LOCUS       Exported     4538 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..4538
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
 polymerase"
     terminator      38..85
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
                     /label="T7 terminator"
     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             2030..4444
                     /label="gyrB"
                     /note="DNA gyrase negatively supercoils closed circular double-stranded DNA in an ATP-dependent manner to maintain chromosomes in an underwound state (PubMed:186775, PubMed:3031051, PubMed:1323022, PubMed:8248233, PubMed:7811004, PubMed:8621650, PubMed:9657678, PubMed:12051842, PubMed:12051843, PubMed:18642932, PubMed:19060136, PubMed:19965760, PubMed:22457353, PubMed:23294697, PubMed:20356737, PubMed:20675723, PubMed:23352267, PubMed:24386374, PubMed:25202966, PubMed:25849408). This makes better substrates for topoisomerase 4 (ParC and ParE) which is the main enzyme that unlinks newly replicated chromosomes in E.coli (PubMed:9334322). Gyrase catalyzes the interconversion of other topological isomers of double-stranded DNA rings, including catenanes (PubMed:22457352). Relaxes negatively supercoiled DNA in an ATP-independent manner (PubMed:337300). E.coli gyrase has higher supercoiling activity than other characterized bacterial gyrases; at comparable concentrations E.coli gyrase introduces more supercoils faster than M.tuberculosis gyrase, while M.tuberculosis gyrase has higher decatenation than supercoiling activity compared to E.coli (PubMed:22457352). E.coli makes 15% more negative supercoils in pBR322 plasmid DNA than S.typhimurium; the S.typhimurium GyrB subunit is toxic in E.coli, while the E.coli copy can be expressed in S.typhimurium even though the 2 subunits have 777/804 residues identical (PubMed:17400739). The enzymatic differences between E.coli gyrase and topoisomerase IV are largely due to the GyrA C-terminal domain (approximately residues 524-841) and specifically the GyrA-box (PubMed:8962066, PubMed:16332690)."
                     /translation="MSNSYDSSSIKVLKGLDAVRKRPGMYIGDTDDGTGLHHMVFEVVDNAIDEALAGHCKEIIVTIHADNSVSVQDDGRGIPTGIHPEEGVSAAEVIMTVLHAGGKFDDNSYKVSGGLHGVGVSVVNALSQKLELVIQREGKIHRQIYEHGVPQAPLAVTGETEKTGTMVRFWPSLETFTNVTEFEYEILAKRLRELSFLNSGVSIRLRDKRDGKEDHFHYEGGIKAFVEYLNKNKTPIHPNIFYFSTEKDGIGVEVALQWNDGFQENIYCFTNNIPQRDGGTHLAGFRAAMTRTLNAYMDKEGYSKKAKVSATGDDAREGLIAVVSVKVPDPKFSSQTKDKLVSSEVKSAVEQQMNELLAEYLLENPTDAKIVVGKIIDAARAREAARRAREMTRRKGALDLAGLPGKLADCQERDPALSELYLVEGDSAGGSAKQGRNRKNQAILPLKGKILNVEKARFDKMLSSQEVATLITALGCGIGRDEYNPDKLRYHSIIIMTDADVDGSHIRTLLLTFFYRQMPEIVERGHVYIAQPPLYKVKKGKQEQYIKDDEAMDQYQISIALDGATLHTNASAPALAGEALEKLVSEYNATQKMINRMERRYPKAMLKELIYQPTLTEADLSDEQTVTRWVNALVSELNDKEQHGSQWKFDVHTNAEQNLFEPIVRVRTHGVDTDYPLDHEFITGGEYRRICTLGEKLRGLLEEDAFIERGERRQPVASFEQALDWLVKESRRGLSIQRYKGLGEMNPEQLWETTMDPESRRMLRVTVKDAIAADQLFTTLMGDAVEPRRAFIEENALKAANIDI"
     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    4479..4538
                     /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 GAAACACGTG GCAAACATTC CGGTCTCTAA TGTCAAACTC
     2041 ATACGACTCA TCATCAATCA AAGTCCTAAA AGGACTAGAC GCAGTCCGTA AACGTCCGGG
     2101 AATGTACATC GGAGACACCG ACGACGGAAC CGGACTACAC CACATGGTCT TCGAAGTCGT
     2161 CGACAACGCA ATCGACGAAG CACTAGCAGG ACACTGCAAA GAAATCATCG TCACCATCCA
     2221 CGCAGACAAC TCAGTCTCAG TCCAGGACGA CGGACGTGGA ATCCCGACCG GAATCCACCC
     2281 GGAAGAAGGA GTCTCAGCAG CAGAAGTCAT CATGACCGTC CTACACGCAG GAGGAAAATT
     2341 CGACGACAAC TCATACAAAG TCTCAGGAGG ACTACACGGA GTCGGAGTCT CAGTCGTCAA
     2401 CGCACTATCA CAGAAACTAG AACTAGTCAT CCAGCGTGAA GGAAAAATCC ACCGTCAGAT
     2461 CTACGAACAC GGAGTCCCGC AGGCACCGCT AGCAGTCACC GGAGAAACCG AAAAAACCGG
     2521 AACCATGGTC CGTTTCTGGC CGTCACTAGA AACCTTCACC AACGTCACCG AATTCGAATA
     2581 CGAAATCCTA GCAAAACGTC TACGTGAACT ATCATTCCTA AACTCAGGAG TCTCAATCCG
     2641 TCTACGTGAC AAACGTGACG GAAAAGAAGA CCACTTCCAC TACGAAGGAG GAATCAAAGC
     2701 ATTCGTCGAA TACCTAAACA AAAACAAAAC CCCGATCCAC CCGAACATCT TCTACTTCTC
     2761 AACCGAAAAA GACGGAATCG GAGTCGAAGT CGCACTACAG TGGAACGACG GATTCCAGGA
     2821 AAACATCTAC TGCTTCACCA ACAACATCCC GCAGCGTGAC GGAGGAACCC ACCTAGCAGG
     2881 ATTCCGTGCA GCAATGACCC GTACCCTAAA CGCATACATG GACAAAGAAG GATACTCAAA
     2941 AAAAGCAAAA GTCTCAGCAA CCGGAGACGA CGCACGTGAA GGACTAATCG CAGTCGTCTC
     3001 AGTCAAAGTC CCGGACCCGA AATTCTCATC ACAGACCAAA GACAAACTAG TCTCATCAGA
     3061 AGTCAAATCA GCAGTCGAAC AGCAGATGAA CGAACTACTA GCAGAATACC TACTAGAAAA
     3121 CCCGACCGAC GCAAAAATCG TCGTCGGAAA AATCATCGAC GCAGCACGTG CACGTGAAGC
     3181 AGCACGTCGT GCACGTGAAA TGACCCGTCG TAAAGGAGCA CTAGACCTAG CAGGACTACC
     3241 GGGAAAACTA GCAGACTGCC AGGAACGTGA CCCGGCACTA TCAGAACTAT ACCTAGTCGA
     3301 AGGAGACTCA GCAGGAGGAT CAGCAAAACA GGGACGTAAC CGTAAAAACC AGGCAATCCT
     3361 ACCGCTAAAA GGAAAAATCC TAAACGTCGA AAAAGCACGT TTCGACAAAA TGCTATCATC
     3421 ACAGGAAGTC GCAACCCTAA TCACCGCACT AGGATGCGGA ATCGGACGTG ACGAATACAA
     3481 CCCGGACAAA CTACGTTACC ACTCAATCAT CATCATGACC GACGCAGACG TCGACGGATC
     3541 ACACATCCGT ACCCTACTAC TAACCTTCTT CTACCGTCAG ATGCCGGAAA TCGTCGAACG
     3601 TGGACACGTC TACATCGCAC AGCCGCCGCT ATACAAAGTC AAAAAAGGAA AACAGGAACA
     3661 GTACATCAAA GACGACGAAG CAATGGACCA GTACCAGATC TCAATCGCAC TAGACGGAGC
     3721 AACCCTACAC ACCAACGCAT CAGCACCGGC ACTAGCAGGA GAAGCACTAG AAAAACTAGT
     3781 CTCAGAATAC AACGCAACCC AGAAAATGAT CAACCGTATG GAACGTCGTT ACCCGAAAGC
     3841 AATGCTAAAA GAACTAATCT ACCAGCCGAC CCTAACCGAA GCAGACCTAT CAGACGAACA
     3901 GACCGTCACC CGTTGGGTCA ACGCACTAGT CTCAGAACTA AACGACAAAG AACAGCACGG
     3961 ATCACAGTGG AAATTCGACG TCCACACCAA CGCAGAACAG AACCTATTCG AACCGATCGT
     4021 CCGTGTCCGT ACCCACGGAG TCGACACCGA CTACCCGCTA GACCACGAAT TCATCACCGG
     4081 AGGAGAATAC CGTCGTATCT GCACCCTAGG AGAAAAACTA CGTGGACTAC TAGAAGAAGA
     4141 CGCATTCATC GAACGTGGAG AACGTCGTCA GCCGGTCGCA TCATTCGAAC AGGCACTAGA
     4201 CTGGCTAGTC AAAGAATCAC GTCGTGGACT ATCAATCCAG CGTTACAAAG GACTAGGAGA
     4261 AATGAACCCG GAACAGCTAT GGGAAACCAC CATGGACCCG GAATCACGTC GTATGCTACG
     4321 TGTCACCGTC AAAGACGCAA TCGCAGCAGA CCAGCTATTC ACCACCCTAA TGGGAGACGC
     4381 AGTCGAACCG CGTCGTGCAT TCATCGAAGA AAACGCACTA AAAGCAGCAA ACATCGACAT
     4441 CTGAAGAGCT TCGAGACCAT TCAAGGGTTG GACGACTCAG CTCAGCCAAT TATTTCGCTT
     4501 AGGGATAACA GGGTAATCGT ACTAGCACTA CACTTTAG
//