LOCUS       62056_23135        8395 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Cloning vector pYI46, complete sequence.
ACCESSION   MZ420733
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  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 8395)
  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 8395)
  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..8395
                     /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             2705..2722
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2726..2812
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2904..2931
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      complement(3000..3394)
                     /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            3736..3823
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             5062..6030
                     /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             6544..7380
                     /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             7370..8218
                     /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 ccatgagtgc gctcaaggcc gctattctta tggatcccat
     1801 taccgggatc aaaccggcca aagatacgac gttcgccatg ctgctggcgg cgcaggcgcg
     1861 tgggcatcaa tgccgggtgt ttggcctgcc ggacctgttt tttcgggatg ggcgttcctg
     1921 gggacgactc cgtaccatca tcgtacgtga tgatgtggaa gattatttta cgctgggtga
     1981 gacagaagag cgccccctcg acgagatgga tgtggtcctc atgcgtaagg accctcctgt
     2041 gggtctggaa tatctcaccg cctgctatct gctggagcac gccggcacct gggtggtcaa
     2101 cgatccggtc agtctgcgca atgccaatga aaagctctat gccctgcatt tcccggaatt
     2161 tctgccgccc ttgctggtca gccgggatct cggggatttg cgggcctttc tcgccgaaca
     2221 cggcgaaatc gtcgtcaaac cgctctcggc gcgtggtggc gaaggggtgt tctatctgca
     2281 cctgcaggac cgcaatgtcg gcagtattct ggagacggtg acgggctggg gccagcatca
     2341 tgtgatggcg cagcgctacc tgcccgcgat tcgcgagggc gataaacgga ttctcctggt
     2401 ggatggtgtc ccggtgccag gtgccctgct gcgggtgcct tcggccgcgg attttcgcgg
     2461 taatctggtg gcaggtgcaa cgggggtagc ggcggagatc aatgaccgcg atcgggacat
     2521 ctgtgcggcg ataggcccgg ccttgcgcgc tgcgggcctg ctttttgtcg gcctggacgt
     2581 gatcggcggg tatgtgacgg agatcaatgt caccagcccc accggggcgc gggagatcag
     2641 gcgatttttt ggggtcgatg cggcggatct gctctggcag cgtctggagc gggggcgagg
     2701 tacccatcat caccatcacc actgacaaat aaaacgaaag gctcagtcga aagactgggc
     2761 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     2821 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     2881 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttctagaaag
     2941 cttgcggccg cgatcaagca ggtgcgacag acgtcatact agactagtac taactgtcac
     3001 gaacccctgc aataactgtc acgcccccct gcaataactg tcacgaaccc ctgcaataac
     3061 tgtcacgccc ccaaacctgc aaacccagca ggggcggggg ctggcggggt gttggaaaaa
     3121 tccatccatg attatctaag aataatccac taggcgcggt tatcagcgcc cttgtggggc
     3181 gctgctgccc ttgcccaata tgcccggcca gaggccggat agctggtcta ttcgctgcgc
     3241 taggctacac accgccccac cgctgcgcgg cagggggaaa ggcgggcaaa gcccgctaaa
     3301 ccccacacca aaccccgcag aaatacgctg gagcgctttt agccgcttta gcggcctttc
     3361 cccctacccg aagggtgggg gcgcgtgtgc agccccgcag ggcctgtctc ggtcgatcat
     3421 tcagcccggc tcatccttct ggcgtggcgg cagaccgaac aaggcgcggt cgtggtcgcg
     3481 ttcaaggtac gcatccattg ccgccatgag ccgatcctcc ggccactcgc tgctgttcac
     3541 cttggccaaa atcatggccc ccaccagcac cttgcgcctt gtttcgttct tgcgctcttg
     3601 ctgctgttcc cttgcccgca cccgctgaat ttcggcattg attcgcgctc gttgttcttc
     3661 gagcttggcc agccgatccg ccgccttgtt gctcccctta accatcttga caccccattg
     3721 ttaatgtgct gtctcgtagg ctatcatgga ggcacagcgg cggcaatccc gaccctactt
     3781 tgtaggggag ggcgcactta ccggtttctc ttcgagaaac tggcctaacg gccacccttc
     3841 gggcggtgcg ctctccgagg gccattgcat ggagccgaaa agcaaaagca acagcgaggc
     3901 agcatggcga tttatcacct tacggcgaaa accggcagca ggtcgggcgg ccaatcggcc
     3961 agggccaagg ccgactacat ccagcgcgaa ggcaagtatg cccgcgacat ggatgaagtc
     4021 ttgcacgccg aatccgggca catgccggag ttcgtcgagc ggcccgccga ctactgggat
     4081 gctgccgacc tgtatgaacg cgccaatggg cggctgttca aggaggtcga atttgccctg
     4141 ccggtcgagc tgaccctcga ccagcagaag gcgctggcgt ccgagttcgc ccagcacctg
     4201 accggtgccg agcgcctgcc gtatacgctg gccatccatg ccggtggcgg cgagaacccg
     4261 cactgccacc tgatgatctc cgagcggatc aatgacggca tcgagcggcc cgccgctcag
     4321 tggttcaagc ggtacaacgg caagaccccg gagaagggcg gggcacagaa gaccgaagcg
     4381 ctcaagccca aggcatggct tgagcagacc cgcgaggcat gggccgacca tgccaaccgg
     4441 gcattagagc gggctggcca cgacgcccgc attgaccaca gaacacttga ggcgcagggc
     4501 atcgagcgcc tgcccggtgt tcacctgggg ccgaacgtgg tggagatgga aggccggggc
     4561 atccgcaccg accgggcaga cgtggccctg aacatcgaca ccgccaacgc ccagatcatc
     4621 gacttacagg aataccggga ggcaatagac catgaacgca atcgacagag tgaagaaatc
     4681 cagaggcatc aacgagttag cggagcagat cgaaccgctg gcccagagca tggcgacact
     4741 ggccgacgaa gcccggcagg tcatgagcca gacccagcag gccagcgagg cgcaggcggc
     4801 ggagtggctg aaagcccagc gccagacagg ggcggcatgg gtggagctgg ccaaagagtt
     4861 gcgggaggta gccgccgagg tgagcagcgc cgcgcagagc gcccggagcg cgtcgcgggg
     4921 gtggcactgg aagctatggc taaccgtgat gctggcttcc atgatgccta cggtggtgct
     4981 gctgatcgca tcgttgctct tgctcgacct gacgccactg acaaccgagg acggctcgat
     5041 ctggctgcgc ttggtggccc gatgaagaac gacaggactt tgcaggccat aggccgacag
     5101 ctcaaggcca tgggctgtga gcgcttcgat atcggcgtca gggacgccac caccggccag
     5161 atgatgaacc gggaatggtc agccgccgaa gtgctccaga acacgccatg gctcaagcgg
     5221 atgaatgccc agggcaatga cgtgtatatc aggcccgccg agcaggagcg gcatggtctg
     5281 gtgctggtgg acgacctcag cgagtttgac ctggatgaca tgaaagccga gggccgggag
     5341 cctgccctgg tagtggaaac cagcccgaag aactatcagg catgggtcaa ggtggccgac
     5401 gccgcaggcg gtgaacttcg ggggcagatt gcccggacgc tggccagcga gtacgacgcc
     5461 gacccggcca gcgccgacag ccgccactat ggccgcttgg cgggcttcac caaccgcaag
     5521 gacaagcaca ccacccgcgc cggttatcag ccgtgggtgc tgctgcgtga atccaagggc
     5581 aagaccgcca ccgctggccc ggcgctggtg cagcaggctg gccagcagat cgagcaggcc
     5641 cagcggcagc aggagaaggc ccgcaggctg gccagcctcg aactgcccga gcggcagctt
     5701 agccgccacc ggcgcacggc gctggacgag taccgcagcg agatggccgg gctggtcaag
     5761 cgcttcggtg atgacctcag caagtgcgac tttatcgccg cgcagaagct ggccagccgg
     5821 ggccgcagtg ccgaggaaat cggcaaggcc atggccgagg ccagcccagc gctggcagag
     5881 cgcaagcccg gccacgaagc ggattacatc gagcgcaccg tcagcaaggt catgggtctg
     5941 cccagcgtcc agcttgcgcg ggccgagctg gcacgggcac cggcaccccg ccagcgaggc
     6001 atggacaggg gcgggccaga tttcagcatg tagtgcttgc gttggtactc acgcctgtta
     6061 tactatgagt actcacgcac agaagggggt tttatggaat acgaaaaaag cgcttcaggg
     6121 tcggtctacc tgatcaaaag tgacaagggc tattggttgc ccggtggctt tggttatacg
     6181 tcaaacaagg ccgaggctgg ccgcttttca gtcgctgata tggccagcct taaccttgac
     6241 ggctgcacct tgtccttgtt ccgcgaagac aagcctttcg gccccggcaa gtttctcggt
     6301 gactgatatg aaagaccaaa aggacaagca gaccggcgac ctgctggcca gccctgacgc
     6361 tgtacgccaa gcgcgatatg ccgagcgcat gaaggccaaa gggatgcgtc agcgcaagtt
     6421 ctggctgacc gacgacgaat acgaggcgct gcgcgagtgc ctggaagaac tcagagcggc
     6481 gcagggcggg ggtagtgacc ccgccagcgc ctaaccacca actgcctgca aaggaggcaa
     6541 tcaatggcta cccataagcc tatcaatatt ctggaggcgt tcgcagcagc gccgccaccg
     6601 ctggactacg ttttgcccaa catggtggcc ggtacggtcg gggcgctggt gtcgcccggt
     6661 ggtgccggta aatccatgct ggccctgcaa ctggccgcac agattgcagg cgggccggat
     6721 ctgctggagg tgggcgaact gcccaccggc ccggtgatct acctgcccgc cgaagacccg
     6781 cccaccgcca ttcatcaccg cctgcacgcc cttggggcgc acctcagcgc cgaggaacgg
     6841 caagccgtgg ctgacggcct gctgatccag ccgctgatcg gcagcctgcc caacatcatg
     6901 gccccggagt ggttcgacgg cctcaagcgc gccgccgagg gccgccgcct gatggtgctg
     6961 gacacgctgc gccggttcca catcgaggaa gaaaacgcca gcggccccat ggcccaggtc
     7021 atcggtcgca tggaggccat cgccgccgat accgggtgct ctatcgtgtt cctgcaccat
     7081 gccagcaagg gcgcggccat gatgggcgca ggcgaccagc agcaggccag ccggggcagc
     7141 tcggtactgg tcgataacat ccgctggcag tcctacctgt cgagcatgac cagcgccgag
     7201 gccgaggaat ggggtgtgga cgacgaccag cgccggttct tcgtccgctt cggtgtgagc
     7261 aaggccaact atggcgcacc gttcgctgat cggtggttca ggcggcatga cggcggggtg
     7321 ctcaagcccg ccgtgctgga gaggcagcgc aagagcaagg gggtgccccg tggtgaagcc
     7381 taagaacaag cacagcctca gccacgtccg gcacgacccg gcgcactgtc tggcccccgg
     7441 cctgttccgt gccctcaagc ggggcgagcg caagcgcagc aagctggacg tgacgtatga
     7501 ctacggcgac ggcaagcgga tcgagttcag cggcccggag ccgctgggcg ctgatgatct
     7561 gcgcatcctg caagggctgg tggccatggc tgggcctaat ggcctagtgc ttggcccgga
     7621 acccaagacc gaaggcggac ggcagctccg gctgttcctg gaacccaagt gggaggccgt
     7681 caccgctgat gccatggtgg tcaaaggtag ctatcgggcg ctggcaaagg aaatcggggc
     7741 agaggtcgat agtggtgggg cgctcaagca catacaggac tgcatcgagc gcctttggaa
     7801 ggtatccatc atcgcccaga atggccgcaa gcggcagggg tttcggctgc tgtcggagta
     7861 cgccagcgac gaggcggacg ggcgcctgta cgtggccctg aaccccttga tcgcgcaggc
     7921 cgtcatgggt ggcggccagc atgtgcgcat cagcatggac gaggtgcggg cgctggacag
     7981 cgaaaccgcc cgcctgctgc accagcggct gtgtggctgg atcgaccccg gcaaaaccgg
     8041 caaggcttcc atagatacct tgtgcggcta tgtctggccg tcagaggcca gtggttcgac
     8101 catgcgcaag cgccgccagc gggtgcgcga ggcgttgccg gagctggtcg cgctgggctg
     8161 gacggtaacc gagttcgcgg cgggcaagta cgacatcacc cggcccaagg cggcaggctg
     8221 acccccccca ctctattgta aacaagacat ttttatcttt tatattcaat ggcttatttt
     8281 cctgctaatt ggtaatacca tgaaaaatac catgctcaga aaaggcttaa caatattttg
     8341 aaaaattgcc tactgagcgc tgccgcacag ctccataggc cgctttcctg gcttt
//
