LOCUS       sequence-379181-        9461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged ANME2bHR1 MCR in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9461)
  AUTHORS   Shao N, Fan Y, Chou CW, Yavari S, Williams RV, Amster IJ, Brown SM, 
            Drake IJ, Duin EC, Whitman WB, Liu Y
  TITLE     Expression of divergent methyl/alkyl coenzyme M reductases from 
            uncultured archaea.
  JOURNAL   Commun Biol. 2022 Oct 20;5(1):1113. doi: 10.1038/s42003-022-04057-6.
  PUBMED    36266535
REFERENCE   2  (bases 1 to 9461)
  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: "Commun 
            Biol."; date: "2022-10-20"; volume: "5(1):1113. doi"; pages: " 
            10.1038/s42003-022-04057-6"
COMMENT     Sequence Label: Ppst93-ANME2bHR1 MCR-NG
FEATURES             Location/Qualifiers
     source          1..9461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2561..2584
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2594..2617,2618..2653,2654..2677)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(6515..6534)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6634..6656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6694..6712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6780..6884
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6885..6953,6954..7745)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7103..7122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7916..8504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8405..8424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaactaaaa taccaaaata gaaacattta tatacatttg atgtgtaggg tataccataa
       61 acctgggagg tgtctcatat ggcagataaa attgatcttt actcagatag aggagcaaaa
      121 cttaaatcag gagttatttt agaagatatt tcaccattaa gaaacaaagc aattaaacaa
      181 attattcacg atacaaaaag atcagcagca gttgatttag caggaattga aaaagcactt
      241 gcatcaggaa aatacggagg aaaaggaaga caaattcagg gaagaaaact tgaacttgat
      301 gttgttaaaa atgcagattc aattaaagca aaaatggaag atcttgttag agttgataaa
      361 ggagatgata ctaacattga ttcattaaat ggaggaaaac aattatttgt tcaggttcca
      421 tcagcaagaa tggaagcagg agcagaattt gttgcatcat taacatgttc agcaatgtca
      481 tgtgttcagg caatgattga tacttttgat ctttcaatgt ttgttgttcc agaagttaaa
      541 gcagcaatta tgggacaata tccacagaca atggatatgg caggtggaaa cgttaaatca
      601 attttagaaa ttccacaaaa agatgaagga atgggacact cacttagaaa cattatggca
      661 aaccaccttg cagcagttac aaagaaaaat gcaatgaaca ctgcagcact ttcatcaatt
      721 tacgaacagg caggagtttt tgaaatggga aatgcattag gacaatttca gagacaccaa
      781 cttcttggac ttgcatacca gggacttaac gcagataacc ttgtttacga tgttacaaaa
      841 gcaaatggag caacaggaac tattggaact gttatgcact caattgttga aagaggatta
      901 gcagataaaa ttttagttgc agataaaaaa tttgattcag gatatgtttg ttacaaagca
      961 acagatgttg caaaatggaa tgcatataat gcagcaggaa ctcttgcatc aaaccttgtt
     1021 aactgtggag caggaagatc accacaacac gtttcatcaa cacttcttta ctacaacgat
     1081 cttgttgaaa aagaaactgg attaccagga tgtgattatg gaaaagttca gggaacagca
     1141 gttggatttt catttttctc acactcaatt tacggaggcg gaggaccagg agtttttaat
     1201 ggaaatcacg ttgttactag acactcaaga ggatttgcaa ttccatgtgt tgcagcagca
     1261 gtttcattag atgcaggagt tcaaatgttt acaccagaaa tgacttcagc attagttgga
     1321 gaaacatttg gatcatacga aacttttaga gcaccattaa aggcagtcgc cggagcagtg
     1381 taaataaaaa aaaagaggat ttaaatggtt gattattact cagcacctaa aaaaccaatt
     1441 caagttgaaa tttttccaca gaaacttctt atgccagata cagcagaaga acttcttaac
     1501 tcaattgata aagttactgg agttcttaga atgatgattc aaggatcagg attaccaatg
     1561 agagtttcat gtggaccagg aattggaaca ccaattgata gatcagatag aagaattatt
     1621 aacgtttcag gacaggcatt tgaacttaac attagagttg gaagaattag agttgaactt
     1681 gaagataaat cagcaaaaga accattaaga gaagcagttg aaaactcact tccaattcca
     1741 tttgaattta gagaaggatt tttctttaga aagaaagcat cacttactga ttacgcaaaa
     1801 tacggattta aatcaagaga agatgaatca attaacattg aagatgaaag acttcttgga
     1861 ttaattgacc ctaaagcaga cccaaaagac cgtatatgca tgttagaatc aaatgagtaa
     1921 tcatatgttt ggaagagaaa cacaatgtgt tgattgtaga gaatcaatgg gattaggaag
     1981 aggtggagga ttagcacaga gaggaacatt atcagaaact ggaagaccag atgttattgc
     2041 agttgcaatg tcaccaggaa gaagacacat tacaaaacca gtttgtgaaa ttacttatgg
     2101 acttagaaga gaaagaattc aagtttcagt tttagttctt aattcaggag caggagttcc
     2161 agaagattta cacaaagaat caactatggg atacggacca aaatttggac ttaacgcaaa
     2221 agaattagca cagattggaa gacacaaatt attagttctt caccttggaa acattgattc
     2281 acacgttatt tcaaaaacaa aagaagttct taaatatgtt gatattccaa ctattgttgc
     2341 atgtcaaaat ccaatggatt ttgaacagtt tgcagaagca ggagttagaa caagattagt
     2401 taaaccaaga ccagaaaaaa ttcaaactgg aggaactgtt atggaaattg tttcaggaat
     2461 tacaagagga gaaacttgtt caagagaaaa tttaaataag attgttaatt acgtaaagac
     2521 cattaatccg gaaaataact aaggagatga tgtaataatg gattacaaag atgatgatga
     2581 taaaggcgcc agctggtcac acccacaatt tgaaaaagga ggtggatcag gaggtggatc
     2641 aggaggttca gcatggtcac atcctcagtt tgaaaaatca gcaatggcat ataaaccaca
     2701 atattaccca ggatcaacat cagttgcaga aaatagaaga aaacacatgt caaacaacgt
     2761 tgaaaaatta agagaagttt cagatgaaga tttaacagca atgttaggac acagagcacc
     2821 aggatcagat tacccatcaa ctcacccacc attagcagaa atgggagaac cagattgtcc
     2881 agttagacaa atggttacac caactccagg aactgcagca ggagatagag ttagatatgt
     2941 tcagtgggca gattcaatgt acaatgcacc atcagttcca tattggagat cataccacgc
     3001 agcaattaat cacagaggag ttgatccagg aacattatca ggaagacaag ttgttgaagc
     3061 aagagaaaga gatatggaag catgtgcaaa agaacaggtt gaaacagaaa tgacttgtcc
     3121 aggattagca ggattaagag gagcaactgt tcacggacac tcatgtagat tacaggaaga
     3181 tggagttatg tttgatatgc ttgatagaag aagattagaa ggagatgtta ttattcaaga
     3241 taaagatcag gttggagttc cattagatag aaaagttaac atgggaaaac caatgtcaga
     3301 agcagaagca gcaaaaagaa caacttttta cagaattgat aatgttgcat ttagagatga
     3361 taaagaagtt attgaatgga tgcaaagagt ttgggaactg agaacattat atggatatca
     3421 gccccatgat taagaggtga tagaaaatga caaaacactt cgcattcgga actgaaaaga
     3481 ctgctatgga tcatttcaaa cttgcaatgg aaaacaaata cggaaaagaa tggggatcaa
     3541 accaacagaa cgatgatatt caagcaaaaa caactaaata tttaagatta ggatacgaac
     3601 agtcaccaag aaaagttgaa atggcaaaag caggagcaga tattgcaaag aaaagaggat
     3661 tacaagcata cgatccatta ttacacttag caggaattcc attaggacaa agacagttaa
     3721 caccatacac tcttggagga acagatattg ttgttgaagg agatgattta cactacgtta
     3781 acaacgcagc aatgcaacag atgtgggatg atattagaag aacatgtgtt gttggattag
     3841 atcttgcaca cgaaacatta gaaaaacgac ttggaaaaga agttacacca gaaactatta
     3901 actactactt agaagttctt aatcacgcat tacctggagg agcaattgtt caagaacaca
     3961 tggttgaaac tcacccagca atggttgatg attgttacgt taaagttttt acaggagatg
     4021 atactcttaa agatgaatta gattcacagt ttgttattga tattgataaa atgtttattc
     4081 cagatcacgc agcacaaatt aaagcatcaa ttggaaaagc atcatttcag gcagttcaca
     4141 ttccaacagt tgtttcaaga gttgcagatg gaggacaaac ttcaagatgg atggcaatgc
     4201 aggttggaat gtcatttatt tcagcatatc acatgtgtgc aggagaagca gcagttgcag
     4261 atttagcatt tacagcaaaa cacgcaggat tagttgaaat gtcagaaatg ttaccagcaa
     4321 gaagagcaag aggaccaaat gaacctggag gattatcatt tggacacatg tgtgatattg
     4381 ttcaaacatc aagaaaattt agagatgatc cagataaaat tgcattagaa acttgtgcag
     4441 cagcatcaat gttatacgat cagatttggc ttggaggata tatgtctgga ggagttggat
     4501 ttacaatgta tgcaactgca gcatacacaa acaacactgt tgatgatgat ctttacgcag
     4561 caacagaata cggatgggat acatacggaa ctgcagttgg aaaaactgtt gcaccatcaa
     4621 ttgatgttat taaagatatt ggaacatggg gaactattta cggattagaa ctttacgaaa
     4681 actacccaac atcattagaa gatcactttg gaggatcaca aagagcaact gttattgcaa
     4741 cagcaactgg agcagcatgt gcaattacaa ctggaaattc aaatgcagga ctttcagcat
     4801 ggtacctttg tatgtactta cacaaagaag cacacggaag attaggattt ttcggatacg
     4861 atttacaaga tcagtgtgga gcaactaacg tttactcata ccaatcagat gaaggacttc
     4921 ttggagaact tagaggagca aactacccaa actacgcaat gaatgttgga caccagggag
     4981 gatacacaac tgttttaaca ggagcatata ctggatgtgg acacggattt tcaacaaatc
     5041 cattagttaa aacttgtttt gcagatgatc ttatgaactt tgattttgtt gaagttagac
     5101 actgttttgg acagggagca ttaagagaat acgatagatg tgcaggagaa agagcatacg
     5161 ttattccagc aaaataaact agtagcggcc gctgcaggat atagcaaaag tgggacttaa
     5221 gttcccactt ttgctatttt ataaaaaacg ccctattcgt aatatactat tcaaaactaa
     5281 atattcattg aaatatttgg gtaataatta atattaatct aatatttaga tattgagata
     5341 taataataac atttttttaa aaaatcatct attttcatct taattttata tattaaagcc
     5401 cattttttga gtattcaaat tcaaattatt gtgttattaa catcttatat ataaactttt
     5461 ctatttaatg ttaatgaaaa agtgaatata tatacataga gtaatgttat gatgtatata
     5521 tcaaaaaaaa ctagaggaga cgaccttcca tgacagaata taaaccaaca gttagattag
     5581 caacaagaga tgacgtacct agggcagtaa gaactcttgc agcagctttt gcagattatc
     5641 cagcaacaag acacacagtt gatccagata gacacattga aagagttaca gaattacaag
     5701 aattattttt aacaagagtt ggtcttgata taggtaaagt atgggttgca gatgatggag
     5761 cagcagttgc agtttggaca acaccagaat cagttgaagc tggtgctgtt tttgcagaaa
     5821 ttggaccaag aatggctgaa ttatcaggat caagattagc agctcagcaa cagatggaag
     5881 gattacttgc accacacaga ccaaaagaac cagcatggtt tttagcaaca gttggagttt
     5941 caccagatca ccagggaaaa ggtttaggtt ctgctgttgt acttcctggt gttgaagcag
     6001 ctgaaagagc aggagttcca gcatttttag aaacatcagc accaagaaat cttccatttt
     6061 atgaaagact tggatttaca gttacagcag atgttgaagt tccagaagga ccaagaacat
     6121 ggtgtatgac aagaaaacca ggagcataac catggctccg accgaagcca cccggggcgg
     6181 ccccgccgac cccgcacccg cccccgaggc ccaccgcggg ggacacaccg aacacgccga
     6241 ccctgctgaa cacgcggcgc agttcggtgc ccaggagcgg atcgggaatt aattcgaagc
     6301 tgctggtgaa agagacccta tcttacctgc taaaatctaa gttaattact aatttattat
     6361 taatttatta ttagattggg taaaatagta aaagaaaact aaaggaaacc taatatggtt
     6421 tccttttttt atatattttt aattcactgg ggggatttaa aacttcattt ttaatttaaa
     6481 aggatctagg tgaagatcct tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6541 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     6601 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     6661 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     6721 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     6781 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6841 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6901 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6961 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7021 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7081 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7141 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7201 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7261 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7321 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7381 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7441 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7501 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7561 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7621 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7681 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7741 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7801 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8041 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8521 cctttttacg gttcctggcc ttttgctggc cttttgctca catggtaccc attatttagt
     8581 ttttttttat caccacaacg tataaacccc ctttcagaag aaaacagaga atgaaaacag
     8641 attttaattt atatatcgta tctataattt agtttttttt tattgcagtt ttagaagtgc
     8701 gtttgtcctg gagcgaaata cacgatatcg tgtagtctcc ttcagatgaa aataatattt
     8761 ttaattttca gaaatattac tggaaaaata cctgaaaaat atttgaaaaa aaaacataca
     8821 cccagcgggg aaaaatcgaa cattttgaaa aagtctattt atagggttta aacagggcca
     8881 agacctgcgg gatatacccc aaaaatgttc gataaaaaaa tcgaacattt ttcgaacatt
     8941 ttttataatg ttcgatgtgt gcgagtatgt tcgataaaaa aatcgaacat tttgaaagac
     9001 aatttgaaga gatataaaaa aaataatcta ttataatata gatatataat atgttcgata
     9061 tgttcgatat ttttttatat attatattat atttaacctt gtattgatta ttgaataata
     9121 tttattatta ctttttttga tttcttgtaa aaaatcgaac atattttaca tattgtaaaa
     9181 aataaaaaag atttaaaaaa agtgtccgaa aaagttaaac aaacattatt attaaaatgt
     9241 tagtttaata tgtatcttga tatgctcttc tttctttcat gcttaaactt gtataactgt
     9301 atgctttttt catttccttt attttgtcct ttcctttctt tgttatctgg taatatcctt
     9361 ccttctttcc tgcatacttc aaaaatccaa ggttttttaa gttatcgaga taagaattac
     9421 tagatgaatt cgcgcccggt tctagattta ttcataaaat a
//
