LOCUS       sequence-363051-        8230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic conjugative shuttle vector for Escherichia coli and 
            Methanothermobacter thermautotrophicus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8230)
  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 8230)
  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: pMVS-V1
FEATURES             Location/Qualifiers
     source          1..8230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(750..761)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3908..3917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4462..4478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4462..4478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(4675..5046)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5079..5188
                     /note="incP origin of transfer"
     primer_bind     complement(5338..5355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5509..6097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5589..5608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7071..7865
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 cgcgccgcaa aaaaatgcaa ataaaatttg gggtggcagg gtggagtaaa agtgtagtgt
       61 gatgatagtg gattccatct acctggggtg tcatccccca ctatggggtt gtgtttttct
      121 gtggttttgt gtgtgtttct gtggttgtag tgacacttta cttcggtata tcattatatt
      181 acgaagtcaa atcatggtac tgtgccgatg ttatggggtt aattgtttat actgtgacac
      241 accatagtta tatgatatga tcataggagt ggattgtatg gcgatggagg atgttggcgt
      301 gccattcagt aacaggttaa ccaggcaggg gaacattaag gtgcctgctg acttgaggga
      361 tgctttgaag ttgaagcctg gtgacctcct ggtggtggag atcaaaaaag ttgacaggtc
      421 atagtgacac tctcccctat tggcgaggca ttttaactct ttttatgctt ttacggtttt
      481 gcagaaaatc gcattccctg taagataata taagaaaaaa taatatagta tattaactgt
      541 tttcatggcc cgggctaagt tttaatagca agaacaacaa gaaatagtta tgaaaaagcg
      601 ttgttgagtg gggtggaaat accgcacacc aaaaccccac caacacagtc accctgcaga
      661 aaatgtaggg gtacacacct tgcagaatgg taccctgcat aagggttgta aagccctggc
      721 ttattctatg actgccaggg catataaaat cttccctcga ttctgcacct tgattctgca
      781 gggtgacacc aaaaacaaaa ggggtgatca ccttggataa aaaggcgata acagtcgaga
      841 taggtataga ggaatacttg aaactgcagg atttaagggt tgaatccctc aagactggta
      901 ggtttgtgtc tgttgggtgt gtcatgaggg atgcccttaa acactacctg caggaggttt
      961 acctgagaga gaaatacctc cagcaggcag aggacaataa gaaggagttg ataagatgag
     1021 tggcaaccac aaccacccac cccacagaac cacccctata gaggtgatgt gcaatccacc
     1081 cacaaaggag gtgaactaca atgaatagta tgtcaaagaa ggaatataaa cattttgagc
     1141 gatcaataac cctacacatt aaacacgctg attttgaatt agtaggccag aaatacaggg
     1201 gcagagacct ctacgagtac ttgttcatca aggggagtaa accaatccac acagccacag
     1261 gtaaaaccag caacctctac gagatcatat cagaggataa aggccctgat gaagccctca
     1321 aaatcatagg ggacacattc acagaggatg atataaacat cctcctcagg ggagggttcc
     1381 atgatgataa caagacccct gagggtgtcc tcgagttaat ccagcacatc ctcctcgcag
     1441 gtgaggtgct ccacccagga ggggatgtta ttgaacccca atcattcaag gactaccctg
     1501 agaagataca agcctacgct gaccagttaa tcaatgatga cagcatcgac atcctcgaat
     1561 ccatcaccag agtcataggg gaggcccatt atggtgatga gaaggccgtg aaattactcc
     1621 tattatccat cggcacccta ttcctcaggg acaccccacc agtccaccag gcactgaggg
     1681 gctccacagg ttcaggtaaa acagaccttg tattgaagac agtactcgca gtcccagaga
     1741 ggtatgtgca catcctcaga tcagcatcac ctaaatactt attctacgca tcagagactg
     1801 gcatcctccg tgaagactac aatatcttcg tatttgatga tattgagttg aatgatgaga
     1861 tcatagcgat ctccaagacc atcacagata acatcctccc agagaaggaa caccacaccg
     1921 tgaaggatca ggaggccctg aaactggaga tcccaggtga gggcctggcg atattcacaa
     1981 gggcgaggga catccatgat aatgaactca atgacaggct actctacaat aacccagtgg
     2041 aggatgagga tcacagcaga tttgtgaagg agaagatcaa agaggaggcc atctcagggt
     2101 cagtgatgga tgataagagg atcatggagg cctacgaggt catcagggct gtctatgagc
     2161 gattgataga gggtgatgtc agggtttaca acccctacct ccacctactc gacctgagag
     2221 ggtatagtaa cagggacatc aagcacatag ttggacttgt gaaggccgtg acatggtaca
     2281 ggcaacataa aaggaggagg gaggggagtt atgtccgtga aggtggccat gacattatca
     2341 tcggcactga ggatgacctg agggatgcat tggagttgtg gatgtccata gacaccctcc
     2401 aaaggtataa gttggacagg aaacaggaga aattcctcaa atccctacca gaatactctg
     2461 atgagttata taaacagcat actaattcaa tcttttggga tgatcctgaa cttccaactt
     2521 atagtggtct cgctgagagg cttgggttga gtaaatcaac aatatatcgg tgggttaatg
     2581 ggaggcatga gaaagacaga gacatcgagg gcttaaagga taaggggaat aagggtcttg
     2641 tcattatcaa accactggaa cctgagaatc ctaaatcacc ctcattgata taccgcaaac
     2701 caggtattgt tgggaaacac ctccaactca aaaatggcgg agaagcctca gaatctggat
     2761 ttaccctgtt ccccagtgtt gacatggact tgggaacagc ggaaaatgct cggatttgga
     2821 aaaaaaccat attccatagt atggttttag gtgtttccca aattcaaagt aacatacaag
     2881 acaatttaat acaagaatgg cttgataata atactaatcc catggagaca catgaggatg
     2941 ttgttgaatt cattgtcgat gtcagggcct tcatccatga caacctgcag acacctcctg
     3001 atgtgacctg caacccagta tataccatac tgggaaatgg gaaacacccc ctggaaacag
     3061 gtggagaatc ctccaatcac ggagataagc atgatttggt gttgaccagg gccgggtcaa
     3121 tagtggggaa tagggtaaac agtgaaaata agggctctga gccctccgat gaaatgacat
     3181 gtttccccca aaaaaattat tactctggtg tgggtggtga tctgcaggct gagtacactg
     3241 atgcaatggc acggggtgat gactcgatga taggtgaact tgaggcgagg atcccaccag
     3301 acacacctga gaaccttgag gagaggaggg tcaccctgaa aaagaaactc aaggataaga
     3361 gtatcagtgc cggggagaag atggaactcc tcaaggaact gggaggggtg ctctcaactc
     3421 taatgaggaa accagtggat aatgagagga ttgtcgagta tgcgaaggaa ttgggtgagg
     3481 ttcggcggga gatagaggat atgcaggggg ggtgatccat tatgcccctc tccttagcaa
     3541 cttcaaaccg tgtccctgag gcttggaggg atctgatcat atcggctatg agtaacctcg
     3601 cacccctcgg gagggatccg aatacaaagg aattcacaga tgaactccac cacctctcgg
     3661 gggaagatat ggggactgtg atggaaatcc tggggattgc agtggctgag gggttcctgg
     3721 aatacgatga tcattacagg ctcatcctcg gccctgcagg tgaggagtat gttcggctgt
     3781 catcacgggt taaggttgag gagacaagga gattgacact tcgaagggcc ctcaggagat
     3841 tcagggataa aaggagggcc tgcaggcacc tgtcccttgg ggataaggcc ctttatgatt
     3901 actattacca tggtggcctg tgaattataa ttcaatgtcc ttgttgggtg gaatgtggaa
     3961 gtcaacaaaa ccctcaagat ccaaatcacc aaagccttca agtaacccct taaggtcttc
     4021 aggtttgaac tctgggatgt catcaaggtt aaactctaag agttccttaa ggtcggggca
     4081 ttcaaggtca tcaaatactg ggaggtcttc aaggcaaaag tctattagtg gttcattagg
     4141 gtcaaatcct attagttctt ccaggttgaa ctctaaaacc ctgttatcat caatcttcat
     4201 gtgtaaatat ctatatcctc catggattaa taattttggg aattttggga gggttgaacc
     4261 accacccttg atagaaaaaa atctaaaaaa atctaaaaaa atcaaaataa aaattcaccc
     4321 ctggactggg ttcccctata atcaagtcct gtctgagaaa gcaaaaaaat acaagaaaat
     4381 ccgtggatga ctccattatg aatctatttt gaatccattg cgttgcgctg cgatcgcggt
     4441 catatggata cagcggccgc ggtcatagct gtttcctgat aaaaaaattg tagataaaac
     4501 tattttataa aatttatcta caattttttt atcctgcagg gggcccgatc ggtcttgcct
     4561 tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg
     4621 gggacgtgct tggcaatcac gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg
     4681 ctctgccctc gggcggacca cgcccatcat gaccttgcca agctcgtcct gcttctcttc
     4741 gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc
     4801 ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag
     4861 ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag
     4921 caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc
     4981 gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc
     5041 agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg
     5101 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
     5161 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
     5221 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
     5281 aatcgctata atgaccccga agcagggggc cctgcgtatt gggcgctctt ccgcttcctc
     5341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6181 aaaaaggagt ttaaacttag ggtaacaaaa aacaccgtat ttctacgatg tttttgctta
     6241 aatacttgtt tttagttaca gacaaacctg aagttaacta tttatcaatt cctgcaattc
     6301 gtttacaaaa cggcaaatgt gaaatccgtc acatactgcg tgatgaactt gaattgccaa
     6361 aggaagtata attttgttat cttctttata atatttcccc atagtaaaaa taggaatcaa
     6421 ataatcatat cctttctgca aattcagatt aaagccatcg aaggttgacc acggtatcat
     6481 agatacatta aaaatgtttt ccggagcatt tggctttcct tccattctat gattgtttcc
     6541 ataccgttgc gtatcacttt cataatctgc taaaaatgat ttaaagtcag acttacactc
     6601 agtccaaagg ctggaaaatg tttcagtatc attgtgaaat attgtatagc ttggtatcat
     6661 ctcatcatat atccccaatt caccatcttg attgattgcc gtcctaaact ctgaatggcg
     6721 gtttacaatc attgcaatat aataaagcat tgcaggatat agtttcattc ccttttcctt
     6781 tatttgtgtg atatccactt taacggtcat gctgtaggta caaggtacac ttgcaaagta
     6841 gtggtcaaaa tactcttttc tgttccaact atttttatca attttttcaa ataccatcta
     6901 agttccctct caaattcaag tttatcgctc taatgaacaa agatattata ccacattttt
     6961 gtgaattttt caacttgccc actggccggc cgcccatttt ttgagtattc tttatatagg
     7021 gatatagtaa tagataatat cacggtaacc accggcagca ggtgatgcat atggctaaaa
     7081 tgagaatatc accggaattg aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg
     7141 aaggaatgtc tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa
     7201 aaatgacgga cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga
     7261 tgctatggct ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg
     7321 gctggagcaa tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag
     7381 atgaacaaag ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact
     7441 ccatcgacat atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg
     7501 attacttact gaataacgat ctggccgatg tggattgcga aaactgggaa gaagacactc
     7561 catttaaaga tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac
     7621 ttgtcttttc ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa
     7681 gtggctttat tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct
     7741 gcgtccggtc gatcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact
     7801 tactggggat caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt
     7861 tttagcgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct
     7921 ccgaccgaag ccacccgggg cggccccgcc gaccccgcac ccgcccccga ggcccaccgc
     7981 gggggacaca ccgaacacgc cgaccctgct gaacacgcgg cgcagttcgg tgcccaggag
     8041 cggatcggga attaattcga agctgctggt gaaagagacc ctatcttacc tgctaaaatc
     8101 taagttaatt actaatttat tattaattta ttattagatt gggcaaaata gtaaaagaaa
     8161 actaaaggaa acctaatatg gtttcctttt tttatatatt tttaattcac tgggggcaat
     8221 tctgtcaggg
//
