYMRTDSTNLSQDAVNMVRGYISDNFGKKYLPESPNQYASKENSQEAHEAIRPSDVNVMAESLKDMEADAQKLYQLIWRQFVACQMTPAKYDSTTLTVGAGDFRLKARGRILRFDGWTKVMPALRKGDEDRILPAVNKGDALTLVELTPAQHFTKPPARFSEASLVKELEKRGIGRPSTYASIISTIQDRGYVRVENRRFYAEKMGEIVTDRLEENFRELMNYDFTAQMENSLDQVANHEAEWKAVLDHFFSDFTQQLDKAEKDPEEGGMRPNQMVLTSIDCPTCGRKMGIRTASTGVFLGCSGYALPPKERCKTTINLVPENEVLNVLEGEDAETNALRAKRRCPKCGTAMDSYLIDPKRKLHVCGNNPTCDGYEIEEGEFRIKGYDGPIVECEKCGSEMHLKMGRFGKYMACTNEECKNTRKILRNGEVAPPKEDPVPLPELPCEKSDAYFVLRDGAAGVFLAANTFPKSRETRAPLVEELYRFRDRLPEKLRYLADAPQQDPEGNKTMVRFSRKTKQQYVSSEKDGKATGWSAFYVDGKWVEGKK" primer_bind complement(90..106) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(173..761) /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(952..1812) /label="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*" promoter complement(1813..1917) /label="bla promoter" /note="ampicillin resistance gene promoter" misc_feature 4662..4721 /label="FreeGenes Barcode" /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." ORIGIN 1 CGCTGAGGTG TCAATCGTCG GAGCCGCTGA GCAATAACTA GCATAACCCC TTGGGGCCTC 61 TAAACGGGTC TTGAGGGGTT TTTTGCATGG TCATAGCTGT TTCCTGAGAG CTTGGCAGGT 121 GATGACACAC ATTAACAAAT TTCGTGAGGA GTCTCCAGAA GAATGCCATT AATTTCCATA 181 GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC 241 CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG 301 TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC 361 TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG 421 GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC 481 TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA 541 TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG 601 GCTACACTAG AAGAACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA 661 AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG 721 TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGGCCTA CTATTAGCAA 781 CAACGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA 841 AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA 901 ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG 961 CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG 1021 ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC 1081 AATGATACCG CGAGAACCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC 1141 CGGAAGGGCC GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA 1201 TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC 1261 CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG 1321 TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC 1381 CTTCGGTCCT CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT 1441 GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG 1501 TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC 1561 GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG 1621 AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT 1681 GTAACCCACT CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG 1741 GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG 1801 TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT 1861 CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC 1921 CTGCACCAGT CAGTAAAACG ACGGCCAGTA GTCAAAAGCC TCCGACCGGA GGCTTTTGAC 1981 TTGGTTCAGG TGGAGTGGGA GAAAGCACTC TTAGGCCTCT GGGTCTCTAA TGGGAAAAGC 2041 ACTAGTCATC GTCGAATCAC CGGCAAAAGC AAAAACCATC AACAAATACC TAGGATCAGA 2101 CTACGTCGTC AAATCATCAG TCGGACACAT CCGTGACCTA CCGACCTCAG GATCAGCAGC 2161 AAAAAAATCA GCAGACTCAA CCTCAACCAA AACCGCAAAA AAACCGAAAA AAGACGAACG 2221 TGGAGCACTA GTCAACCGTA TGGGAGTCGA CCCGTGGCAC AACTGGGAAG CACACTACGA 2281 AGTCCTACCG GGAAAAGAAA AAGTCGTCTC AGAACTAAAA CAGCTAGCAG AAAAAGCAGA 2341 CCACATCTAC CTAGCAACCG ACCTAGACCG TGAAGGAGAA GCAATCGCAT GGCACCTACG 2401 TGAAGTCATC GGAGGAGACG ACGCACGTTA CTCACGTGTC GTCTTCAACG AAATCACCAA 2461 AAACGCAATC CGTCAGGCAT TCAACAAACC GGGAGAACTA AACATCGACC GTGTCAACGC 2521 ACAGCAGGCA CGTCGTTTCA TGGACCGTGT CGTCGGATAC ATGGTCTCAC CGCTACTATG 2581 GAAAAAAATC GCACGTGGAC TATCAGCAGG ACGTGTCCAG TCAGTCGCAG TCCGTCTAGT 2641 CGTCGAACGT GAACGTGAAA TCAAAGCATT CGTCCCGGAA GAATTCTGGG AAGTCGACGC 2701 ATCAACCACC ACCCCGTCAG GAGAAGCACT AGCACTACAG GTCACCCACC AGAACGACAA 2761 ACCGTTCCGT CCGGTCAACA AAGAACAGAC CCAGGCAGCA GTCTCACTAC TAGAAAAAGC 2821 ACGTTACTCA GTCCTAGAAC GTGAAGACAA ACCGACCACC TCAAAACCGG GAGCACCGTT 2881 CATCACCTCA ACCCTACAGC AGGCAGCATC AACCCGTCTA GGATTCGGAG TCAAAAAAAC 2941 CATGATGATG GCACAGCGTC TATACGAAGC AGGATACATC ACCTACATGC GTACCGACTC 3001 AACCAACCTA TCACAGGACG CAGTCAACAT GGTCCGTGGA TACATCTCAG ACAACTTCGG 3061 AAAAAAATAC CTACCGGAAT CACCGAACCA GTACGCATCA AAAGAAAACT CACAGGAAGC 3121 ACACGAAGCA ATCCGTCCGT CAGACGTCAA CGTCATGGCA GAATCACTAA AAGACATGGA 3181 AGCAGACGCA CAGAAACTAT ACCAGCTAAT CTGGCGTCAG TTCGTCGCAT GCCAGATGAC 3241 CCCGGCAAAA TACGACTCAA CCACCCTAAC CGTCGGAGCA GGAGACTTCC GTCTAAAAGC 3301 ACGTGGACGT ATCCTACGTT TCGACGGATG GACCAAAGTC ATGCCGGCAC TACGTAAAGG 3361 AGACGAAGAC CGTATCCTAC CGGCAGTCAA CAAAGGAGAC GCACTAACCC TAGTCGAACT 3421 AACCCCGGCA CAGCACTTCA CCAAACCGCC GGCACGTTTC TCAGAAGCAT CACTAGTCAA 3481 AGAACTAGAA AAACGTGGAA TCGGACGTCC GTCAACCTAC GCATCAATCA TCTCAACCAT 3541 CCAGGACCGT GGATACGTCC GTGTCGAAAA CCGTCGTTTC TACGCAGAAA AAATGGGAGA 3601 AATCGTCACC GACCGTCTAG AAGAAAACTT CCGTGAACTA ATGAACTACG ACTTCACCGC 3661 ACAGATGGAA AACTCACTAG ACCAGGTCGC AAACCACGAA GCAGAATGGA AAGCAGTCCT 3721 AGACCACTTC TTCTCAGACT TCACCCAGCA GCTAGACAAA GCAGAAAAAG ACCCGGAAGA 3781 AGGAGGAATG CGTCCGAACC AGATGGTCCT AACCTCAATC GACTGCCCGA CCTGCGGACG 3841 TAAAATGGGA ATCCGTACCG CATCAACCGG AGTCTTCCTA GGATGCTCAG GATACGCACT 3901 ACCGCCGAAA GAACGTTGCA AAACCACCAT CAACCTAGTC CCGGAAAACG AAGTCCTAAA 3961 CGTCCTAGAA GGAGAAGACG CAGAAACCAA CGCACTACGT GCAAAACGTC GTTGCCCGAA 4021 ATGCGGAACC GCAATGGACT CATACCTAAT CGACCCGAAA CGTAAACTAC ACGTCTGCGG 4081 AAACAACCCG ACCTGCGACG GATACGAAAT CGAAGAAGGA GAATTCCGTA TCAAAGGATA 4141 CGACGGACCG ATCGTCGAAT GCGAAAAATG CGGATCAGAA ATGCACCTAA AAATGGGACG 4201 TTTCGGAAAA TACATGGCAT GCACCAACGA AGAATGCAAA AACACCCGTA AAATCCTACG 4261 TAACGGAGAA GTCGCACCGC CGAAAGAAGA CCCGGTCCCG CTACCGGAAC TACCGTGCGA 4321 AAAATCAGAC GCATACTTCG TCCTACGTGA CGGAGCAGCA GGAGTCTTCC TAGCAGCAAA 4381 CACCTTCCCG AAATCACGTG AAACCCGTGC ACCGCTAGTC GAAGAACTAT ACCGTTTCCG 4441 TGACCGTCTA CCGGAAAAAC TACGTTACCT AGCAGACGCA CCGCAGCAGG ACCCGGAAGG 4501 AAACAAAACC ATGGTCCGTT TCTCACGTAA AACCAAACAG CAGTACGTCT CATCAGAAAA 4561 AGACGGAAAA GCAACCGGAT GGTCAGCATT CTACGTCGAC GGAAAATGGG TCGAAGGAAA 4621 AAAATGAAGA GCTTCGAGAC CCCAGAGCTT AGGGGACATA CAAGATAAAT CTTGGTATCC 4681 AATAGGGATA ACAGGGTAAT TTGAACACCT CACTGGAATA T //