LOCUS       sequence-379175-        8961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged MCRaeoBCGA in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8961)
  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 8961)
  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 mcrBCGA
FEATURES             Location/Qualifiers
     source          1..8961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2399..2422
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2432..2455,2456..2491,2492..2515)
                     /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(6372..6391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6491..6513
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6551..6569)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6637..6741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6742..6810,6811..7602)
                     /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(6960..6979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7773..8361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8262..8281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgattattga ataatattta ttattacttt ttttgatttc ttgtaaaaaa tcgaacatat
       61 tttacatatt gtaaaaaata aaaaagattt aaaaaaagtg tccgaaaaag ttaaacaaac
      121 attattatta aaatgttagt ttaatatgta tcttgatatg ctcttctttc tttcatgctt
      181 aaacttgtat aactgtatgc ttttttcatt tcctttattt tgtcctttcc tttctttgtt
      241 atctggtaat atccttcctt ctttcctgca tacttcaaaa atccaaggtt ttttaagtta
      301 tcgagataag aattactaga tgaattcgcg cccggttcta gatttattca taaaatagga
      361 actaaaatac caaaatagaa acatttatat acatttgatg tgtagggtat accataaacc
      421 tgggaggtgt ctcatatggt taaatacgaa gataaaatta acctttacga tgcaaaagga
      481 aaccttgcag aagaaaacgt tccacttgaa gcaatttcac cattatacaa tccagttgtt
      541 aaagatatgg ttaaatcaat taaaagaaca gttgcagtta accttgcagg aattgaagca
      601 acacttaaag caggaactat tggaggaaaa gcatgtagag ttccaggaag aactcttgat
      661 ttaccaattg ttgatgaagc agaagcaatt atggataaag ttgaaaaaat tcttaaaatt
      721 tcagaagatg atgatacaaa tattagaact attaatggag gaaaacaagc agttgttcag
      781 gttccatcaa aaagacttga tattgcagca gaatactcag tttcaatgtt atcaacagga
      841 atggcactta aagaagcaat tattgataca tttgatgttg atatgtttga tggatcaact
      901 gttcacgcag caattgttgg aagatatcca caagttatgg attacatggg aggaaatatt
      961 gcatcattat taggagcacc atcaaatatg gaaggattag gatacgcact tagaaacatt
     1021 atggttaacc actacgttgc aacaacaaag aaaaatctta tgaacgcagt tgcattttca
     1081 tcaattatgg aacaaactgc aatgtttgaa atgggagatg caattggatc atttgaaaga
     1141 atgcacttat taggacttgg ataccaggga cttaacgcag ataacattgt tattgattta
     1201 gttaaagcaa atgcatcagg aacagttgga actgttgttg catcagttgt tgaaagagca
     1261 cttgaagata aagttattgc agaagataaa acaatgaact caggatttac tatgtacaaa
     1321 ccagttgatg ttgcaaaatg gaatgcatac gcagcagcag gattagttgc agcatcaatt
     1381 gttaattgtg gagcagcaag agcagcacaa aatgttgcat caacaattct ttactacaac
     1441 gatattcttg aatatgaaac aggattacca ggaactgatt acggaagatc agaaggaact
     1501 gcagttggat tttcattttt ctcacactca atttatggag gcggaggacc aggaattttt
     1561 acaggaaatc acgttgttac tagacactca aaaggatttg caattccacc agttacagca
     1621 gcattatgtg ttgatgcagg aacacagatg ttttcaccag aaaaaacttc agcattagtt
     1681 ggagcagttt actcagatat tccagaattt agagaaccac ttaaacacgt tattgaagga
     1741 gcaattgaaa ttaaagataa aatttaagag gatagtgatt ttatgccagt tggaagaaaa
     1801 gaacaaattg ttgattgtag agcagttaga ggattaggag aaggtggagg attagcacag
     1861 agaggaacat ttgcagaagg acttaaaaac gatgttgttg ttgttgcaat gtcaccagga
     1921 agaagacaca ttacaaaacc agtttgtgaa attacttacg gaattagaga agcaggaatt
     1981 caaacttcag ttcttgttct taacgcagga tcaggattac cacacgatac accaaaagga
     2041 tcattaggat caacttttgg attaaaacca gaagaagcaa aacaggttaa cagacacaaa
     2101 ctttgtattg ttcaccttgg aaacgttaaa tcacacatta tttacaaagc aagattattt
     2161 cttagatacg ttaaaattcc aacaattatt gtttgtcaaa ctccaattga tatggaagat
     2221 tttgcaaaaa ttggagttaa aacaaaaaac gttatgccag ttaatccaga tacagaagga
     2281 actattgttg atattattac tggagttgtt agaggagaat cagcaacaca gcttaaaatt
     2341 gatgaaacta ttgaaaaaat taaaaatcac ttaaattaaa ttaaaaggtg attctatgga
     2401 ttacaaagat gatgatgata aaggcgccag ctggtcacac ccacaatttg aaaaaggagg
     2461 tggatcagga ggtggatcag gaggttcagc atggtcacat cctcagtttg aaaaatcagc
     2521 aatggcatat actccacaat tttgtccagg atcaacaact gttgcacaga atagaagaga
     2581 tcacatgaat ccagatgttc aacttgaaaa acttagagaa attccagatg atgaaattgt
     2641 taaagttatg ggacacagac agccaggaga agattacaaa acagttcacc caccattaga
     2701 agaaatggat atgccagaag attttgttag agatattgtt gaaccaatgt caggagcaaa
     2761 agaaggacac agaattagat acattcaatt tgcagattca atgtacaatg caccagcaca
     2821 gccatatgat agagcaagaa cttacatgtg gagatttaga ggagttgata caggaactct
     2881 ttcaggaaga caagttattg aaatgagaga atcaaacctt gaagaaatgt caaaaaactt
     2941 ttgtattgat acagcatttt tcgatccagc aacttgtgga atgagaggag caacagttca
     3001 cggacactca cttagacttg atgaaaacgg acttatgttt gatgcattac aaagatatgt
     3061 ttttgatgaa aaaacaggac acgttcttta cgttaaagat caggttggaa gaccattaga
     3121 tgaaccagtt gattttggag aaccattatc aactgaaaaa cttaaagaaa ttacaactat
     3181 ttacagaaaa gattcaattg caatgagaga tgatgaagaa attgttacag ttgttaaaag
     3241 aattcacaga gcaagaactc ttggaggata tatgccaaca gaaaaaattt ttgaaggatt
     3301 ataagtgaaa tacgatacag caaatattga atatattact taacctacct ctacttacct
     3361 tttgaggtga attttatgga agcagaaaag aaattatttc ttaaagcatt aaaagaaaaa
     3421 tttgatgaag atccaaaaga aaaatacaca aaattttaca cttacggagg atggaaacaa
     3481 tcaaaaagaa aaactgaatt tgttgaagca gcaaaagata ttgcagataa aagaggtgga
     3541 atgccaggat acaatccaga tattggagtt ccattaggac agagaaaaat gatgtcatac
     3601 aaactttcag gaacagatga ttacgttgaa ggagatgatt tacactttat gaacaacgca
     3661 gcaattcaac agcttcacga tgatattaga agaactgtta ttgttggaat ggatacagga
     3721 cacatggttc ttgaaaaacg attaggagtt gaagttacac cagaaactat taacgaatac
     3781 atggaaacta ttaatcacgc attacctgga ggagcagttg ttcaagaaca catggttgaa
     3841 gttgatccag cattagcatg ggattcatac gcaaaaattt ttacaggaga tgatgaactt
     3901 gcagatgaat tagataaaag agttttagtt gatattaaca aattatttcc agaagaacaa
     3961 gcagaagaac ttaaagcagc aattggaaag aaaacatacc agatttcaag agttccaaca
     4021 ttagttggaa gagtttgtga tggaggaact attgcaagat ggtcagcaat gcagattgga
     4081 atgtcattta ttacagcata caaactttgt gcaggagaag cagcaattgc agatttttca
     4141 tacgcagcaa aacacgcaga tgttattgga atgggaactg cattaccagc aagaagatca
     4201 agaggagcaa atgaacctgg aggaattcca tttggaattc tttgtgatgt tgttcaaaca
     4261 actagaattt cagatgatcc agttgaacag tcattagaag ttgttgcaac aggagcaatg
     4321 ttatatgatc aaatttggtt aggagcatac atgtctggag gagttggatt tacacagtat
     4381 gcaactgcag catacacaga tgatattctt gatgattttt gttactacgg acttgattac
     4441 gttgaaaaga aatttggaag atgtggaaca aaagcaacta tggatgtagt tgaagatatt
     4501 gcaacagaag ttactcttta ctcacttgaa caatacgatg aatacccaac tttattagaa
     4561 gatcactttg gaggatcaca gagagcagca gttgcagcag cagcagcagg agtttcagtt
     4621 tgtatggcaa caggaaattc aaatgcagga gttaacggat ggtacctttc acaaattatg
     4681 cacaaagaat accactcaag acttggattt tacggatacg atttacaaga tcagtgtgga
     4741 gcatcaaact cactttcaat tagaaacgat gaatcatcac cattagaact tagaggacca
     4801 aactacccaa attacgcaat gaatgttgga caccaaggag aatacgcagg aattactcag
     4861 gcagcacact cagcaagagg agatgcattt gcaatgaatc cattaattaa agttgcattt
     4921 gcagatccat cacttatttt tgattttgca agaccaagaa aatgttgtgc aagaggagca
     4981 ttaagagaat ttgaagttgc aggagaaaga gatccaattt taccagcaaa ataaactagt
     5041 agcggccgct gcaggatata gcaaaagtgg gacttaagtt cccacttttg ctattttata
     5101 aaaaacgccc tattcgtaat atactattca aaactaaata ttcattgaaa tatttgggta
     5161 ataattaata ttaatctaat atttagatat tgagatataa taataacatt tttttaaaaa
     5221 atcatctatt ttcatcttaa ttttatatat taaagcccat tttttgagta ttcaaattca
     5281 aattattgtg ttattaacat cttatatata aacttttcta tttaatgtta atgaaaaagt
     5341 gaatatatat acatagagta atgttatgat gtatatatca aaaaaaacta gaggagacga
     5401 ccttccatga cagaatataa accaacagtt agattagcaa caagagatga cgtacctagg
     5461 gcagtaagaa ctcttgcagc agcttttgca gattatccag caacaagaca cacagttgat
     5521 ccagatagac acattgaaag agttacagaa ttacaagaat tatttttaac aagagttggt
     5581 cttgatatag gtaaagtatg ggttgcagat gatggagcag cagttgcagt ttggacaaca
     5641 ccagaatcag ttgaagctgg tgctgttttt gcagaaattg gaccaagaat ggctgaatta
     5701 tcaggatcaa gattagcagc tcagcaacag atggaaggat tacttgcacc acacagacca
     5761 aaagaaccag catggttttt agcaacagtt ggagtttcac cagatcacca gggaaaaggt
     5821 ttaggttctg ctgttgtact tcctggtgtt gaagcagctg aaagagcagg agttccagca
     5881 tttttagaaa catcagcacc aagaaatctt ccattttatg aaagacttgg atttacagtt
     5941 acagcagatg ttgaagttcc agaaggacca agaacatggt gtatgacaag aaaaccagga
     6001 gcataaccat ggctccgacc gaagccaccc ggggcggccc cgccgacccc gcacccgccc
     6061 ccgaggccca ccgcggggga cacaccgaac acgccgaccc tgctgaacac gcggcgcagt
     6121 tcggtgccca ggagcggatc gggaattaat tcgaagctgc tggtgaaaga gaccctatct
     6181 tacctgctaa aatctaagtt aattactaat ttattattaa tttattatta gattgggtaa
     6241 aatagtaaaa gaaaactaaa ggaaacctaa tatggtttcc tttttttata tatttttaat
     6301 tcactggggg gatttaaaac ttcattttta atttaaaagg atctaggtga agatccttgg
     6361 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6421 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6481 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6541 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6601 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6721 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6781 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6841 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6901 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6961 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     7021 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7081 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7141 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7201 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7261 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7321 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7381 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7441 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7501 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7561 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7621 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7681 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7741 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7801 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7861 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7921 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7981 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8041 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8101 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8161 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8221 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8281 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8341 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8401 tgctggcctt ttgctcacat ggtacccatt atttagtttt tttttatcac cacaacgtat
     8461 aaaccccctt tcagaagaaa acagagaatg aaaacagatt ttaatttata tatcgtatct
     8521 ataatttagt ttttttttat tgcagtttta gaagtgcgtt tgtcctggag cgaaatacac
     8581 gatatcgtgt agtctccttc agatgaaaat aatattttta attttcagaa atattactgg
     8641 aaaaatacct gaaaaatatt tgaaaaaaaa acatacaccc agcggggaaa aatcgaacat
     8701 tttgaaaaag tctatttata gggtttaaac agggccaaga cctgcgggat ataccccaaa
     8761 aatgttcgat aaaaaaatcg aacatttttc gaacattttt tataatgttc gatgtgtgcg
     8821 agtatgttcg ataaaaaaat cgaacatttt gaaagacaat ttgaagagat ataaaaaaaa
     8881 taatctatta taatatagat atataatatg ttcgatatgt tcgatatttt tttatatatt
     8941 atattatatt taaccttgta t
//
