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
//