LOCUS       sequence-379176-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged MCRaeoBGA in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  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 8349)
  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-MCRaeo-NG mcrBGA
FEATURES             Location/Qualifiers
     source          1..8349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1770..1793
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1803..1826,1827..1862,1863..1886)
                     /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(5743..5762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5862..5884
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5922..5940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6008..6112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6113..6181,6182..6973)
                     /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(6331..6350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7144..7732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7633..7652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattattac tttttttgat ttcttgtaaa aaatcgaaca tattttacat attgtaaaaa
       61 ataaaaaaga tttaaaaaaa gtgtccgaaa aagttaaaca aacattatta ttaaaatgtt
      121 agtttaatat gtatcttgat atgctcttct ttctttcatg cttaaacttg tataactgta
      181 tgcttttttc atttccttta ttttgtcctt tcctttcttt gttatctggt aatatccttc
      241 cttctttcct gcatacttca aaaatccaag gttttttaag ttatcgagat aagaattact
      301 agatgaattc gcgcccggtt ctagatttat tcataaaata ggaactaaaa taccaaaata
      361 gaaacattta tatacatttg atgtgtaggg tataccataa acctgggagg tgtctcatat
      421 ggttaaatac gaagataaaa ttaaccttta cgatgcaaaa ggaaaccttg cagaagaaaa
      481 cgttccactt gaagcaattt caccattata caatccagtt gttaaagata tggttaaatc
      541 aattaaaaga acagttgcag ttaaccttgc aggaattgaa gcaacactta aagcaggaac
      601 tattggagga aaagcatgta gagttccagg aagaactctt gatttaccaa ttgttgatga
      661 agcagaagca attatggata aagttgaaaa aattcttaaa atttcagaag atgatgatac
      721 aaatattaga actattaatg gaggaaaaca agcagttgtt caggttccat caaaaagact
      781 tgatattgca gcagaatact cagtttcaat gttatcaaca ggaatggcac ttaaagaagc
      841 aattattgat acatttgatg ttgatatgtt tgatggatca actgttcacg cagcaattgt
      901 tggaagatat ccacaagtta tggattacat gggaggaaat attgcatcat tattaggagc
      961 accatcaaat atggaaggat taggatacgc acttagaaac attatggtta accactacgt
     1021 tgcaacaaca aagaaaaatc ttatgaacgc agttgcattt tcatcaatta tggaacaaac
     1081 tgcaatgttt gaaatgggag atgcaattgg atcatttgaa agaatgcact tattaggact
     1141 tggataccag ggacttaacg cagataacat tgttattgat ttagttaaag caaatgcatc
     1201 aggaacagtt ggaactgttg ttgcatcagt tgttgaaaga gcacttgaag ataaagttat
     1261 tgcagaagat aaaacaatga actcaggatt tactatgtac aaaccagttg atgttgcaaa
     1321 atggaatgca tacgcagcag caggattagt tgcagcatca attgttaatt gtggagcagc
     1381 aagagcagca caaaatgttg catcaacaat tctttactac aacgatattc ttgaatatga
     1441 aacaggatta ccaggaactg attacggaag atcagaagga actgcagttg gattttcatt
     1501 tttctcacac tcaatttatg gaggcggagg accaggaatt tttacaggaa atcacgttgt
     1561 tactagacac tcaaaaggat ttgcaattcc accagttaca gcagcattat gtgttgatgc
     1621 aggaacacag atgttttcac cagaaaaaac ttcagcatta gttggagcag tttactcaga
     1681 tattccagaa tttagagaac cacttaaaca cgttattgaa ggagcaattg aaattaaaga
     1741 taaaatttaa attaaaaggt gattctatgg attacaaaga tgatgatgat aaaggcgcca
     1801 gctggtcaca cccacaattt gaaaaaggag gtggatcagg aggtggatca ggaggttcag
     1861 catggtcaca tcctcagttt gaaaaatcag caatggcata tactccacaa ttttgtccag
     1921 gatcaacaac tgttgcacag aatagaagag atcacatgaa tccagatgtt caacttgaaa
     1981 aacttagaga aattccagat gatgaaattg ttaaagttat gggacacaga cagccaggag
     2041 aagattacaa aacagttcac ccaccattag aagaaatgga tatgccagaa gattttgtta
     2101 gagatattgt tgaaccaatg tcaggagcaa aagaaggaca cagaattaga tacattcaat
     2161 ttgcagattc aatgtacaat gcaccagcac agccatatga tagagcaaga acttacatgt
     2221 ggagatttag aggagttgat acaggaactc tttcaggaag acaagttatt gaaatgagag
     2281 aatcaaacct tgaagaaatg tcaaaaaact tttgtattga tacagcattt ttcgatccag
     2341 caacttgtgg aatgagagga gcaacagttc acggacactc acttagactt gatgaaaacg
     2401 gacttatgtt tgatgcatta caaagatatg tttttgatga aaaaacagga cacgttcttt
     2461 acgttaaaga tcaggttgga agaccattag atgaaccagt tgattttgga gaaccattat
     2521 caactgaaaa acttaaagaa attacaacta tttacagaaa agattcaatt gcaatgagag
     2581 atgatgaaga aattgttaca gttgttaaaa gaattcacag agcaagaact cttggaggat
     2641 atatgccaac agaaaaaatt tttgaaggat tataagtgaa atacgataca gcaaatattg
     2701 aatatattac ttaacctacc tctacttacc ttttgaggtg aattttatgg aagcagaaaa
     2761 gaaattattt cttaaagcat taaaagaaaa atttgatgaa gatccaaaag aaaaatacac
     2821 aaaattttac acttacggag gatggaaaca atcaaaaaga aaaactgaat ttgttgaagc
     2881 agcaaaagat attgcagata aaagaggtgg aatgccagga tacaatccag atattggagt
     2941 tccattagga cagagaaaaa tgatgtcata caaactttca ggaacagatg attacgttga
     3001 aggagatgat ttacacttta tgaacaacgc agcaattcaa cagcttcacg atgatattag
     3061 aagaactgtt attgttggaa tggatacagg acacatggtt cttgaaaaac gattaggagt
     3121 tgaagttaca ccagaaacta ttaacgaata catggaaact attaatcacg cattacctgg
     3181 aggagcagtt gttcaagaac acatggttga agttgatcca gcattagcat gggattcata
     3241 cgcaaaaatt tttacaggag atgatgaact tgcagatgaa ttagataaaa gagttttagt
     3301 tgatattaac aaattatttc cagaagaaca agcagaagaa cttaaagcag caattggaaa
     3361 gaaaacatac cagatttcaa gagttccaac attagttgga agagtttgtg atggaggaac
     3421 tattgcaaga tggtcagcaa tgcagattgg aatgtcattt attacagcat acaaactttg
     3481 tgcaggagaa gcagcaattg cagatttttc atacgcagca aaacacgcag atgttattgg
     3541 aatgggaact gcattaccag caagaagatc aagaggagca aatgaacctg gaggaattcc
     3601 atttggaatt ctttgtgatg ttgttcaaac aactagaatt tcagatgatc cagttgaaca
     3661 gtcattagaa gttgttgcaa caggagcaat gttatatgat caaatttggt taggagcata
     3721 catgtctgga ggagttggat ttacacagta tgcaactgca gcatacacag atgatattct
     3781 tgatgatttt tgttactacg gacttgatta cgttgaaaag aaatttggaa gatgtggaac
     3841 aaaagcaact atggatgtag ttgaagatat tgcaacagaa gttactcttt actcacttga
     3901 acaatacgat gaatacccaa ctttattaga agatcacttt ggaggatcac agagagcagc
     3961 agttgcagca gcagcagcag gagtttcagt ttgtatggca acaggaaatt caaatgcagg
     4021 agttaacgga tggtaccttt cacaaattat gcacaaagaa taccactcaa gacttggatt
     4081 ttacggatac gatttacaag atcagtgtgg agcatcaaac tcactttcaa ttagaaacga
     4141 tgaatcatca ccattagaac ttagaggacc aaactaccca aattacgcaa tgaatgttgg
     4201 acaccaagga gaatacgcag gaattactca ggcagcacac tcagcaagag gagatgcatt
     4261 tgcaatgaat ccattaatta aagttgcatt tgcagatcca tcacttattt ttgattttgc
     4321 aagaccaaga aaatgttgtg caagaggagc attaagagaa tttgaagttg caggagaaag
     4381 agatccaatt ttaccagcaa aataaactag tagcggccgc tgcaggatat agcaaaagtg
     4441 ggacttaagt tcccactttt gctattttat aaaaaacgcc ctattcgtaa tatactattc
     4501 aaaactaaat attcattgaa atatttgggt aataattaat attaatctaa tatttagata
     4561 ttgagatata ataataacat ttttttaaaa aatcatctat tttcatctta attttatata
     4621 ttaaagccca ttttttgagt attcaaattc aaattattgt gttattaaca tcttatatat
     4681 aaacttttct atttaatgtt aatgaaaaag tgaatatata tacatagagt aatgttatga
     4741 tgtatatatc aaaaaaaact agaggagacg accttccatg acagaatata aaccaacagt
     4801 tagattagca acaagagatg acgtacctag ggcagtaaga actcttgcag cagcttttgc
     4861 agattatcca gcaacaagac acacagttga tccagataga cacattgaaa gagttacaga
     4921 attacaagaa ttatttttaa caagagttgg tcttgatata ggtaaagtat gggttgcaga
     4981 tgatggagca gcagttgcag tttggacaac accagaatca gttgaagctg gtgctgtttt
     5041 tgcagaaatt ggaccaagaa tggctgaatt atcaggatca agattagcag ctcagcaaca
     5101 gatggaagga ttacttgcac cacacagacc aaaagaacca gcatggtttt tagcaacagt
     5161 tggagtttca ccagatcacc agggaaaagg tttaggttct gctgttgtac ttcctggtgt
     5221 tgaagcagct gaaagagcag gagttccagc atttttagaa acatcagcac caagaaatct
     5281 tccattttat gaaagacttg gatttacagt tacagcagat gttgaagttc cagaaggacc
     5341 aagaacatgg tgtatgacaa gaaaaccagg agcataacca tggctccgac cgaagccacc
     5401 cggggcggcc ccgccgaccc cgcacccgcc cccgaggccc accgcggggg acacaccgaa
     5461 cacgccgacc ctgctgaaca cgcggcgcag ttcggtgccc aggagcggat cgggaattaa
     5521 ttcgaagctg ctggtgaaag agaccctatc ttacctgcta aaatctaagt taattactaa
     5581 tttattatta atttattatt agattgggta aaatagtaaa agaaaactaa aggaaaccta
     5641 atatggtttc ctttttttat atatttttaa ttcactgggg ggatttaaaa cttcattttt
     5701 aatttaaaag gatctaggtg aagatccttg gtgcactctc agtacaatct gctctgatgc
     5761 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5821 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5881 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5941 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6121 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6181 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6241 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6301 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6361 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6421 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6481 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6541 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6601 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6661 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6721 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6781 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6841 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6901 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6961 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7021 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7081 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7141 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7201 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7501 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7561 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7621 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7681 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7741 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tggtacccat
     7801 tatttagttt ttttttatca ccacaacgta taaaccccct ttcagaagaa aacagagaat
     7861 gaaaacagat tttaatttat atatcgtatc tataatttag ttttttttta ttgcagtttt
     7921 agaagtgcgt ttgtcctgga gcgaaataca cgatatcgtg tagtctcctt cagatgaaaa
     7981 taatattttt aattttcaga aatattactg gaaaaatacc tgaaaaatat ttgaaaaaaa
     8041 aacatacacc cagcggggaa aaatcgaaca ttttgaaaaa gtctatttat agggtttaaa
     8101 cagggccaag acctgcggga tataccccaa aaatgttcga taaaaaaatc gaacattttt
     8161 cgaacatttt ttataatgtt cgatgtgtgc gagtatgttc gataaaaaaa tcgaacattt
     8221 tgaaagacaa tttgaagaga tataaaaaaa ataatctatt ataatataga tatataatat
     8281 gttcgatatg ttcgatattt ttttatatat tatattatat ttaaccttgt attgattatt
     8341 gaataatat
//
