LOCUS       62056_23130        8767 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Cloning vector pYI43, complete sequence.
ACCESSION   MZ420732
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8767)
  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 8767)
  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 8767)
  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..8767
                     /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             3077..3094
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3098..3184
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3276..3303
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      complement(3372..3766)
                     /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            4108..4195
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             5434..6402
                     /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             6916..7752
                     /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             7742..8590
                     /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 ccatgacgga agtgatacgg gatattcctt tcctggccac
     1801 ggatcgggtg gaggctctgc tggatatcga gcgccacctg atcgtcgagc aatccacgat
     1861 tgagcgctgg tttcggggac aatggcaacg gacgccaccg cctttctatg cctctgtgga
     1921 cttgcgcaat gccggtttca agttggcgcc cgtggacacc aatctgtttt cggcgggctt
     1981 taataacctc aatccggagt tttcggcgct ctatgtttcg gctattcagc attatctgaa
     2041 ccagtaccat ccaggtctgg agcgggtact gctgattcct gaaaaccaca cgcgtaacct
     2101 tttttatctg gagagcgtgg cacgattgcg tgagttgttg gaactcgccg gattggacgt
     2161 gcgagtgggc tccctgattc tggaagaccg tacggtctat gatctgcctt caggtggcca
     2221 gttgctgctg gaaccactgt gtcgcgacga cgggcggctg tcgacgaggg gttttgatgc
     2281 acaactgatc ttactgaaca atgatttatc tggtggtgtg ccggaaatat tggcaggcct
     2341 ggagcaacct attctgcccc ccctcgcggc aggttggcgt aaccggcgta aatcccagca
     2401 cttcacccat taccgtgcgg tggcacagga gctggcggag gtgattggca tcgatccctg
     2461 gctgattgac cccgtttttc ggcgttgtca gggcattgac ttcatgcgct ccgaaggccg
     2521 cgagtgcctg gttgcgaacg tggatgccgt actggagatc acccgcgagc gttatgccca
     2581 ctatggtatt catcagcgtc cttttgtgat cgtcaaggcg gatgcgggta cttacggtat
     2641 gggtattatg accgcctatt cgggtgagga attccttgac ctcaaccgta aggaacgcac
     2701 ccggatggcg aagagtaaag aggggttgcc ggtcagtgac gtctttattc aagagggtgt
     2761 atataccttc gaacagaccg cgcaggaggc agtcgcagag ccggtggtct atatgatcgg
     2821 tcagcaggtg ctgggagggt tttaccgggt gcatacggag cgtggacggg atgagaatct
     2881 caacgctccg ggtgcgcatt ttgagccaat ggcctttggc cagacctgcg tgctcccttg
     2941 ccggaagtcg ccaccggatg cgccggccaa tcgttactat gcctatggcg tcatcgcgcg
     3001 cctcactctg gtggcggcgg cccgggaaat ggcagactgg cggcggcaag gtacactgga
     3061 gacaggccaa ggtacccatc atcaccatca ccactgacaa ataaaacgaa aggctcagtc
     3121 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     3181 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     3241 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     3301 ttttctagaa agcttgcggc cgcgatcaag caggtgcgac agacgtcata ctagactagt
     3361 actaactgtc acgaacccct gcaataactg tcacgccccc ctgcaataac tgtcacgaac
     3421 ccctgcaata actgtcacgc ccccaaacct gcaaacccag caggggcggg ggctggcggg
     3481 gtgttggaaa aatccatcca tgattatcta agaataatcc actaggcgcg gttatcagcg
     3541 cccttgtggg gcgctgctgc ccttgcccaa tatgcccggc cagaggccgg atagctggtc
     3601 tattcgctgc gctaggctac acaccgcccc accgctgcgc ggcaggggga aaggcgggca
     3661 aagcccgcta aaccccacac caaaccccgc agaaatacgc tggagcgctt ttagccgctt
     3721 tagcggcctt tccccctacc cgaagggtgg gggcgcgtgt gcagccccgc agggcctgtc
     3781 tcggtcgatc attcagcccg gctcatcctt ctggcgtggc ggcagaccga acaaggcgcg
     3841 gtcgtggtcg cgttcaaggt acgcatccat tgccgccatg agccgatcct ccggccactc
     3901 gctgctgttc accttggcca aaatcatggc ccccaccagc accttgcgcc ttgtttcgtt
     3961 cttgcgctct tgctgctgtt cccttgcccg cacccgctga atttcggcat tgattcgcgc
     4021 tcgttgttct tcgagcttgg ccagccgatc cgccgccttg ttgctcccct taaccatctt
     4081 gacaccccat tgttaatgtg ctgtctcgta ggctatcatg gaggcacagc ggcggcaatc
     4141 ccgaccctac tttgtagggg agggcgcact taccggtttc tcttcgagaa actggcctaa
     4201 cggccaccct tcgggcggtg cgctctccga gggccattgc atggagccga aaagcaaaag
     4261 caacagcgag gcagcatggc gatttatcac cttacggcga aaaccggcag caggtcgggc
     4321 ggccaatcgg ccagggccaa ggccgactac atccagcgcg aaggcaagta tgcccgcgac
     4381 atggatgaag tcttgcacgc cgaatccggg cacatgccgg agttcgtcga gcggcccgcc
     4441 gactactggg atgctgccga cctgtatgaa cgcgccaatg ggcggctgtt caaggaggtc
     4501 gaatttgccc tgccggtcga gctgaccctc gaccagcaga aggcgctggc gtccgagttc
     4561 gcccagcacc tgaccggtgc cgagcgcctg ccgtatacgc tggccatcca tgccggtggc
     4621 ggcgagaacc cgcactgcca cctgatgatc tccgagcgga tcaatgacgg catcgagcgg
     4681 cccgccgctc agtggttcaa gcggtacaac ggcaagaccc cggagaaggg cggggcacag
     4741 aagaccgaag cgctcaagcc caaggcatgg cttgagcaga cccgcgaggc atgggccgac
     4801 catgccaacc gggcattaga gcgggctggc cacgacgccc gcattgacca cagaacactt
     4861 gaggcgcagg gcatcgagcg cctgcccggt gttcacctgg ggccgaacgt ggtggagatg
     4921 gaaggccggg gcatccgcac cgaccgggca gacgtggccc tgaacatcga caccgccaac
     4981 gcccagatca tcgacttaca ggaataccgg gaggcaatag accatgaacg caatcgacag
     5041 agtgaagaaa tccagaggca tcaacgagtt agcggagcag atcgaaccgc tggcccagag
     5101 catggcgaca ctggccgacg aagcccggca ggtcatgagc cagacccagc aggccagcga
     5161 ggcgcaggcg gcggagtggc tgaaagccca gcgccagaca ggggcggcat gggtggagct
     5221 ggccaaagag ttgcgggagg tagccgccga ggtgagcagc gccgcgcaga gcgcccggag
     5281 cgcgtcgcgg gggtggcact ggaagctatg gctaaccgtg atgctggctt ccatgatgcc
     5341 tacggtggtg ctgctgatcg catcgttgct cttgctcgac ctgacgccac tgacaaccga
     5401 ggacggctcg atctggctgc gcttggtggc ccgatgaaga acgacaggac tttgcaggcc
     5461 ataggccgac agctcaaggc catgggctgt gagcgcttcg atatcggcgt cagggacgcc
     5521 accaccggcc agatgatgaa ccgggaatgg tcagccgccg aagtgctcca gaacacgcca
     5581 tggctcaagc ggatgaatgc ccagggcaat gacgtgtata tcaggcccgc cgagcaggag
     5641 cggcatggtc tggtgctggt ggacgacctc agcgagtttg acctggatga catgaaagcc
     5701 gagggccggg agcctgccct ggtagtggaa accagcccga agaactatca ggcatgggtc
     5761 aaggtggccg acgccgcagg cggtgaactt cgggggcaga ttgcccggac gctggccagc
     5821 gagtacgacg ccgacccggc cagcgccgac agccgccact atggccgctt ggcgggcttc
     5881 accaaccgca aggacaagca caccacccgc gccggttatc agccgtgggt gctgctgcgt
     5941 gaatccaagg gcaagaccgc caccgctggc ccggcgctgg tgcagcaggc tggccagcag
     6001 atcgagcagg cccagcggca gcaggagaag gcccgcaggc tggccagcct cgaactgccc
     6061 gagcggcagc ttagccgcca ccggcgcacg gcgctggacg agtaccgcag cgagatggcc
     6121 gggctggtca agcgcttcgg tgatgacctc agcaagtgcg actttatcgc cgcgcagaag
     6181 ctggccagcc ggggccgcag tgccgaggaa atcggcaagg ccatggccga ggccagccca
     6241 gcgctggcag agcgcaagcc cggccacgaa gcggattaca tcgagcgcac cgtcagcaag
     6301 gtcatgggtc tgcccagcgt ccagcttgcg cgggccgagc tggcacgggc accggcaccc
     6361 cgccagcgag gcatggacag gggcgggcca gatttcagca tgtagtgctt gcgttggtac
     6421 tcacgcctgt tatactatga gtactcacgc acagaagggg gttttatgga atacgaaaaa
     6481 agcgcttcag ggtcggtcta cctgatcaaa agtgacaagg gctattggtt gcccggtggc
     6541 tttggttata cgtcaaacaa ggccgaggct ggccgctttt cagtcgctga tatggccagc
     6601 cttaaccttg acggctgcac cttgtccttg ttccgcgaag acaagccttt cggccccggc
     6661 aagtttctcg gtgactgata tgaaagacca aaaggacaag cagaccggcg acctgctggc
     6721 cagccctgac gctgtacgcc aagcgcgata tgccgagcgc atgaaggcca aagggatgcg
     6781 tcagcgcaag ttctggctga ccgacgacga atacgaggcg ctgcgcgagt gcctggaaga
     6841 actcagagcg gcgcagggcg ggggtagtga ccccgccagc gcctaaccac caactgcctg
     6901 caaaggaggc aatcaatggc tacccataag cctatcaata ttctggaggc gttcgcagca
     6961 gcgccgccac cgctggacta cgttttgccc aacatggtgg ccggtacggt cggggcgctg
     7021 gtgtcgcccg gtggtgccgg taaatccatg ctggccctgc aactggccgc acagattgca
     7081 ggcgggccgg atctgctgga ggtgggcgaa ctgcccaccg gcccggtgat ctacctgccc
     7141 gccgaagacc cgcccaccgc cattcatcac cgcctgcacg cccttggggc gcacctcagc
     7201 gccgaggaac ggcaagccgt ggctgacggc ctgctgatcc agccgctgat cggcagcctg
     7261 cccaacatca tggccccgga gtggttcgac ggcctcaagc gcgccgccga gggccgccgc
     7321 ctgatggtgc tggacacgct gcgccggttc cacatcgagg aagaaaacgc cagcggcccc
     7381 atggcccagg tcatcggtcg catggaggcc atcgccgccg ataccgggtg ctctatcgtg
     7441 ttcctgcacc atgccagcaa gggcgcggcc atgatgggcg caggcgacca gcagcaggcc
     7501 agccggggca gctcggtact ggtcgataac atccgctggc agtcctacct gtcgagcatg
     7561 accagcgccg aggccgagga atggggtgtg gacgacgacc agcgccggtt cttcgtccgc
     7621 ttcggtgtga gcaaggccaa ctatggcgca ccgttcgctg atcggtggtt caggcggcat
     7681 gacggcgggg tgctcaagcc cgccgtgctg gagaggcagc gcaagagcaa gggggtgccc
     7741 cgtggtgaag cctaagaaca agcacagcct cagccacgtc cggcacgacc cggcgcactg
     7801 tctggccccc ggcctgttcc gtgccctcaa gcggggcgag cgcaagcgca gcaagctgga
     7861 cgtgacgtat gactacggcg acggcaagcg gatcgagttc agcggcccgg agccgctggg
     7921 cgctgatgat ctgcgcatcc tgcaagggct ggtggccatg gctgggccta atggcctagt
     7981 gcttggcccg gaacccaaga ccgaaggcgg acggcagctc cggctgttcc tggaacccaa
     8041 gtgggaggcc gtcaccgctg atgccatggt ggtcaaaggt agctatcggg cgctggcaaa
     8101 ggaaatcggg gcagaggtcg atagtggtgg ggcgctcaag cacatacagg actgcatcga
     8161 gcgcctttgg aaggtatcca tcatcgccca gaatggccgc aagcggcagg ggtttcggct
     8221 gctgtcggag tacgccagcg acgaggcgga cgggcgcctg tacgtggccc tgaacccctt
     8281 gatcgcgcag gccgtcatgg gtggcggcca gcatgtgcgc atcagcatgg acgaggtgcg
     8341 ggcgctggac agcgaaaccg cccgcctgct gcaccagcgg ctgtgtggct ggatcgaccc
     8401 cggcaaaacc ggcaaggctt ccatagatac cttgtgcggc tatgtctggc cgtcagaggc
     8461 cagtggttcg accatgcgca agcgccgcca gcgggtgcgc gaggcgttgc cggagctggt
     8521 cgcgctgggc tggacggtaa ccgagttcgc ggcgggcaag tacgacatca cccggcccaa
     8581 ggcggcaggc tgaccccccc cactctattg taaacaagac atttttatct tttatattca
     8641 atggcttatt ttcctgctaa ttggtaatac catgaaaaat accatgctca gaaaaggctt
     8701 aacaatattt tgaaaaattg cctactgagc gctgccgcac agctccatag gccgctttcc
     8761 tggcttt
//
