LOCUS       sequence-431894-       10426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conjugative shuttle vector for Escherichia coli and 
            Methanothermobacter thermautotrophicus encoding beta-galactosidase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10426)
  AUTHORS   Fink C, Beblawy S, Enkerlin AM, Muhling L, Angenent LT, Molitor B
  TITLE     A Shuttle-Vector System Allows Heterologous Gene Expression in the 
            Thermophilic Methanogen Methanothermobacter thermautotrophicus 
            DeltaH.
  JOURNAL   mBio. 2021 Dec 21;12(6):e0276621. doi: 10.1128/mBio.02766-21. Epub 
            2021 Nov 23.
  PUBMED    34809461
REFERENCE   2  (bases 1 to 10426)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "mBio."; 
            date: "2021-12-21"; volume: "12(6):e0276621. doi"; pages: " 
            10.1128/mBio.02766-21. Epub 2021 Nov 23"
COMMENT     Sequence Label: pMVS1111A:PhmtB-bgaB
FEATURES             Location/Qualifiers
     source          1..10426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             879..1673
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(4984..4995)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      8142..8151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(8696..8712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8696..8712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(8909..9280)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9313..9422
                     /note="incP origin of transfer"
     primer_bind     complement(9572..9589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9743..10331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9823..9842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaacttaggg taacaaaaaa caccgtattt ctacgatgtt tttgcttaaa tacttgtttt
       61 tagttacaga caaacctgaa gttaactatt tatcaattcc tgcaattcgt ttacaaaacg
      121 gcaaatgtga aatccgtcac atactgcgtg atgaacttga attgccaaag gaagtataat
      181 tttgttatct tctttataat atttccccat agtaaaaata ggaatcaaat aatcatatcc
      241 tttctgcaaa ttcagattaa agccatcgaa ggttgaccac ggtatcatag atacattaaa
      301 aatgttttcc ggagcatttg gctttccttc cattctatga ttgtttccat accgttgcgt
      361 atcactttca taatctgcta aaaatgattt aaagtcagac ttacactcag tccaaaggct
      421 ggaaaatgtt tcagtatcat tgtgaaatat tgtatagctt ggtatcatct catcatatat
      481 ccccaattca ccatcttgat tgattgccgt cctaaactct gaatggcggt ttacaatcat
      541 tgcaatataa taaagcattg caggatatag tttcattccc ttttccttta tttgtgtgat
      601 atccacttta acggtcatgc tgtaggtaca aggtacactt gcaaagtagt ggtcaaaata
      661 ctcttttctg ttccaactat ttttatcaat tttttcaaat accatctaag ttccctctca
      721 aattcaagtt tatcgctcta atgaacaaag atattatacc acatttttgt gaatttttca
      781 acttgcccac tggccggccg cccatttttt gagtattctt tatataggga tatagtaata
      841 gataatatca cggtaaccac cggcagcagg tgatgcatat ggctaaaatg agaatatcac
      901 cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa agatacggaa ggaatgtctc
      961 ctgctaaggt atataagctg gtgggagaaa atgaaaacct atatttaaaa atgacggaca
     1021 gccggtataa agggaccacc tatgatgtgg aacgggaaaa ggacatgatg ctatggctgg
     1081 aaggaaagct gcctgttcca aaggtcctgc actttgaacg gcatgatggc tggagcaatc
     1141 tgctcatgag tgaggccgat ggcgtccttt gctcggaaga gtatgaagat gaacaaagcc
     1201 ctgaaaagat tatcgagctg tatgcggagt gcatcaggct ctttcactcc atcgacatat
     1261 cggattgtcc ctatacgaat agcttagaca gccgcttagc cgaattggat tacttactga
     1321 ataacgatct ggccgatgtg gattgcgaaa actgggaaga agacactcca tttaaagatc
     1381 cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga agaggaactt gtcttttccc
     1441 acggcgacct gggagacagc aacatctttg tgaaagatgg caaagtaagt ggctttattg
     1501 atcttgggag aagcggcagg gcggacaagt ggtatgacat tgccttctgc gtccggtcga
     1561 tcagggagga tatcggggaa gaacagtatg tcgagctatt ttttgactta ctggggatca
     1621 agcctgattg ggagaaaata aaatattata ttttactgga tgaattgttt tagcgcccgc
     1681 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc
     1741 acccggggcg gccccgccga ccccgcaccc gcccccgagg cccaccgcgg gggacacacc
     1801 gaacacgccg accctgctga acacgcggcg cagttcggtg cccaggagcg gatcgggaat
     1861 taattcgaag ctgctggtga aagagaccct atcttacctg ctaaaatcta agttaattac
     1921 taatttatta ttaatttatt attagattgg gcaaaatagt aaaagaaaac taaaggaaac
     1981 ctaatatggt ttcctttttt tatatatttt taattcactg ggggcaattc tgtcagggcg
     2041 cgcctggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg ggcgctagac
     2101 cttgccggct tcgtcgtgtt ccctaaggac agcgacgtcg acgccctgaa tcctgagttc
     2161 accccccctg aagcatttgc catctatcat attctggtag atcttatctt ccggaaggga
     2221 gagtgtgacc tcatagtcgt tgtggttaat tataataagg tacttccatt catcggtctc
     2281 cctctgctga acttcgacat tctcagcaac ctccagtata ggatttatgt ggtgtttagc
     2341 aaacacctgt tcgagaagcc tgccaaggta gttgctgtca gggtatgttc ctacgtatat
     2401 gccctccccc tttccgtagc agttcctggt aacagcagga aggccggcat accaatcacc
     2461 tttgaatgtg gcgagaggct cagcaccttc cagccttatt atatcggccc atgtggtaca
     2521 gtcatactcg ccgtcgtttg agtagatctt attcaccttt gtctcgggat aaggaacgaa
     2581 ttcctccacg aagatgccga gaatgtccct cagcggtcct ggatatcccc cgaggtgcac
     2641 tctatcgttc tcatcgacta tcacactgaa aaagcttaca atcagggttc cgccgtttgc
     2701 gacaaactgc ctaaggttct catcttctcc ctctttcacc atatacagca tcggtgcaat
     2761 aacaacctta tattttgtga gatcgtcgga cggtcttaca aagtcgactg ctatgtttct
     2821 cttgtaaagc tctctataat atgcctctac tatgggaata tatctgagct tgttgtgcgg
     2881 tttggaactg agctcaactg cccaccagtt ttcccagtca aagataattg ccacctctgc
     2941 ctttattcta ctccccacga ggcagtcaag ttttttcagc tcctggccaa gctgggtaac
     3001 ttccctgtat attctattgt tttcgttaag aaagtggggc accattgctc cgtgaaactt
     3061 ctcagctcct gctctggact gcctccactg aaagaacatt atcccatcgg cacccctggc
     3121 gattgttgcg taactccaga gtctcataac ccccggcggc tttggcacat tgatatctct
     3181 ccaattaacg tgactggtga cctgctccat aagaatgaac ggctgcccct tcctaagtga
     3241 cctcatgagg tcattcatca ttgcgtgctg tatagggagt ccctccctgg gatctgggta
     3301 gctatcccag gtaacgatat ctacgtgctg agcccactga aagtagttga gtggcttgaa
     3361 tgatcccatg aaatttgtgg agaccgggat atcgggggtt acttccctga ggatctcctt
     3421 ttctgtaagg aagagtttga ggattgaatc attcatgaat ctgtagtaat caagctcctg
     3481 gctggggtta ataaatgttg gtgccttcct agggggatta atctcatccc agtggttata
     3541 tctctggccc cagaagtttg taccccatct ttcattaagt tcatcaatgg tcttatacct
     3601 ttctttaagc cattttctga aagcaactgc gcaattctca cagaaacact tacttacatg
     3661 gcaagcgtat tcgttattta cgtgccacat tttgagggct ggatgatttt tgtatctctc
     3721 agctatagcc cttaccagcc tctttatatg tgttataagc tgagggtgat ttgggcaata
     3781 atgctgtcta ctcccgaaac tcagtatcac accggactcg tcaataggga gtgaatcagg
     3841 gtatttcttc acgaaccagg cgggtgtggt tgcggtggcg gtccccagat ttatgtatac
     3901 cccatgatcg tagaggatgt ctatcacttt gtcgagccat tcaaaatcaa atacaccgtc
     3961 tgatggctcg attttggacc agctaaagat tccgagtgaa acaagattaa caccggcctt
     4021 ctgcataagt tttgcgtcct cgtaccatat ctcctcgggc cactgttctg ggttgtaatc
     4081 ccccccatag cagatggaac tgagaacgtt cataattaac cacctcattt tatgtgatat
     4141 tatctattca tataatccta tataaatata tcgctaattt taaggttttt ctgagccatc
     4201 ggttggttca tgggggcatt cctccaggct taattaaccc gcaaaaaaat gcaaataaaa
     4261 tttggggtgg cagggtggag taaaagtgta gtgtgatgat agtggattcc atctacctgg
     4321 ggtgtcatcc cccactatgg ggttgtgttt ttctgtggtt ttgtgtgtgt ttctgtggtt
     4381 gtagtgacac tttacttcgg tatatcatta tattacgaag tcaaatcatg gtactgtgcc
     4441 gatgttatgg ggttaattgt ttatactgtg acacaccata gttatatgat atgatcatag
     4501 gagtggattg tatggcgatg gaggatgttg gcgtgccatt cagtaacagg ttaaccaggc
     4561 aggggaacat taaggtgcct gctgacttga gggatgcttt gaagttgaag cctggtgacc
     4621 tcctggtggt ggagatcaaa aaagttgaca ggtcatagtg acactctccc ctattggcga
     4681 ggcattttaa ctctttttat gcttttacgg ttttgcagaa aatcgcattc cctgtaagat
     4741 aatataagaa aaaataatat agtatattaa ctgttttcat ggcccgggct aagttttaat
     4801 agcaagaaca acaagaaata gttatgaaaa agcgttgttg agtggggtgg aaataccgca
     4861 caccaaaacc ccaccaacac agtcaccctg cagaaaatgt aggggtacac accttgcaga
     4921 atggtaccct gcataagggt tgtaaagccc tggcttattc tatgactgcc agggcatata
     4981 aaatcttccc tcgattctgc accttgattc tgcagggtga caccaaaaac aaaaggggtg
     5041 atcaccttgg ataaaaaggc gataacagtc gagataggta tagaggaata cttgaaactg
     5101 caggatttaa gggttgaatc cctcaagact ggtaggtttg tgtctgttgg gtgtgtcatg
     5161 agggatgccc ttaaacacta cctgcaggag gtttacctga gagagaaata cctccagcag
     5221 gcagaggaca ataagaagga gttgataaga tgagtggcaa ccacaaccac ccaccccaca
     5281 gaaccacccc tatagaggtg atgtgcaatc cacccacaaa ggaggtgaac tacaatgaat
     5341 agtatgtcaa agaaggaata taaacatttt gagcgatcaa taaccctaca cattaaacac
     5401 gctgattttg aattagtagg ccagaaatac aggggcagag acctctacga gtacttgttc
     5461 atcaagggga gtaaaccaat ccacacagcc acaggtaaaa ccagcaacct ctacgagatc
     5521 atatcagagg ataaaggccc tgatgaagcc ctcaaaatca taggggacac attcacagag
     5581 gatgatataa acatcctcct caggggaggg ttccatgatg ataacaagac ccctgagggt
     5641 gtcctcgagt taatccagca catcctcctc gcaggtgagg tgctccaccc aggaggggat
     5701 gttattgaac cccaatcatt caaggactac cctgagaaga tacaagccta cgctgaccag
     5761 ttaatcaatg atgacagcat cgacatcctc gaatccatca ccagagtcat aggggaggcc
     5821 cattatggtg atgagaaggc cgtgaaatta ctcctattat ccatcggcac cctattcctc
     5881 agggacaccc caccagtcca ccaggcactg aggggctcca caggttcagg taaaacagac
     5941 cttgtattga agacagtact cgcagtccca gagaggtatg tgcacatcct cagatcagca
     6001 tcacctaaat acttattcta cgcatcagag actggcatcc tccgtgaaga ctacaatatc
     6061 ttcgtatttg atgatattga gttgaatgat gagatcatag cgatctccaa gaccatcaca
     6121 gataacatcc tcccagagaa ggaacaccac accgtgaagg atcaggaggc cctgaaactg
     6181 gagatcccag gtgagggcct ggcgatattc acaagggcga gggacatcca tgataatgaa
     6241 ctcaatgaca ggctactcta caataaccca gtggaggatg aggatcacag cagatttgtg
     6301 aaggagaaga tcaaagagga ggccatctca gggtcagtga tggatgataa gaggatcatg
     6361 gaggcctacg aggtcatcag ggctgtctat gagcgattga tagagggtga tgtcagggtt
     6421 tacaacccct acctccacct actcgacctg agagggtata gtaacaggga catcaagcac
     6481 atagttggac ttgtgaaggc cgtgacatgg tacaggcaac ataaaaggag gagggagggg
     6541 agttatgtcc gtgaaggtgg ccatgacatt atcatcggca ctgaggatga cctgagggat
     6601 gcattggagt tgtggatgtc catagacacc ctccaaaggt ataagttgga caggaaacag
     6661 gagaaattcc tcaaatccct accagaatac tctgatgagt tatataaaca gcatactaat
     6721 tcaatctttt gggatgatcc tgaacttcca acttatagtg gtctcgctga gaggcttggg
     6781 ttgagtaaat caacaatata tcggtgggtt aatgggaggc atgagaaaga cagagacatc
     6841 gagggcttaa aggataaggg gaataagggt cttgtcatta tcaaaccact ggaacctgag
     6901 aatcctaaat caccctcatt gatataccgc aaaccaggta ttgttgggaa acacctccaa
     6961 ctcaaaaatg gcggagaagc ctcagaatct ggatttaccc tgttccccag tgttgacatg
     7021 gacttgggaa cagcggaaaa tgctcggatt tggaaaaaaa ccatattcca tagtatggtt
     7081 ttaggtgttt cccaaattca aagtaacata caagacaatt taatacaaga atggcttgat
     7141 aataatacta atcccatgga gacacatgag gatgttgttg aattcattgt cgatgtcagg
     7201 gccttcatcc atgacaacct gcagacacct cctgatgtga cctgcaaccc agtatatacc
     7261 atactgggaa atgggaaaca ccccctggaa acaggtggag aatcctccaa tcacggagat
     7321 aagcatgatt tggtgttgac cagggccggg tcaatagtgg ggaatagggt aaacagtgaa
     7381 aataagggct ctgagccctc cgatgaaatg acatgtttcc cccaaaaaaa ttattactct
     7441 ggtgtgggtg gtgatctgca ggctgagtac actgatgcaa tggcacgggg tgatgactcg
     7501 atgataggtg aacttgaggc gaggatccca ccagacacac ctgagaacct tgaggagagg
     7561 agggtcaccc tgaaaaagaa actcaaggat aagagtatca gtgccgggga gaagatggaa
     7621 ctcctcaagg aactgggagg ggtgctctca actctaatga ggaaaccagt ggataatgag
     7681 aggattgtcg agtatgcgaa ggaattgggt gaggttcggc gggagataga ggatatgcag
     7741 ggggggtgat ccattatgcc cctctcctta gcaacttcaa accgtgtccc tgaggcttgg
     7801 agggatctga tcatatcggc tatgagtaac ctcgcacccc tcgggaggga tccgaataca
     7861 aaggaattca cagatgaact ccaccacctc tcgggggaag atatggggac tgtgatggaa
     7921 atcctgggga ttgcagtggc tgaggggttc ctggaatacg atgatcatta caggctcatc
     7981 ctcggccctg caggtgagga gtatgttcgg ctgtcatcac gggttaaggt tgaggagaca
     8041 aggagattga cacttcgaag ggccctcagg agattcaggg ataaaaggag ggcctgcagg
     8101 cacctgtccc ttggggataa ggccctttat gattactatt accatggtgg cctgtgaatt
     8161 ataattcaat gtccttgttg ggtggaatgt ggaagtcaac aaaaccctca agatccaaat
     8221 caccaaagcc ttcaagtaac cccttaaggt cttcaggttt gaactctggg atgtcatcaa
     8281 ggttaaactc taagagttcc ttaaggtcgg ggcattcaag gtcatcaaat actgggaggt
     8341 cttcaaggca aaagtctatt agtggttcat tagggtcaaa tcctattagt tcttccaggt
     8401 tgaactctaa aaccctgtta tcatcaatct tcatgtgtaa atatctatat cctccatgga
     8461 ttaataattt tgggaatttt gggagggttg aaccaccacc cttgatagaa aaaaatctaa
     8521 aaaaatctaa aaaaatcaaa ataaaaattc acccctggac tgggttcccc tataatcaag
     8581 tcctgtctga gaaagcaaaa aaatacaaga aaatccgtgg atgactccat tatgaatcta
     8641 ttttgaatcc attgcgttgc gctgcgatcg cggtcatatg gatacagcgg ccgcggtcat
     8701 agctgtttcc tgataaaaaa attgtagata aaactatttt ataaaattta tctacaattt
     8761 ttttatcctg cagggggccc gatcggtctt gccttgctcg tcggtgatgt acttcaccag
     8821 ctccgcgaag tcgctcttct tgatggagcg catggggacg tgcttggcaa tcacgcgcac
     8881 cccccggccg ttttagcggc taaaaaagtc atggctctgc cctcgggcgg accacgccca
     8941 tcatgacctt gccaagctcg tcctgcttct cttcgatctt cgccagcagg gcgaggatcg
     9001 tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc
     9061 ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg gacgtgctca tagtccacga
     9121 cgcccgtgat tttgtagccc tggccgacgg ccagcaggta ggccgacagg ctcatgccgg
     9181 ccgccgccgc cttttcctca atcgctcttc gttcgtctgg aaggcagtac accttgatag
     9241 gtgggctgcc cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta
     9301 cgccggcggt agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg
     9361 aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc
     9421 ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg
     9481 tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg
     9541 gggccctgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     9601 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     9661 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     9721 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     9781 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     9841 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     9901 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     9961 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    10021 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    10081 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    10141 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    10201 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    10261 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    10321 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    10381 actcacgtta agggattttg gtcatgagat tatcaaaaag gagttt
//
