LOCUS       sequence-379178-        9512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ND-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-ND
FEATURES             Location/Qualifiers
     source          1..9512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2208..2231
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2241..2264,2265..2300,2301..2324)
                     /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(7301..7320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7420..7442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7480..7498)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7566..7670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7671..7739,7740..8531)
                     /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(7889..7908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8702..9290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9191..9210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcgtgtagt ctccttcaga tgaaaataat atttttaatt ttcagaaata ttactggaaa
       61 aatacctgaa aaatatttga aaaaaaaaca tacacccagc ggggaaaaat cgaacatttt
      121 gaaaaagtct atttataggg tttaaacagg gccaagacct gcgggatata ccccaaaaat
      181 gttcgataaa aaaatcgaac atttttcgaa cattttttat aatgttcgat gtgtgcgagt
      241 atgttcgata aaaaaatcga acattttgaa agacaatttg aagagatata aaaaaaataa
      301 tctattataa tatagatata taatatgttc gatatgttcg atattttttt atatattata
      361 ttatatttaa ccttgtattg attattgaat aatatttatt attacttttt ttgatttctt
      421 gtaaaaaatc gaacatattt tacatattgt aaaaaataaa aaagatttaa aaaaagtgtc
      481 cgaaaaagtt aaacaaacat tattattaaa atgttagttt aatatgtatc ttgatatgct
      541 cttctttctt tcatgcttaa acttgtataa ctgtatgctt ttttcatttc ctttattttg
      601 tcctttcctt tctttgttat ctggtaatat ccttccttct ttcctgcata cttcaaaaat
      661 ccaaggtttt ttaagttatc gagataagaa ttactagatg aattcgcgcc cggttctaga
      721 tttattcata aaataggaac taaaatacca aaatagaaac atttatatac atttgatgtg
      781 tagggtatac cataaacctg ggaggtgtct catatggtta aatacgaaga taaaattaac
      841 ctttacgatg caaaaggaaa ccttgcagaa gaaaacgttc cacttgaagc aatttcacca
      901 ttatacaatc cagttgttaa agatatggtt aaatcaatta aaagaacagt tgcagttaac
      961 cttgcaggaa ttgaagcaac acttaaagca ggaactattg gaggaaaagc atgtagagtt
     1021 ccaggaagaa ctcttgattt accaattgtt gatgaagcag aagcaattat ggataaagtt
     1081 gaaaaaattc ttaaaatttc agaagatgat gatacaaata ttagaactat taatggagga
     1141 aaacaagcag ttgttcaggt tccatcaaaa agacttgata ttgcagcaga atactcagtt
     1201 tcaatgttat caacaggaat ggcacttaaa gaagcaatta ttgatacatt tgatgttgat
     1261 atgtttgatg gatcaactgt tcacgcagca attgttggaa gatatccaca agttatggat
     1321 tacatgggag gaaatattgc atcattatta ggagcaccat caaatatgga aggattagga
     1381 tacgcactta gaaacattat ggttaaccac tacgttgcaa caacaaagaa aaatcttatg
     1441 aacgcagttg cattttcatc aattatggaa caaactgcaa tgtttgaaat gggagatgca
     1501 attggatcat ttgaaagaat gcacttatta ggacttggat accagggact taacgcagat
     1561 aacattgtta ttgatttagt taaagcaaat gcatcaggaa cagttggaac tgttgttgca
     1621 tcagttgttg aaagagcact tgaagataaa gttattgcag aagataaaac aatgaactca
     1681 ggatttacta tgtacaaacc agttgatgtt gcaaaatgga atgcatacgc agcagcagga
     1741 ttagttgcag catcaattgt taattgtgga gcagcaagag cagcacaaaa tgttgcatca
     1801 acaattcttt actacaacga tattcttgaa tatgaaacag gattaccagg aactgattac
     1861 ggaagatcag aaggaactgc agttggattt tcatttttct cacactcaat ttatggaggc
     1921 ggaggaccag gaatttttac aggaaatcac gttgttacta gacactcaaa aggatttgca
     1981 attccaccag ttacagcagc attatgtgtt gatgcaggaa cacagatgtt ttcaccagaa
     2041 aaaacttcag cattagttgg agcagtttac tcagatattc cagaatttag agaaccactt
     2101 aaacacgtta ttgaaggagc aattgaaatt aaagataaaa tttaaattat caattgctca
     2161 tatatctttt attcccggta tttaacttat tggtaggtaa tattatggat tacaaagatg
     2221 atgatgataa aggcgccagc tggtcacacc cacaatttga aaaaggaggt ggatcaggag
     2281 gtggatcagg aggttcagca tggtcacatc ctcagtttga aaaatcagca atgattgaaa
     2341 ttgaagtttt tccacacaga tttttaaaag cagatacaac tgaaaaattt cttaacgata
     2401 tttactcaat tgaaactgtt gaaagagtta ttgttcacgg acaaccatta ccaaaaactg
     2461 tttattacgg accagcaaga ggaacaccag ttaatcaccc agaaagaaaa gaagttcttg
     2521 ttaacggagt tccagttgaa cttacagtta tggcaggaag attttggatt cttcttgaaa
     2581 acgataacga acttaacaaa attgaagaaa tttgtaaatc attatttcca tatggataca
     2641 atttaaatgt tggaaaattt actagagatt caccaacaac tacagattac atgaaatacg
     2701 gagaaaaact tgttaactca atggatagaa aaatgattgg acttacagat ccaagatcaa
     2761 aatttgcatc atcagttaaa atgattccaa aaaacgaaaa cgcagaatgt gaacagaatt
     2821 aagaggatag tgattttatg ccagttggaa gaaaagaaca aattgttgat tgtagagcag
     2881 ttagaggatt aggagaaggt ggaggattag cacagagagg aacatttgca gaaggactta
     2941 aaaacgatgt tgttgttgtt gcaatgtcac caggaagaag acacattaca aaaccagttt
     3001 gtgaaattac ttacggaatt agagaagcag gaattcaaac ttcagttctt gttcttaacg
     3061 caggatcagg attaccacac gatacaccaa aaggatcatt aggatcaact tttggattaa
     3121 aaccagaaga agcaaaacag gttaacagac acaaactttg tattgttcac cttggaaacg
     3181 ttaaatcaca cattatttac aaagcaagat tatttcttag atacgttaaa attccaacaa
     3241 ttattgtttg tcaaactcca attgatatgg aagattttgc aaaaattgga gttaaaacaa
     3301 aaaacgttat gccagttaat ccagatacag aaggaactat tgttgatatt attactggag
     3361 ttgttagagg agaatcagca acacagctta aaattgatga aactattgaa aaaattaaaa
     3421 atcacttaaa ttaaattaaa aggtgattct atggcatata ctccacaatt ttgtccagga
     3481 tcaacaactg ttgcacagaa tagaagagat cacatgaatc cagatgttca acttgaaaaa
     3541 cttagagaaa ttccagatga tgaaattgtt aaagttatgg gacacagaca gccaggagaa
     3601 gattacaaaa cagttcaccc accattagaa gaaatggata tgccagaaga ttttgttaga
     3661 gatattgttg aaccaatgtc aggagcaaaa gaaggacaca gaattagata cattcaattt
     3721 gcagattcaa tgtacaatgc accagcacag ccatatgata gagcaagaac ttacatgtgg
     3781 agatttagag gagttgatac aggaactctt tcaggaagac aagttattga aatgagagaa
     3841 tcaaaccttg aagaaatgtc aaaaaacttt tgtattgata cagcattttt cgatccagca
     3901 acttgtggaa tgagaggagc aacagttcac ggacactcac ttagacttga tgaaaacgga
     3961 cttatgtttg atgcattaca aagatatgtt tttgatgaaa aaacaggaca cgttctttac
     4021 gttaaagatc aggttggaag accattagat gaaccagttg attttggaga accattatca
     4081 actgaaaaac ttaaagaaat tacaactatt tacagaaaag attcaattgc aatgagagat
     4141 gatgaagaaa ttgttacagt tgttaaaaga attcacagag caagaactct tggaggatat
     4201 atgccaacag aaaaaatttt tgaaggatta taagtgaaat acgatacagc aaatattgaa
     4261 tatattactt aacctacctc tacttacctt ttgaggtgaa ttttatggaa gcagaaaaga
     4321 aattatttct taaagcatta aaagaaaaat ttgatgaaga tccaaaagaa aaatacacaa
     4381 aattttacac ttacggagga tggaaacaat caaaaagaaa aactgaattt gttgaagcag
     4441 caaaagatat tgcagataaa agaggtggaa tgccaggata caatccagat attggagttc
     4501 cattaggaca gagaaaaatg atgtcataca aactttcagg aacagatgat tacgttgaag
     4561 gagatgattt acactttatg aacaacgcag caattcaaca gcttcacgat gatattagaa
     4621 gaactgttat tgttggaatg gatacaggac acatggttct tgaaaaacga ttaggagttg
     4681 aagttacacc agaaactatt aacgaataca tggaaactat taatcacgca ttacctggag
     4741 gagcagttgt tcaagaacac atggttgaag ttgatccagc attagcatgg gattcatacg
     4801 caaaaatttt tacaggagat gatgaacttg cagatgaatt agataaaaga gttttagttg
     4861 atattaacaa attatttcca gaagaacaag cagaagaact taaagcagca attggaaaga
     4921 aaacatacca gatttcaaga gttccaacat tagttggaag agtttgtgat ggaggaacta
     4981 ttgcaagatg gtcagcaatg cagattggaa tgtcatttat tacagcatac aaactttgtg
     5041 caggagaagc agcaattgca gatttttcat acgcagcaaa acacgcagat gttattggaa
     5101 tgggaactgc attaccagca agaagatcaa gaggagcaaa tgaacctgga ggaattccat
     5161 ttggaattct ttgtgatgtt gttcaaacaa ctagaatttc agatgatcca gttgaacagt
     5221 cattagaagt tgttgcaaca ggagcaatgt tatatgatca aatttggtta ggagcataca
     5281 tgtctggagg agttggattt acacagtatg caactgcagc atacacagat gatattcttg
     5341 atgatttttg ttactacgga cttgattacg ttgaaaagaa atttggaaga tgtggaacaa
     5401 aagcaactat ggatgtagtt gaagatattg caacagaagt tactctttac tcacttgaac
     5461 aatacgatga atacccaact ttattagaag atcactttgg aggatcacag agagcagcag
     5521 ttgcagcagc agcagcagga gtttcagttt gtatggcaac aggaaattca aatgcaggag
     5581 ttaacggatg gtacctttca caaattatgc acaaagaata ccactcaaga cttggatttt
     5641 acggatacga tttacaagat cagtgtggag catcaaactc actttcaatt agaaacgatg
     5701 aatcatcacc attagaactt agaggaccaa actacccaaa ttacgcaatg aatgttggac
     5761 accaaggaga atacgcagga attactcagg cagcacactc agcaagagga gatgcatttg
     5821 caatgaatcc attaattaaa gttgcatttg cagatccatc acttattttt gattttgcaa
     5881 gaccaagaaa atgttgtgca agaggagcat taagagaatt tgaagttgca ggagaaagag
     5941 atccaatttt accagcaaaa taaactagta gcggccgctg caggatatag caaaagtggg
     6001 acttaagttc ccacttttgc tattttataa aaaacgccct attcgtaata tactattcaa
     6061 aactaaatat tcattgaaat atttgggtaa taattaatat taatctaata tttagatatt
     6121 gagatataat aataacattt ttttaaaaaa tcatctattt tcatcttaat tttatatatt
     6181 aaagcccatt ttttgagtat tcaaattcaa attattgtgt tattaacatc ttatatataa
     6241 acttttctat ttaatgttaa tgaaaaagtg aatatatata catagagtaa tgttatgatg
     6301 tatatatcaa aaaaaactag aggagacgac cttccatgac agaatataaa ccaacagtta
     6361 gattagcaac aagagatgac gtacctaggg cagtaagaac tcttgcagca gcttttgcag
     6421 attatccagc aacaagacac acagttgatc cagatagaca cattgaaaga gttacagaat
     6481 tacaagaatt atttttaaca agagttggtc ttgatatagg taaagtatgg gttgcagatg
     6541 atggagcagc agttgcagtt tggacaacac cagaatcagt tgaagctggt gctgtttttg
     6601 cagaaattgg accaagaatg gctgaattat caggatcaag attagcagct cagcaacaga
     6661 tggaaggatt acttgcacca cacagaccaa aagaaccagc atggttttta gcaacagttg
     6721 gagtttcacc agatcaccag ggaaaaggtt taggttctgc tgttgtactt cctggtgttg
     6781 aagcagctga aagagcagga gttccagcat ttttagaaac atcagcacca agaaatcttc
     6841 cattttatga aagacttgga tttacagtta cagcagatgt tgaagttcca gaaggaccaa
     6901 gaacatggtg tatgacaaga aaaccaggag cataaccatg gctccgaccg aagccacccg
     6961 gggcggcccc gccgaccccg cacccgcccc cgaggcccac cgcgggggac acaccgaaca
     7021 cgccgaccct gctgaacacg cggcgcagtt cggtgcccag gagcggatcg ggaattaatt
     7081 cgaagctgct ggtgaaagag accctatctt acctgctaaa atctaagtta attactaatt
     7141 tattattaat ttattattag attgggtaaa atagtaaaag aaaactaaag gaaacctaat
     7201 atggtttcct ttttttatat atttttaatt cactgggggg atttaaaact tcatttttaa
     7261 tttaaaagga tctaggtgaa gatccttggt gcactctcag tacaatctgc tctgatgccg
     7321 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     7381 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     7441 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     7501 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     7561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     8041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     8101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     8161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     8221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     8401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     8881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     9001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     9061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     9121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     9181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     9241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     9301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gtacccatta
     9361 tttagttttt ttttatcacc acaacgtata aacccccttt cagaagaaaa cagagaatga
     9421 aaacagattt taatttatat atcgtatcta taatttagtt tttttttatt gcagttttag
     9481 aagtgcgttt gtcctggagc gaaatacacg at
//
