LOCUS       sequence-379177-        9512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NC-tagged MCRaeo in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9512)
  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 9512)
  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-NC
FEATURES             Location/Qualifiers
     source          1..9512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2136..2159
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2169..2192,2193..2228,2229..2252)
                     /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(6722..6741)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6841..6863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6901..6919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6987..7091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7092..7160,7161..7952)
                     /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(7310..7329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8123..8711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8612..8631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttcattt cctttatttt gtcctttcct ttctttgtta tctggtaata tccttccttc
       61 tttcctgcat acttcaaaaa tccaaggttt tttaagttat cgagataaga attactagat
      121 gaattcgcgc ccggttctag atttattcat aaaataggaa ctaaaatacc aaaatagaaa
      181 catttatata catttgatgt gtagggtata ccataaacct gggaggtgtc tcatatggtt
      241 aaatacgaag ataaaattaa cctttacgat gcaaaaggaa accttgcaga agaaaacgtt
      301 ccacttgaag caatttcacc attatacaat ccagttgtta aagatatggt taaatcaatt
      361 aaaagaacag ttgcagttaa ccttgcagga attgaagcaa cacttaaagc aggaactatt
      421 ggaggaaaag catgtagagt tccaggaaga actcttgatt taccaattgt tgatgaagca
      481 gaagcaatta tggataaagt tgaaaaaatt cttaaaattt cagaagatga tgatacaaat
      541 attagaacta ttaatggagg aaaacaagca gttgttcagg ttccatcaaa aagacttgat
      601 attgcagcag aatactcagt ttcaatgtta tcaacaggaa tggcacttaa agaagcaatt
      661 attgatacat ttgatgttga tatgtttgat ggatcaactg ttcacgcagc aattgttgga
      721 agatatccac aagttatgga ttacatggga ggaaatattg catcattatt aggagcacca
      781 tcaaatatgg aaggattagg atacgcactt agaaacatta tggttaacca ctacgttgca
      841 acaacaaaga aaaatcttat gaacgcagtt gcattttcat caattatgga acaaactgca
      901 atgtttgaaa tgggagatgc aattggatca tttgaaagaa tgcacttatt aggacttgga
      961 taccagggac ttaacgcaga taacattgtt attgatttag ttaaagcaaa tgcatcagga
     1021 acagttggaa ctgttgttgc atcagttgtt gaaagagcac ttgaagataa agttattgca
     1081 gaagataaaa caatgaactc aggatttact atgtacaaac cagttgatgt tgcaaaatgg
     1141 aatgcatacg cagcagcagg attagttgca gcatcaattg ttaattgtgg agcagcaaga
     1201 gcagcacaaa atgttgcatc aacaattctt tactacaacg atattcttga atatgaaaca
     1261 ggattaccag gaactgatta cggaagatca gaaggaactg cagttggatt ttcatttttc
     1321 tcacactcaa tttatggagg cggaggacca ggaattttta caggaaatca cgttgttact
     1381 agacactcaa aaggatttgc aattccacca gttacagcag cattatgtgt tgatgcagga
     1441 acacagatgt tttcaccaga aaaaacttca gcattagttg gagcagttta ctcagatatt
     1501 ccagaattta gagaaccact taaacacgtt attgaaggag caattgaaat taaagataaa
     1561 atttaaatta tcaattgctc atatatcttt tattcccggt atttaactta ttggtaggta
     1621 atattatgat tgaaattgaa gtttttccac acagattttt aaaagcagat acaactgaaa
     1681 aatttcttaa cgatatttac tcaattgaaa ctgttgaaag agttattgtt cacggacaac
     1741 cattaccaaa aactgtttat tacggaccag caagaggaac accagttaat cacccagaaa
     1801 gaaaagaagt tcttgttaac ggagttccag ttgaacttac agttatggca ggaagatttt
     1861 ggattcttct tgaaaacgat aacgaactta acaaaattga agaaatttgt aaatcattat
     1921 ttccatatgg atacaattta aatgttggaa aatttactag agattcacca acaactacag
     1981 attacatgaa atacggagaa aaacttgtta actcaatgga tagaaaaatg attggactta
     2041 cagatccaag atcaaaattt gcatcatcag ttaaaatgat tccaaaaaac gaaaacgcag
     2101 aatgtgaaca gaattaagag gatagtgatt ttatggatta caaagatgat gatgataaag
     2161 gcgccagctg gtcacaccca caatttgaaa aaggaggtgg atcaggaggt ggatcaggag
     2221 gttcagcatg gtcacatcct cagtttgaaa aatcagcaat gccagttgga agaaaagaac
     2281 aaattgttga ttgtagagca gttagaggat taggagaagg tggaggatta gcacagagag
     2341 gaacatttgc agaaggactt aaaaacgatg ttgttgttgt tgcaatgtca ccaggaagaa
     2401 gacacattac aaaaccagtt tgtgaaatta cttacggaat tagagaagca ggaattcaaa
     2461 cttcagttct tgttcttaac gcaggatcag gattaccaca cgatacacca aaaggatcat
     2521 taggatcaac ttttggatta aaaccagaag aagcaaaaca ggttaacaga cacaaacttt
     2581 gtattgttca ccttggaaac gttaaatcac acattattta caaagcaaga ttatttctta
     2641 gatacgttaa aattccaaca attattgttt gtcaaactcc aattgatatg gaagattttg
     2701 caaaaattgg agttaaaaca aaaaacgtta tgccagttaa tccagataca gaaggaacta
     2761 ttgttgatat tattactgga gttgttagag gagaatcagc aacacagctt aaaattgatg
     2821 aaactattga aaaaattaaa aatcacttaa attaaattaa aaggtgattc tatggcatat
     2881 actccacaat tttgtccagg atcaacaact gttgcacaga atagaagaga tcacatgaat
     2941 ccagatgttc aacttgaaaa acttagagaa attccagatg atgaaattgt taaagttatg
     3001 ggacacagac agccaggaga agattacaaa acagttcacc caccattaga agaaatggat
     3061 atgccagaag attttgttag agatattgtt gaaccaatgt caggagcaaa agaaggacac
     3121 agaattagat acattcaatt tgcagattca atgtacaatg caccagcaca gccatatgat
     3181 agagcaagaa cttacatgtg gagatttaga ggagttgata caggaactct ttcaggaaga
     3241 caagttattg aaatgagaga atcaaacctt gaagaaatgt caaaaaactt ttgtattgat
     3301 acagcatttt tcgatccagc aacttgtgga atgagaggag caacagttca cggacactca
     3361 cttagacttg atgaaaacgg acttatgttt gatgcattac aaagatatgt ttttgatgaa
     3421 aaaacaggac acgttcttta cgttaaagat caggttggaa gaccattaga tgaaccagtt
     3481 gattttggag aaccattatc aactgaaaaa cttaaagaaa ttacaactat ttacagaaaa
     3541 gattcaattg caatgagaga tgatgaagaa attgttacag ttgttaaaag aattcacaga
     3601 gcaagaactc ttggaggata tatgccaaca gaaaaaattt ttgaaggatt ataagtgaaa
     3661 tacgatacag caaatattga atatattact taacctacct ctacttacct tttgaggtga
     3721 attttatgga agcagaaaag aaattatttc ttaaagcatt aaaagaaaaa tttgatgaag
     3781 atccaaaaga aaaatacaca aaattttaca cttacggagg atggaaacaa tcaaaaagaa
     3841 aaactgaatt tgttgaagca gcaaaagata ttgcagataa aagaggtgga atgccaggat
     3901 acaatccaga tattggagtt ccattaggac agagaaaaat gatgtcatac aaactttcag
     3961 gaacagatga ttacgttgaa ggagatgatt tacactttat gaacaacgca gcaattcaac
     4021 agcttcacga tgatattaga agaactgtta ttgttggaat ggatacagga cacatggttc
     4081 ttgaaaaacg attaggagtt gaagttacac cagaaactat taacgaatac atggaaacta
     4141 ttaatcacgc attacctgga ggagcagttg ttcaagaaca catggttgaa gttgatccag
     4201 cattagcatg ggattcatac gcaaaaattt ttacaggaga tgatgaactt gcagatgaat
     4261 tagataaaag agttttagtt gatattaaca aattatttcc agaagaacaa gcagaagaac
     4321 ttaaagcagc aattggaaag aaaacatacc agatttcaag agttccaaca ttagttggaa
     4381 gagtttgtga tggaggaact attgcaagat ggtcagcaat gcagattgga atgtcattta
     4441 ttacagcata caaactttgt gcaggagaag cagcaattgc agatttttca tacgcagcaa
     4501 aacacgcaga tgttattgga atgggaactg cattaccagc aagaagatca agaggagcaa
     4561 atgaacctgg aggaattcca tttggaattc tttgtgatgt tgttcaaaca actagaattt
     4621 cagatgatcc agttgaacag tcattagaag ttgttgcaac aggagcaatg ttatatgatc
     4681 aaatttggtt aggagcatac atgtctggag gagttggatt tacacagtat gcaactgcag
     4741 catacacaga tgatattctt gatgattttt gttactacgg acttgattac gttgaaaaga
     4801 aatttggaag atgtggaaca aaagcaacta tggatgtagt tgaagatatt gcaacagaag
     4861 ttactcttta ctcacttgaa caatacgatg aatacccaac tttattagaa gatcactttg
     4921 gaggatcaca gagagcagca gttgcagcag cagcagcagg agtttcagtt tgtatggcaa
     4981 caggaaattc aaatgcagga gttaacggat ggtacctttc acaaattatg cacaaagaat
     5041 accactcaag acttggattt tacggatacg atttacaaga tcagtgtgga gcatcaaact
     5101 cactttcaat tagaaacgat gaatcatcac cattagaact tagaggacca aactacccaa
     5161 attacgcaat gaatgttgga caccaaggag aatacgcagg aattactcag gcagcacact
     5221 cagcaagagg agatgcattt gcaatgaatc cattaattaa agttgcattt gcagatccat
     5281 cacttatttt tgattttgca agaccaagaa aatgttgtgc aagaggagca ttaagagaat
     5341 ttgaagttgc aggagaaaga gatccaattt taccagcaaa ataaactagt agcggccgct
     5401 gcaggatata gcaaaagtgg gacttaagtt cccacttttg ctattttata aaaaacgccc
     5461 tattcgtaat atactattca aaactaaata ttcattgaaa tatttgggta ataattaata
     5521 ttaatctaat atttagatat tgagatataa taataacatt tttttaaaaa atcatctatt
     5581 ttcatcttaa ttttatatat taaagcccat tttttgagta ttcaaattca aattattgtg
     5641 ttattaacat cttatatata aacttttcta tttaatgtta atgaaaaagt gaatatatat
     5701 acatagagta atgttatgat gtatatatca aaaaaaacta gaggagacga ccttccatga
     5761 cagaatataa accaacagtt agattagcaa caagagatga cgtacctagg gcagtaagaa
     5821 ctcttgcagc agcttttgca gattatccag caacaagaca cacagttgat ccagatagac
     5881 acattgaaag agttacagaa ttacaagaat tatttttaac aagagttggt cttgatatag
     5941 gtaaagtatg ggttgcagat gatggagcag cagttgcagt ttggacaaca ccagaatcag
     6001 ttgaagctgg tgctgttttt gcagaaattg gaccaagaat ggctgaatta tcaggatcaa
     6061 gattagcagc tcagcaacag atggaaggat tacttgcacc acacagacca aaagaaccag
     6121 catggttttt agcaacagtt ggagtttcac cagatcacca gggaaaaggt ttaggttctg
     6181 ctgttgtact tcctggtgtt gaagcagctg aaagagcagg agttccagca tttttagaaa
     6241 catcagcacc aagaaatctt ccattttatg aaagacttgg atttacagtt acagcagatg
     6301 ttgaagttcc agaaggacca agaacatggt gtatgacaag aaaaccagga gcataaccat
     6361 ggctccgacc gaagccaccc ggggcggccc cgccgacccc gcacccgccc ccgaggccca
     6421 ccgcggggga cacaccgaac acgccgaccc tgctgaacac gcggcgcagt tcggtgccca
     6481 ggagcggatc gggaattaat tcgaagctgc tggtgaaaga gaccctatct tacctgctaa
     6541 aatctaagtt aattactaat ttattattaa tttattatta gattgggtaa aatagtaaaa
     6601 gaaaactaaa ggaaacctaa tatggtttcc tttttttata tatttttaat tcactggggg
     6661 gatttaaaac ttcattttta atttaaaagg atctaggtga agatccttgg tgcactctca
     6721 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6781 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6841 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6901 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7381 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7441 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7501 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7561 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7621 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7681 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7741 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7801 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7861 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7921 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7981 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8041 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8101 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     8161 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     8221 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     8281 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     8341 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8401 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     8461 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8521 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8581 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8641 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8701 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8761 ttgctcacat ggtacccatt atttagtttt tttttatcac cacaacgtat aaaccccctt
     8821 tcagaagaaa acagagaatg aaaacagatt ttaatttata tatcgtatct ataatttagt
     8881 ttttttttat tgcagtttta gaagtgcgtt tgtcctggag cgaaatacac gatatcgtgt
     8941 agtctccttc agatgaaaat aatattttta attttcagaa atattactgg aaaaatacct
     9001 gaaaaatatt tgaaaaaaaa acatacaccc agcggggaaa aatcgaacat tttgaaaaag
     9061 tctatttata gggtttaaac agggccaaga cctgcgggat ataccccaaa aatgttcgat
     9121 aaaaaaatcg aacatttttc gaacattttt tataatgttc gatgtgtgcg agtatgttcg
     9181 ataaaaaaat cgaacatttt gaaagacaat ttgaagagat ataaaaaaaa taatctatta
     9241 taatatagat atataatatg ttcgatatgt tcgatatttt tttatatatt atattatatt
     9301 taaccttgta ttgattattg aataatattt attattactt tttttgattt cttgtaaaaa
     9361 atcgaacata ttttacatat tgtaaaaaat aaaaaagatt taaaaaaagt gtccgaaaaa
     9421 gttaaacaaa cattattatt aaaatgttag tttaatatgt atcttgatat gctcttcttt
     9481 ctttcatgct taaacttgta taactgtatg ct
//
