LOCUS       62056_23125        7475 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Cloning vector pYI39, complete sequence.
ACCESSION   MZ420731
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7475)
  AUTHORS   Inaba Y, West AC, Banta S.
  TITLE     Glutathione synthetase overexpression in Acidithiobacillus 
            ferrooxidans improves halotolerance of iron oxidation
  JOURNAL   Appl Environ Microbiol, AEM0151821 (2021) In press
  PUBMED    34347521
REFERENCE   2  (bases 1 to 7475)
  AUTHORS   Inaba Y, West AC, Banta S.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2021) Chemical Engineering, Columbia University, 
            500 W 120th St., New York, NY 10027, USA
REFERENCE   3  (bases 1 to 7475)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol, AEM0151821 (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JUN-2021) Chemical Engineering, Columbia University, 500 W 120th
            St., New York, NY 10027, USA"
FEATURES             Location/Qualifiers
     source          1..7475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(351..1142)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        1686..1714
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1722..1738
                     /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             1785..1802
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1806..1892
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1984..2011
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      complement(2080..2474)
                     /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            2816..2903
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4142..5110
                     /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             5624..6460
                     /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             6450..7298
                     /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 gcttccagat gtatgctctt ctgctcctgc agttcggggg catggatgcg cggatagccg
       61 ctgctggttt cctggatgcc gacggatttg cactgccggt agaactccgc gaggtcgtcc
      121 agcctcaggc agcagctgaa ccaactcgcg aggggatcga gcccggggtg ggcgaagaac
      181 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
      241 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
      301 gtcgcttggt cggtcatttc gcctcgaggc gaaccccaga gtcccgctca gaagaactcg
      361 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
      421 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
      481 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
      541 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
      601 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
      661 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
      721 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
      781 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
      841 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
      901 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
      961 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     1021 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     1081 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     1141 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
     1201 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     1261 gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc
     1321 catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag
     1381 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     1441 gtgttccgct tcctttagca gcccttgcgc cctgagtgct tgcggcagcg tgaagctagc
     1501 catatgtcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct
     1561 gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc
     1621 cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg
     1681 agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt
     1741 tcacacagga ggtaaaggat ccccttagaa ttcgttctgg tacccatcat caccatcacc
     1801 actgacaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1861 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     1921 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     1981 agcagaaggc catcctgacg gatggccttt ttctagaaag cttgcggccg cgatcaagca
     2041 ggtgcgacag acgtcatact agactagtac taactgtcac gaacccctgc aataactgtc
     2101 acgcccccct gcaataactg tcacgaaccc ctgcaataac tgtcacgccc ccaaacctgc
     2161 aaacccagca ggggcggggg ctggcggggt gttggaaaaa tccatccatg attatctaag
     2221 aataatccac taggcgcggt tatcagcgcc cttgtggggc gctgctgccc ttgcccaata
     2281 tgcccggcca gaggccggat agctggtcta ttcgctgcgc taggctacac accgccccac
     2341 cgctgcgcgg cagggggaaa ggcgggcaaa gcccgctaaa ccccacacca aaccccgcag
     2401 aaatacgctg gagcgctttt agccgcttta gcggcctttc cccctacccg aagggtgggg
     2461 gcgcgtgtgc agccccgcag ggcctgtctc ggtcgatcat tcagcccggc tcatccttct
     2521 ggcgtggcgg cagaccgaac aaggcgcggt cgtggtcgcg ttcaaggtac gcatccattg
     2581 ccgccatgag ccgatcctcc ggccactcgc tgctgttcac cttggccaaa atcatggccc
     2641 ccaccagcac cttgcgcctt gtttcgttct tgcgctcttg ctgctgttcc cttgcccgca
     2701 cccgctgaat ttcggcattg attcgcgctc gttgttcttc gagcttggcc agccgatccg
     2761 ccgccttgtt gctcccctta accatcttga caccccattg ttaatgtgct gtctcgtagg
     2821 ctatcatgga ggcacagcgg cggcaatccc gaccctactt tgtaggggag ggcgcactta
     2881 ccggtttctc ttcgagaaac tggcctaacg gccacccttc gggcggtgcg ctctccgagg
     2941 gccattgcat ggagccgaaa agcaaaagca acagcgaggc agcatggcga tttatcacct
     3001 tacggcgaaa accggcagca ggtcgggcgg ccaatcggcc agggccaagg ccgactacat
     3061 ccagcgcgaa ggcaagtatg cccgcgacat ggatgaagtc ttgcacgccg aatccgggca
     3121 catgccggag ttcgtcgagc ggcccgccga ctactgggat gctgccgacc tgtatgaacg
     3181 cgccaatggg cggctgttca aggaggtcga atttgccctg ccggtcgagc tgaccctcga
     3241 ccagcagaag gcgctggcgt ccgagttcgc ccagcacctg accggtgccg agcgcctgcc
     3301 gtatacgctg gccatccatg ccggtggcgg cgagaacccg cactgccacc tgatgatctc
     3361 cgagcggatc aatgacggca tcgagcggcc cgccgctcag tggttcaagc ggtacaacgg
     3421 caagaccccg gagaagggcg gggcacagaa gaccgaagcg ctcaagccca aggcatggct
     3481 tgagcagacc cgcgaggcat gggccgacca tgccaaccgg gcattagagc gggctggcca
     3541 cgacgcccgc attgaccaca gaacacttga ggcgcagggc atcgagcgcc tgcccggtgt
     3601 tcacctgggg ccgaacgtgg tggagatgga aggccggggc atccgcaccg accgggcaga
     3661 cgtggccctg aacatcgaca ccgccaacgc ccagatcatc gacttacagg aataccggga
     3721 ggcaatagac catgaacgca atcgacagag tgaagaaatc cagaggcatc aacgagttag
     3781 cggagcagat cgaaccgctg gcccagagca tggcgacact ggccgacgaa gcccggcagg
     3841 tcatgagcca gacccagcag gccagcgagg cgcaggcggc ggagtggctg aaagcccagc
     3901 gccagacagg ggcggcatgg gtggagctgg ccaaagagtt gcgggaggta gccgccgagg
     3961 tgagcagcgc cgcgcagagc gcccggagcg cgtcgcgggg gtggcactgg aagctatggc
     4021 taaccgtgat gctggcttcc atgatgccta cggtggtgct gctgatcgca tcgttgctct
     4081 tgctcgacct gacgccactg acaaccgagg acggctcgat ctggctgcgc ttggtggccc
     4141 gatgaagaac gacaggactt tgcaggccat aggccgacag ctcaaggcca tgggctgtga
     4201 gcgcttcgat atcggcgtca gggacgccac caccggccag atgatgaacc gggaatggtc
     4261 agccgccgaa gtgctccaga acacgccatg gctcaagcgg atgaatgccc agggcaatga
     4321 cgtgtatatc aggcccgccg agcaggagcg gcatggtctg gtgctggtgg acgacctcag
     4381 cgagtttgac ctggatgaca tgaaagccga gggccgggag cctgccctgg tagtggaaac
     4441 cagcccgaag aactatcagg catgggtcaa ggtggccgac gccgcaggcg gtgaacttcg
     4501 ggggcagatt gcccggacgc tggccagcga gtacgacgcc gacccggcca gcgccgacag
     4561 ccgccactat ggccgcttgg cgggcttcac caaccgcaag gacaagcaca ccacccgcgc
     4621 cggttatcag ccgtgggtgc tgctgcgtga atccaagggc aagaccgcca ccgctggccc
     4681 ggcgctggtg cagcaggctg gccagcagat cgagcaggcc cagcggcagc aggagaaggc
     4741 ccgcaggctg gccagcctcg aactgcccga gcggcagctt agccgccacc ggcgcacggc
     4801 gctggacgag taccgcagcg agatggccgg gctggtcaag cgcttcggtg atgacctcag
     4861 caagtgcgac tttatcgccg cgcagaagct ggccagccgg ggccgcagtg ccgaggaaat
     4921 cggcaaggcc atggccgagg ccagcccagc gctggcagag cgcaagcccg gccacgaagc
     4981 ggattacatc gagcgcaccg tcagcaaggt catgggtctg cccagcgtcc agcttgcgcg
     5041 ggccgagctg gcacgggcac cggcaccccg ccagcgaggc atggacaggg gcgggccaga
     5101 tttcagcatg tagtgcttgc gttggtactc acgcctgtta tactatgagt actcacgcac
     5161 agaagggggt tttatggaat acgaaaaaag cgcttcaggg tcggtctacc tgatcaaaag
     5221 tgacaagggc tattggttgc ccggtggctt tggttatacg tcaaacaagg ccgaggctgg
     5281 ccgcttttca gtcgctgata tggccagcct taaccttgac ggctgcacct tgtccttgtt
     5341 ccgcgaagac aagcctttcg gccccggcaa gtttctcggt gactgatatg aaagaccaaa
     5401 aggacaagca gaccggcgac ctgctggcca gccctgacgc tgtacgccaa gcgcgatatg
     5461 ccgagcgcat gaaggccaaa gggatgcgtc agcgcaagtt ctggctgacc gacgacgaat
     5521 acgaggcgct gcgcgagtgc ctggaagaac tcagagcggc gcagggcggg ggtagtgacc
     5581 ccgccagcgc ctaaccacca actgcctgca aaggaggcaa tcaatggcta cccataagcc
     5641 tatcaatatt ctggaggcgt tcgcagcagc gccgccaccg ctggactacg ttttgcccaa
     5701 catggtggcc ggtacggtcg gggcgctggt gtcgcccggt ggtgccggta aatccatgct
     5761 ggccctgcaa ctggccgcac agattgcagg cgggccggat ctgctggagg tgggcgaact
     5821 gcccaccggc ccggtgatct acctgcccgc cgaagacccg cccaccgcca ttcatcaccg
     5881 cctgcacgcc cttggggcgc acctcagcgc cgaggaacgg caagccgtgg ctgacggcct
     5941 gctgatccag ccgctgatcg gcagcctgcc caacatcatg gccccggagt ggttcgacgg
     6001 cctcaagcgc gccgccgagg gccgccgcct gatggtgctg gacacgctgc gccggttcca
     6061 catcgaggaa gaaaacgcca gcggccccat ggcccaggtc atcggtcgca tggaggccat
     6121 cgccgccgat accgggtgct ctatcgtgtt cctgcaccat gccagcaagg gcgcggccat
     6181 gatgggcgca ggcgaccagc agcaggccag ccggggcagc tcggtactgg tcgataacat
     6241 ccgctggcag tcctacctgt cgagcatgac cagcgccgag gccgaggaat ggggtgtgga
     6301 cgacgaccag cgccggttct tcgtccgctt cggtgtgagc aaggccaact atggcgcacc
     6361 gttcgctgat cggtggttca ggcggcatga cggcggggtg ctcaagcccg ccgtgctgga
     6421 gaggcagcgc aagagcaagg gggtgccccg tggtgaagcc taagaacaag cacagcctca
     6481 gccacgtccg gcacgacccg gcgcactgtc tggcccccgg cctgttccgt gccctcaagc
     6541 ggggcgagcg caagcgcagc aagctggacg tgacgtatga ctacggcgac ggcaagcgga
     6601 tcgagttcag cggcccggag ccgctgggcg ctgatgatct gcgcatcctg caagggctgg
     6661 tggccatggc tgggcctaat ggcctagtgc ttggcccgga acccaagacc gaaggcggac
     6721 ggcagctccg gctgttcctg gaacccaagt gggaggccgt caccgctgat gccatggtgg
     6781 tcaaaggtag ctatcgggcg ctggcaaagg aaatcggggc agaggtcgat agtggtgggg
     6841 cgctcaagca catacaggac tgcatcgagc gcctttggaa ggtatccatc atcgcccaga
     6901 atggccgcaa gcggcagggg tttcggctgc tgtcggagta cgccagcgac gaggcggacg
     6961 ggcgcctgta cgtggccctg aaccccttga tcgcgcaggc cgtcatgggt ggcggccagc
     7021 atgtgcgcat cagcatggac gaggtgcggg cgctggacag cgaaaccgcc cgcctgctgc
     7081 accagcggct gtgtggctgg atcgaccccg gcaaaaccgg caaggcttcc atagatacct
     7141 tgtgcggcta tgtctggccg tcagaggcca gtggttcgac catgcgcaag cgccgccagc
     7201 gggtgcgcga ggcgttgccg gagctggtcg cgctgggctg gacggtaacc gagttcgcgg
     7261 cgggcaagta cgacatcacc cggcccaagg cggcaggctg acccccccca ctctattgta
     7321 aacaagacat ttttatcttt tatattcaat ggcttatttt cctgctaatt ggtaatacca
     7381 tgaaaaatac catgctcaga aaaggcttaa caatattttg aaaaattgcc tactgagcgc
     7441 tgccgcacag ctccataggc cgctttcctg gcttt
//
