LOCUS       V015868                 8020 bp    DNA     circular SYN 04-AUG-2020
DEFINITION  Exported.
ACCESSION   V015868
VERSION     V015868
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8020)
  AUTHORS   Inaba Y, West AC, Banta S.
  TITLE     Enhanced microbial corrosion of stainless steel by Acidithiobacillus
            ferrooxidans through the manipulation of substrate oxidation and
            overexpression of rus
  JOURNAL   Biotechnol. Bioeng. (2020) In press
   PUBMED   32687219
REFERENCE   2  (bases 1 to 8020)
  AUTHORS   Inaba Y, West AC, Banta S.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2020) Chemical Engineering, Columbia University,
            500 W 120th St., New York, NY 10025, USA
REFERENCE   3  (bases 1 to 8020)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Bioeng. (2020) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (11-JUN-2020) Chemical Engineering, Columbia University, 500 W 120th
            St., New York, NY 10025, USA"
FEATURES             Location/Qualifiers
     source          1..8020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(329..1120)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        1664..1692
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1700..1716
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1741..2301
                     /gene="rus"
                     /label="Rusticyanin"
                     /note="Rusticyanin from Acidithiobacillus ferrooxidans
                     (strain ATCC 23270 / DSM 14882 / CIP 104768 / NCIMB 8455).
                     Accession#: B7JAQ0"
     CDS             2308..2325
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      2329..2415
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2507..2534
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      complement(2603..2997)
                     /direction=LEFT
                     /label="RSF1010 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     RSF1010; requires the RSF1010 RepA/B/C proteins for
                     replication (Scholz et al., 1989)"
     oriT            3339..3426
                     /label="RSF1010 oriT"
                     /note="origin of transfer of the broad-host-range plasmid
                     RSF1010 (Scholz et al., 1989)"
     CDS             4665..5633
                     /label="RSF1010 RepB"
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             6147..6983
                     /label="RSF1010 RepA"
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             6973..7821
                     /label="RSF1010 RepC"
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
ORIGIN
        1 gctcctgcag ttcgggggca tggatgcgcg gatagccgct gctggtttcc tggatgccga
       61 cggatttgca ctgccggtag aactccgcga ggtcgtccag cctcaggcag cagctgaacc
      121 aactcgcgag gggatcgagc ccggggtggg cgaagaactc cagcatgaga tccccgcgct
      181 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
      241 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcgc
      301 ctcgaggcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
      361 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
      421 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
      481 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
      541 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
      601 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
      661 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
      721 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
      781 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
      841 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
      901 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
      961 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     1021 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     1081 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     1141 tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     1201 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     1261 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     1321 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     1381 tccggggtca gcaccgtttc tgcggactgg ctttctacgt gttccgcttc ctttagcagc
     1441 ccttgcgccc tgagtgcttg cggcagcgtg aagctagcca tatgtcatcg actgcacggt
     1501 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
     1561 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
     1621 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg
     1681 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggagg taaaggatcc
     1741 atgtatacac agaacacgat gaaaaagaac tggtatgtaa ctgttggtgc ggctgcggct
     1801 ctggcggcaa cggtcggcat gggcaccgcg atggccggta ctctggatac cacatggaaa
     1861 gaggcgacgc ttccccaagt aaaggccatg ttggagaaag acaccgggaa agtcagtggt
     1921 gacactgtta cctacagtgg caagactgta catgtggtcg cggcggccgt gctcccggga
     1981 tttccatttc cgagttttga agttcatgac aaaaagaacc cgaccttgga gattcccgcg
     2041 ggggcaacgg tggacgtgac cttcattaac accaacaagg gattcggtca tagttttgac
     2101 atcaccaaaa aaggacctcc ttatgcggtt atgccggtga tcgaccccat tgttgcagga
     2161 acaggattca gtccggtccc aaaagatggc aagttcggat atacggattt tacctggcat
     2221 ccgacggcgg gcacttacta ctacgtatgt cagataccgg ggcatgccgc caccggtatg
     2281 ttcggtaaga tcgttgtcaa gggtacccat catcaccatc accactgaca aataaaacga
     2341 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2401 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2461 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     2521 acggatggcc tttttctaga aagcttgcgg ccgcgatcaa gcaggtgcga cagacgtcat
     2581 actagactag tactaactgt cacgaacccc tgcaataact gtcacgcccc cctgcaataa
     2641 ctgtcacgaa cccctgcaat aactgtcacg cccccaaacc tgcaaaccca gcaggggcgg
     2701 gggctggcgg ggtgttggaa aaatccatcc atgattatct aagaataatc cactaggcgc
     2761 ggttatcagc gcccttgtgg ggcgctgctg cccttgccca atatgcccgg ccagaggccg
     2821 gatagctggt ctattcgctg cgctaggcta cacaccgccc caccgctgcg cggcaggggg
     2881 aaaggcgggc aaagcccgct aaaccccaca ccaaaccccg cagaaatacg ctggagcgct
     2941 tttagccgct ttagcggcct ttccccctac ccgaagggtg ggggcgcgtg tgcagccccg
     3001 cagggcctgt ctcggtcgat cattcagccc ggctcatcct tctggcgtgg cggcagaccg
     3061 aacaaggcgc ggtcgtggtc gcgttcaagg tacgcatcca ttgccgccat gagccgatcc
     3121 tccggccact cgctgctgtt caccttggcc aaaatcatgg cccccaccag caccttgcgc
     3181 cttgtttcgt tcttgcgctc ttgctgctgt tcccttgccc gcacccgctg aatttcggca
     3241 ttgattcgcg ctcgttgttc ttcgagcttg gccagccgat ccgccgcctt gttgctcccc
     3301 ttaaccatct tgacacccca ttgttaatgt gctgtctcgt aggctatcat ggaggcacag
     3361 cggcggcaat cccgacccta ctttgtaggg gagggcgcac ttaccggttt ctcttcgaga
     3421 aactggccta acggccaccc ttcgggcggt gcgctctccg agggccattg catggagccg
     3481 aaaagcaaaa gcaacagcga ggcagcatgg cgatttatca ccttacggcg aaaaccggca
     3541 gcaggtcggg cggccaatcg gccagggcca aggccgacta catccagcgc gaaggcaagt
     3601 atgcccgcga catggatgaa gtcttgcacg ccgaatccgg gcacatgccg gagttcgtcg
     3661 agcggcccgc cgactactgg gatgctgccg acctgtatga acgcgccaat gggcggctgt
     3721 tcaaggaggt cgaatttgcc ctgccggtcg agctgaccct cgaccagcag aaggcgctgg
     3781 cgtccgagtt cgcccagcac ctgaccggtg ccgagcgcct gccgtatacg ctggccatcc
     3841 atgccggtgg cggcgagaac ccgcactgcc acctgatgat ctccgagcgg atcaatgacg
     3901 gcatcgagcg gcccgccgct cagtggttca agcggtacaa cggcaagacc ccggagaagg
     3961 gcggggcaca gaagaccgaa gcgctcaagc ccaaggcatg gcttgagcag acccgcgagg
     4021 catgggccga ccatgccaac cgggcattag agcgggctgg ccacgacgcc cgcattgacc
     4081 acagaacact tgaggcgcag ggcatcgagc gcctgcccgg tgttcacctg gggccgaacg
     4141 tggtggagat ggaaggccgg ggcatccgca ccgaccgggc agacgtggcc ctgaacatcg
     4201 acaccgccaa cgcccagatc atcgacttac aggaataccg ggaggcaata gaccatgaac
     4261 gcaatcgaca gagtgaagaa atccagaggc atcaacgagt tagcggagca gatcgaaccg
     4321 ctggcccaga gcatggcgac actggccgac gaagcccggc aggtcatgag ccagacccag
     4381 caggccagcg aggcgcaggc ggcggagtgg ctgaaagccc agcgccagac aggggcggca
     4441 tgggtggagc tggccaaaga gttgcgggag gtagccgccg aggtgagcag cgccgcgcag
     4501 agcgcccgga gcgcgtcgcg ggggtggcac tggaagctat ggctaaccgt gatgctggct
     4561 tccatgatgc ctacggtggt gctgctgatc gcatcgttgc tcttgctcga cctgacgcca
     4621 ctgacaaccg aggacggctc gatctggctg cgcttggtgg cccgatgaag aacgacagga
     4681 ctttgcaggc cataggccga cagctcaagg ccatgggctg tgagcgcttc gatatcggcg
     4741 tcagggacgc caccaccggc cagatgatga accgggaatg gtcagccgcc gaagtgctcc
     4801 agaacacgcc atggctcaag cggatgaatg cccagggcaa tgacgtgtat atcaggcccg
     4861 ccgagcagga gcggcatggt ctggtgctgg tggacgacct cagcgagttt gacctggatg
     4921 acatgaaagc cgagggccgg gagcctgccc tggtagtgga aaccagcccg aagaactatc
     4981 aggcatgggt caaggtggcc gacgccgcag gcggtgaact tcgggggcag attgcccgga
     5041 cgctggccag cgagtacgac gccgacccgg ccagcgccga cagccgccac tatggccgct
     5101 tggcgggctt caccaaccgc aaggacaagc acaccacccg cgccggttat cagccgtggg
     5161 tgctgctgcg tgaatccaag ggcaagaccg ccaccgctgg cccggcgctg gtgcagcagg
     5221 ctggccagca gatcgagcag gcccagcggc agcaggagaa ggcccgcagg ctggccagcc
     5281 tcgaactgcc cgagcggcag cttagccgcc accggcgcac ggcgctggac gagtaccgca
     5341 gcgagatggc cgggctggtc aagcgcttcg gtgatgacct cagcaagtgc gactttatcg
     5401 ccgcgcagaa gctggccagc cggggccgca gtgccgagga aatcggcaag gccatggccg
     5461 aggccagccc agcgctggca gagcgcaagc ccggccacga agcggattac atcgagcgca
     5521 ccgtcagcaa ggtcatgggt ctgcccagcg tccagcttgc gcgggccgag ctggcacggg
     5581 caccggcacc ccgccagcga ggcatggaca ggggcgggcc agatttcagc atgtagtgct
     5641 tgcgttggta ctcacgcctg ttatactatg agtactcacg cacagaaggg ggttttatgg
     5701 aatacgaaaa aagcgcttca gggtcggtct acctgatcaa aagtgacaag ggctattggt
     5761 tgcccggtgg ctttggttat acgtcaaaca aggccgaggc tggccgcttt tcagtcgctg
     5821 atatggccag ccttaacctt gacggctgca ccttgtcctt gttccgcgaa gacaagcctt
     5881 tcggccccgg caagtttctc ggtgactgat atgaaagacc aaaaggacaa gcagaccggc
     5941 gacctgctgg ccagccctga cgctgtacgc caagcgcgat atgccgagcg catgaaggcc
     6001 aaagggatgc gtcagcgcaa gttctggctg accgacgacg aatacgaggc gctgcgcgag
     6061 tgcctggaag aactcagagc ggcgcagggc gggggtagtg accccgccag cgcctaacca
     6121 ccaactgcct gcaaaggagg caatcaatgg ctacccataa gcctatcaat attctggagg
     6181 cgttcgcagc agcgccgcca ccgctggact acgttttgcc caacatggtg gccggtacgg
     6241 tcggggcgct ggtgtcgccc ggtggtgccg gtaaatccat gctggccctg caactggccg
     6301 cacagattgc aggcgggccg gatctgctgg aggtgggcga actgcccacc ggcccggtga
     6361 tctacctgcc cgccgaagac ccgcccaccg ccattcatca ccgcctgcac gcccttgggg
     6421 cgcacctcag cgccgaggaa cggcaagccg tggctgacgg cctgctgatc cagccgctga
     6481 tcggcagcct gcccaacatc atggccccgg agtggttcga cggcctcaag cgcgccgccg
     6541 agggccgccg cctgatggtg ctggacacgc tgcgccggtt ccacatcgag gaagaaaacg
     6601 ccagcggccc catggcccag gtcatcggtc gcatggaggc catcgccgcc gataccgggt
     6661 gctctatcgt gttcctgcac catgccagca agggcgcggc catgatgggc gcaggcgacc
     6721 agcagcaggc cagccggggc agctcggtac tggtcgataa catccgctgg cagtcctacc
     6781 tgtcgagcat gaccagcgcc gaggccgagg aatggggtgt ggacgacgac cagcgccggt
     6841 tcttcgtccg cttcggtgtg agcaaggcca actatggcgc accgttcgct gatcggtggt
     6901 tcaggcggca tgacggcggg gtgctcaagc ccgccgtgct ggagaggcag cgcaagagca
     6961 agggggtgcc ccgtggtgaa gcctaagaac aagcacagcc tcagccacgt ccggcacgac
     7021 ccggcgcact gtctggcccc cggcctgttc cgtgccctca agcggggcga gcgcaagcgc
     7081 agcaagctgg acgtgacgta tgactacggc gacggcaagc ggatcgagtt cagcggcccg
     7141 gagccgctgg gcgctgatga tctgcgcatc ctgcaagggc tggtggccat ggctgggcct
     7201 aatggcctag tgcttggccc ggaacccaag accgaaggcg gacggcagct ccggctgttc
     7261 ctggaaccca agtgggaggc cgtcaccgct gatgccatgg tggtcaaagg tagctatcgg
     7321 gcgctggcaa aggaaatcgg ggcagaggtc gatagtggtg gggcgctcaa gcacatacag
     7381 gactgcatcg agcgcctttg gaaggtatcc atcatcgccc agaatggccg caagcggcag
     7441 gggtttcggc tgctgtcgga gtacgccagc gacgaggcgg acgggcgcct gtacgtggcc
     7501 ctgaacccct tgatcgcgca ggccgtcatg ggtggcggcc agcatgtgcg catcagcatg
     7561 gacgaggtgc gggcgctgga cagcgaaacc gcccgcctgc tgcaccagcg gctgtgtggc
     7621 tggatcgacc ccggcaaaac cggcaaggct tccatagata ccttgtgcgg ctatgtctgg
     7681 ccgtcagagg ccagtggttc gaccatgcgc aagcgccgcc agcgggtgcg cgaggcgttg
     7741 ccggagctgg tcgcgctggg ctggacggta accgagttcg cggcgggcaa gtacgacatc
     7801 acccggccca aggcggcagg ctgacccccc ccactctatt gtaaacaaga catttttatc
     7861 ttttatattc aatggcttat tttcctgcta attggtaata ccatgaaaaa taccatgctc
     7921 agaaaaggct taacaatatt ttgaaaaatt gcctactgag cgctgccgca cagctccata
     7981 ggccgctttc ctggctttgc ttccagatgt atgctcttct
//
