LOCUS       62056_23115        7454 bp DNA     circular SYN 04-AUG-2020
DEFINITION  Cloning vector pYI11, complete sequence.
ACCESSION   MT603571
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  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 7454)
  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 7454)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Bioeng. (2020) In press"
COMMENT     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..7454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(329..1120)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     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)."
     terminator      1763..1849
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1941..1968
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      complement(2037..2431)
                     /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            2773..2860
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4099..5067
                     /codon_start=1
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             5581..6417
                     /codon_start=1
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             6407..7255
                     /codon_start=1
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
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 ccttagaatt cgttctggta cccaaataaa acgaaaggct cagtcgaaag actgggcctt
     1801 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     1861 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     1921 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttc tagaaagctt
     1981 gcggccgcga tcaagcaggt gcgacagacg tcatactaga ctagtactaa ctgtcacgaa
     2041 cccctgcaat aactgtcacg cccccctgca ataactgtca cgaacccctg caataactgt
     2101 cacgccccca aacctgcaaa cccagcaggg gcgggggctg gcggggtgtt ggaaaaatcc
     2161 atccatgatt atctaagaat aatccactag gcgcggttat cagcgccctt gtggggcgct
     2221 gctgcccttg cccaatatgc ccggccagag gccggatagc tggtctattc gctgcgctag
     2281 gctacacacc gccccaccgc tgcgcggcag ggggaaaggc gggcaaagcc cgctaaaccc
     2341 cacaccaaac cccgcagaaa tacgctggag cgcttttagc cgctttagcg gcctttcccc
     2401 ctacccgaag ggtgggggcg cgtgtgcagc cccgcagggc ctgtctcggt cgatcattca
     2461 gcccggctca tccttctggc gtggcggcag accgaacaag gcgcggtcgt ggtcgcgttc
     2521 aaggtacgca tccattgccg ccatgagccg atcctccggc cactcgctgc tgttcacctt
     2581 ggccaaaatc atggccccca ccagcacctt gcgccttgtt tcgttcttgc gctcttgctg
     2641 ctgttccctt gcccgcaccc gctgaatttc ggcattgatt cgcgctcgtt gttcttcgag
     2701 cttggccagc cgatccgccg ccttgttgct ccccttaacc atcttgacac cccattgtta
     2761 atgtgctgtc tcgtaggcta tcatggaggc acagcggcgg caatcccgac cctactttgt
     2821 aggggagggc gcacttaccg gtttctcttc gagaaactgg cctaacggcc acccttcggg
     2881 cggtgcgctc tccgagggcc attgcatgga gccgaaaagc aaaagcaaca gcgaggcagc
     2941 atggcgattt atcaccttac ggcgaaaacc ggcagcaggt cgggcggcca atcggccagg
     3001 gccaaggccg actacatcca gcgcgaaggc aagtatgccc gcgacatgga tgaagtcttg
     3061 cacgccgaat ccgggcacat gccggagttc gtcgagcggc ccgccgacta ctgggatgct
     3121 gccgacctgt atgaacgcgc caatgggcgg ctgttcaagg aggtcgaatt tgccctgccg
     3181 gtcgagctga ccctcgacca gcagaaggcg ctggcgtccg agttcgccca gcacctgacc
     3241 ggtgccgagc gcctgccgta tacgctggcc atccatgccg gtggcggcga gaacccgcac
     3301 tgccacctga tgatctccga gcggatcaat gacggcatcg agcggcccgc cgctcagtgg
     3361 ttcaagcggt acaacggcaa gaccccggag aagggcgggg cacagaagac cgaagcgctc
     3421 aagcccaagg catggcttga gcagacccgc gaggcatggg ccgaccatgc caaccgggca
     3481 ttagagcggg ctggccacga cgcccgcatt gaccacagaa cacttgaggc gcagggcatc
     3541 gagcgcctgc ccggtgttca cctggggccg aacgtggtgg agatggaagg ccggggcatc
     3601 cgcaccgacc gggcagacgt ggccctgaac atcgacaccg ccaacgccca gatcatcgac
     3661 ttacaggaat accgggaggc aatagaccat gaacgcaatc gacagagtga agaaatccag
     3721 aggcatcaac gagttagcgg agcagatcga accgctggcc cagagcatgg cgacactggc
     3781 cgacgaagcc cggcaggtca tgagccagac ccagcaggcc agcgaggcgc aggcggcgga
     3841 gtggctgaaa gcccagcgcc agacaggggc ggcatgggtg gagctggcca aagagttgcg
     3901 ggaggtagcc gccgaggtga gcagcgccgc gcagagcgcc cggagcgcgt cgcgggggtg
     3961 gcactggaag ctatggctaa ccgtgatgct ggcttccatg atgcctacgg tggtgctgct
     4021 gatcgcatcg ttgctcttgc tcgacctgac gccactgaca accgaggacg gctcgatctg
     4081 gctgcgcttg gtggcccgat gaagaacgac aggactttgc aggccatagg ccgacagctc
     4141 aaggccatgg gctgtgagcg cttcgatatc ggcgtcaggg acgccaccac cggccagatg
     4201 atgaaccggg aatggtcagc cgccgaagtg ctccagaaca cgccatggct caagcggatg
     4261 aatgcccagg gcaatgacgt gtatatcagg cccgccgagc aggagcggca tggtctggtg
     4321 ctggtggacg acctcagcga gtttgacctg gatgacatga aagccgaggg ccgggagcct
     4381 gccctggtag tggaaaccag cccgaagaac tatcaggcat gggtcaaggt ggccgacgcc
     4441 gcaggcggtg aacttcgggg gcagattgcc cggacgctgg ccagcgagta cgacgccgac
     4501 ccggccagcg ccgacagccg ccactatggc cgcttggcgg gcttcaccaa ccgcaaggac
     4561 aagcacacca cccgcgccgg ttatcagccg tgggtgctgc tgcgtgaatc caagggcaag
     4621 accgccaccg ctggcccggc gctggtgcag caggctggcc agcagatcga gcaggcccag
     4681 cggcagcagg agaaggcccg caggctggcc agcctcgaac tgcccgagcg gcagcttagc
     4741 cgccaccggc gcacggcgct ggacgagtac cgcagcgaga tggccgggct ggtcaagcgc
     4801 ttcggtgatg acctcagcaa gtgcgacttt atcgccgcgc agaagctggc cagccggggc
     4861 cgcagtgccg aggaaatcgg caaggccatg gccgaggcca gcccagcgct ggcagagcgc
     4921 aagcccggcc acgaagcgga ttacatcgag cgcaccgtca gcaaggtcat gggtctgccc
     4981 agcgtccagc ttgcgcgggc cgagctggca cgggcaccgg caccccgcca gcgaggcatg
     5041 gacaggggcg ggccagattt cagcatgtag tgcttgcgtt ggtactcacg cctgttatac
     5101 tatgagtact cacgcacaga agggggtttt atggaatacg aaaaaagcgc ttcagggtcg
     5161 gtctacctga tcaaaagtga caagggctat tggttgcccg gtggctttgg ttatacgtca
     5221 aacaaggccg aggctggccg cttttcagtc gctgatatgg ccagccttaa ccttgacggc
     5281 tgcaccttgt ccttgttccg cgaagacaag cctttcggcc ccggcaagtt tctcggtgac
     5341 tgatatgaaa gaccaaaagg acaagcagac cggcgacctg ctggccagcc ctgacgctgt
     5401 acgccaagcg cgatatgccg agcgcatgaa ggccaaaggg atgcgtcagc gcaagttctg
     5461 gctgaccgac gacgaatacg aggcgctgcg cgagtgcctg gaagaactca gagcggcgca
     5521 gggcgggggt agtgaccccg ccagcgccta accaccaact gcctgcaaag gaggcaatca
     5581 atggctaccc ataagcctat caatattctg gaggcgttcg cagcagcgcc gccaccgctg
     5641 gactacgttt tgcccaacat ggtggccggt acggtcgggg cgctggtgtc gcccggtggt
     5701 gccggtaaat ccatgctggc cctgcaactg gccgcacaga ttgcaggcgg gccggatctg
     5761 ctggaggtgg gcgaactgcc caccggcccg gtgatctacc tgcccgccga agacccgccc
     5821 accgccattc atcaccgcct gcacgccctt ggggcgcacc tcagcgccga ggaacggcaa
     5881 gccgtggctg acggcctgct gatccagccg ctgatcggca gcctgcccaa catcatggcc
     5941 ccggagtggt tcgacggcct caagcgcgcc gccgagggcc gccgcctgat ggtgctggac
     6001 acgctgcgcc ggttccacat cgaggaagaa aacgccagcg gccccatggc ccaggtcatc
     6061 ggtcgcatgg aggccatcgc cgccgatacc gggtgctcta tcgtgttcct gcaccatgcc
     6121 agcaagggcg cggccatgat gggcgcaggc gaccagcagc aggccagccg gggcagctcg
     6181 gtactggtcg ataacatccg ctggcagtcc tacctgtcga gcatgaccag cgccgaggcc
     6241 gaggaatggg gtgtggacga cgaccagcgc cggttcttcg tccgcttcgg tgtgagcaag
     6301 gccaactatg gcgcaccgtt cgctgatcgg tggttcaggc ggcatgacgg cggggtgctc
     6361 aagcccgccg tgctggagag gcagcgcaag agcaaggggg tgccccgtgg tgaagcctaa
     6421 gaacaagcac agcctcagcc acgtccggca cgacccggcg cactgtctgg cccccggcct
     6481 gttccgtgcc ctcaagcggg gcgagcgcaa gcgcagcaag ctggacgtga cgtatgacta
     6541 cggcgacggc aagcggatcg agttcagcgg cccggagccg ctgggcgctg atgatctgcg
     6601 catcctgcaa gggctggtgg ccatggctgg gcctaatggc ctagtgcttg gcccggaacc
     6661 caagaccgaa ggcggacggc agctccggct gttcctggaa cccaagtggg aggccgtcac
     6721 cgctgatgcc atggtggtca aaggtagcta tcgggcgctg gcaaaggaaa tcggggcaga
     6781 ggtcgatagt ggtggggcgc tcaagcacat acaggactgc atcgagcgcc tttggaaggt
     6841 atccatcatc gcccagaatg gccgcaagcg gcaggggttt cggctgctgt cggagtacgc
     6901 cagcgacgag gcggacgggc gcctgtacgt ggccctgaac cccttgatcg cgcaggccgt
     6961 catgggtggc ggccagcatg tgcgcatcag catggacgag gtgcgggcgc tggacagcga
     7021 aaccgcccgc ctgctgcacc agcggctgtg tggctggatc gaccccggca aaaccggcaa
     7081 ggcttccata gataccttgt gcggctatgt ctggccgtca gaggccagtg gttcgaccat
     7141 gcgcaagcgc cgccagcggg tgcgcgaggc gttgccggag ctggtcgcgc tgggctggac
     7201 ggtaaccgag ttcgcggcgg gcaagtacga catcacccgg cccaaggcgg caggctgacc
     7261 ccccccactc tattgtaaac aagacatttt tatcttttat attcaatggc ttattttcct
     7321 gctaattggt aataccatga aaaataccat gctcagaaaa ggcttaacaa tattttgaaa
     7381 aattgcctac tgagcgctgc cgcacagctc cataggccgc tttcctggct ttgcttccag
     7441 atgtatgctc ttct
//
