LOCUS       sequence-379061-        9416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged MCRmar in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9416)
  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 9416)
  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-MCRmar-NG
FEATURES             Location/Qualifiers
     source          1..9416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2868..2891
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2901..2924,2925..2960,2961..2984)
                     /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(6820..6839)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6939..6961
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6999..7017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7085..7189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7190..7258,7259..8050)
                     /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(7408..7427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8221..8809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8710..8729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgaataatat ttattattac tttttttgat ttcttgtaaa aaatcgaaca tattttacat
       61 attgtaaaaa ataaaaaaga tttaaaaaaa gtgtccgaaa aagttaaaca aacattatta
      121 ttaaaatgtt agtttaatat gtatcttgat atgctcttct ttctttcatg cttaaacttg
      181 tataactgta tgcttttttc atttccttta ttttgtcctt tcctttcttt gttatctggt
      241 aatatccttc cttctttcct gcatacttca aaaatccaag gttttttaag ttacgagata
      301 agaattacta gatgaattcg cgcccggttc tagatttatt cataaaatag gaactaaaat
      361 accaaaatag aaacatttat atacatttga tgtgtagggt ataccataaa cctgggaggt
      421 gtctcctatg gtaaagtatg aagataagat aagtttgtac gatgcgaaag gaaacctagt
      481 agccgaaaat gtaccattag aagctattag cccattatac aaccctacca taaaaagtat
      541 ggtaaagaac atcaaaagaa cagttgctgt gaacctagct ggtatcgaag gtaccttagc
      601 tgcaggtaaa atcggtggaa aaggctgtca agttccagga agaactttag atatatctgc
      661 agtaagcaat gctcaagcaa ttgctgacga agtagaaaaa atcttaaaag tatccgaaga
      721 cgacgataca gcagtgaaaa taatcaacgg tggaaaacag ttagctgtgc aagttccaac
      781 cgcaagattg gacgtagctg cagaatactc agtatccatg ctttcaactg ctatggcatt
      841 aaaagaagca cttattaaaa cattcaacat cgacatgttc gatggttcaa cagtgcacgc
      901 tgcaattgtt ggtaactacc cacaggttat ggattacgca ggcggtaaca ttgcatcact
      961 cttaggtgca ccttcaatga tggaaggttt aggttacgct ttaagaaaca ttccagttaa
     1021 ccacgcagtt gctacaacca agaaaaacat gatgaacgct attgcattct catcagttat
     1081 ggaacaaact gcaacattcg aaatgggtga cgcagtagga tcattcgaaa gacagcactt
     1141 acttggttta gcttaccaag gattaaacgc tgataactta gttattgact ttattaaagc
     1201 aaatgcaaaa ggaactgttg gttcagttgt agaaacagtt attgatagag ctattgctga
     1261 cggcgtaatc gttgttgaca aaacaatgag ctcaggattc aacatgtaca aaccagctga
     1321 cgtaaacaaa tggaatgcat acgcagcagc aggattagtt gcagcagttg cagtaagctg
     1381 tggtgcagca agagcagcac agaacgttgc ttcagttatt ttatacttca acgatatttt
     1441 agagtacgaa actggtttac ctggtgttga ctacggtaga tcaatgggta ctgcagtagg
     1501 attctcattc ttctcacact caatctacgg tggtggtgga cctggtatct ttaacggtaa
     1561 ccacgttgta accagacact ccaaaggatt cgcaattcct ccagtatgtg cagcaatgtg
     1621 tgcagatgca ggtacacaaa tgttctcacc tgaacacact tcaggattag ttggttcagt
     1681 atactcagca tttgatgaat tcagagaacc aatgaaatac gtaattgaag gagctttaag
     1741 cattaaagac cagttttaag gtaataaacg atgattgaat tagaagtctt cccacatagg
     1801 tatttaaaag cagaaactac cgaaaaattc ctgaaccgcg cttattcatt gagtaaagtt
     1861 caaagagtag tcatacatgg agaatctctt cctaataaag ttggatacgg tccagctaaa
     1921 ggaactcctg taaatcactc agaaaaaaag gaaattactg ttaagggagt ccctgtagaa
     1981 ctcatgcttc aagttggaag gttatgggtt ttactggatg atgaaagcga aattgaaaaa
     2041 atcgaagaaa tctgtaagga tcttttccca tttggatacc ggttaacaaa gggtaaattc
     2101 ttaaggaacg accctacagt atcggatttc ataaaatacg gggaatctgc tgttgatgat
     2161 atcgacaaaa gattattagg tgcgactgac cccagaagta agtttgattc ttctgtcaca
     2221 attatcccaa agagcgaaaa aaacgagtaa gtgattttat gccagtaggt agaaaagagc
     2281 agatcgttga ttgtagagct gttatgggtc ttggtgaagg aggcggactt gcacaaagag
     2341 gaacatttgc agaagggcta cgtaacgacg tggttgttgt tgcaatgtct ccagggagaa
     2401 ggcacattac aaaacccgtg tgcgaaatta cctatggaat aagggaagcg ggaattcaga
     2461 ccagcgttct tgtattagat gcaggaggag gaattccgtc agatgctcca caaggaagtt
     2521 tgggttccac ttttggactt aaacccgaag aagctaaaca agttaatagg cataaattat
     2581 gtgtaataca ttttggaaat gtaaaaagtc acataattta caaagctaga ttattcttaa
     2641 aatatgtgga tattcctaca ataattgttt gccaaactcc agtagatatg gaagatttcg
     2701 cagcaattgg aataaaaacg aaaaatgtaa tgcctttgga atcaaaaact gaaggtaaaa
     2761 tcgttgaaat tattactggc gtaataagag gagaatcagc tcctcaaaaa aaaattgatg
     2821 aaatcattga aagcataaaa aaacatttag gttaaggtga tcttatggat tacaaagatg
     2881 atgatgataa aggcgccagc tggtcacacc cacaatttga aaaaggaggt ggatcaggag
     2941 gtggatcagg aggttcagca tggtcacatc ctcagtttga aaaatcagca atggcataca
     3001 cgcctcagtt ctaccctggt gcaacaaaag ttgctgaaaa cagacgaaat cacttgaatc
     3061 caaactatga attggaaaaa ttaagagaaa tccctgatga agatgttgta aaaatcatgg
     3121 gacacagaca acctggtgaa gactacaaaa cagttcaccc accacttgaa gaaatggact
     3181 tcgtagaaga ctacgcaaga gacttagtag aaccattaaa tggtgctaaa gaaggacaca
     3241 gagttagata cattcagttt gctgactcaa tgtactttgc tcctgctcaa ccatacgaca
     3301 gatcaagatc atacatgaca agattaagag gagtagatgc aggtacactc tcaggaagac
     3361 aagttgttga atgtagagaa agtgaccttg aagaattctc caaaaacata ctcatggata
     3421 cagaactctt tgaccctgct acatcaggtg ttaggggtgc aaccgtacac ggacactcat
     3481 taagattaga tgaaaacggt atgatgtttg atgcactcca aagatgtgta ttcgacgaaa
     3541 aaaccggaca cgttatgtac gtaaaagatc aggtaggtaa accactcgat gcacctgttg
     3601 acgttggaga accaatccct gaagcaaaat taagagaaat tacaaccatc tacagaaacg
     3661 atggtgttgc aatgagagca gatcctgacg ttatcgaagt tgtaaaaaga attcacagag
     3721 caagaacact cggtggatac atccctacaa acgaaacatt caaaggatta taactaaatt
     3781 ttaaattaaa aaaaacctct taaaccattg aggtgaactt atggaagctg aaaaaagatt
     3841 atttttgaaa gccttaaagg aaaaattcga agaagaccct aaagaaaagt acacaaaatt
     3901 ctataccttt ggcggttggg aacaatctgc tagaaaaaga gaatttgttg aagctaatga
     3961 aaaaatagta tctgaaaaaa gacaaggaat cccactctac aacccagaca ttggtgttcc
     4021 tctaggacag agaaaattaa tgccttacaa attgtcaaac actgacgatt actgtgaagg
     4081 tgacgattta cacttcttaa acaacgctgc tatccaacaa ttatgggacg atatcagaag
     4141 aactgttatt gtaggtatgg acactgctca ctcagtttta gaaaaaagat tgggtgtaga
     4201 agtaactcct gaaacaatca acgaatacat gcacacaatc aaccactccc ttccaggagg
     4261 agctgttgtt caggaacaca tggttgaagt tcacccttcc cttgcatggg actgttacgc
     4321 tagaatcttc accggtgacg atgaattagc tgacgaatta gacagcagat tcttaatcga
     4381 catcaacaaa ttattccctg aagaacaagc tgaaacatta aaagctgcaa ttggtaaaaa
     4441 aacataccaa gtatcaagag ttccatcact cgtaggtaga gtatgtgacg gcggtacaat
     4501 ctcaagatgg tctgcaatgc agatcggaat gtcattcatt accgcataca aactttgtgc
     4561 aggggaagct gcaacagctg acttctcata cgcttcaaaa cacgctgatg taattcagat
     4621 gggtaatgct ttaccaggaa gaagagcaag aggtccaaac gaaccaggag gaatcagatt
     4681 cggtattctc tccgatgttg tacaaacaac aagagtttca gaagacccag ttgaacaatc
     4741 attagaagta gtagcaacag gtgctgcttt atatgaccag atctggctcg gtgcatacat
     4801 gtctggtggt gtaggattca ctcaatacgc taccgcatca tacaccgacg atatcttaga
     4861 tgacttctca tactacgcac tcgactacgt tgagaaaaaa tacggtagaa tgggaacaaa
     4921 agctacaatg gatgtagtag aagatgttgc tggagaagtt acactctacg cattagaaca
     4981 gtacgatgac tacccagcat tattagaaga ccacttcggt ggatcacaaa gagctgcagt
     5041 tgctgctgct gcatcaggta ttggtgtatg tatggcaaca ggaaactcaa acgctggtgt
     5101 aaacggctgg tacttatcac aaatcttaca caaagaatac cacagcagac tcggattcta
     5161 cggttacgac ttgcaggacc agtgtggtgc ttcaaactca ctcgcaatca gaaacgacga
     5221 agctgcacct cttgaattaa gaggacctaa ctaccctaac tacgcgatga acgtaggtca
     5281 ccagggagaa tacgcaggta ttgcacaagc tgcacactcc gcaagaggag acgcatttgc
     5341 acttagccca ttggtaaaag ttgcatttgc tgacccaatg ctcgtattcg atttctccaa
     5401 acctagaaaa gaaatggcta ggggagcatt aagagaattc gaagctgcag gagaaagaga
     5461 cgttatctta cctgctaaat aaactagtag cggccgctgc aggatatatt aaaagtggga
     5521 cttaagttcc cacttttttt attttataaa aaacgcccta ttcgtaatat actattcaaa
     5581 actaaatatt cattgaaata tttgggtaat aattaatatt aatctaatat ttagatattg
     5641 agatataata ataacatttt tttaaaaaat catctatttt catcttaatt ttatatatta
     5701 aagcccattt tttgagtatt caaattcaaa ttattgtgtt attaacatct tatatataaa
     5761 cttttctatt taatgttaat gaaaaagtga atatatatac atagagtaat gttatgatgt
     5821 atatatcaaa aaaaactaga ggagacgacc ttccatgaca gaatataaac caacagttag
     5881 attagcaaca agagatgacg tacctagggc agtaagaact cttgcagcag cttttgcaga
     5941 ttatccagca acaagacaca cagttgatcc agatagacac attgaaagag ttacagaatt
     6001 acaagaatta tttttaacaa gagttggtct tgatataggt aaagtatggg ttgcagatga
     6061 tggagcagca gttgcagttt ggacaacacc agaatcagtt gaagctggtg ctgtttttgc
     6121 agaaattgga ccaagaatgg ctgaattatc aggatcaaga ttagcagctc agcaacagat
     6181 ggaaggatta cttgcaccac acagaccaaa agaaccagca tggtttttag caacagttgg
     6241 agtttcacca gatcaccagg gaaaaggttt aggttctgct gttgtacttc ctggtgttga
     6301 agcagctgaa agagcaggag ttccagcatt tttagaaaca tcagcaccaa gaaatcttcc
     6361 attttatgaa agacttggat ttacagttac agcagatgtt gaagttccag aaggaccaag
     6421 aacatggtgt atgacaagaa aaccaggagc ataaccatgg ctccgaccga agccacccgg
     6481 ggcggccccg ccgaccccgc acccgccccc gaggcccacc gcgggggaca caccgaacac
     6541 gccgaccctg ctgaacacgc ggcgcagttc ggtgcccagg agcggatcgg gaattaattc
     6601 gaagctgctg gtgaaagaga ccctatctta cctgctaaaa tctaagttaa ttactaattt
     6661 attattaatt tattattaga ttgggtaaaa tagtaaaaga aaactaaagg aaacctaata
     6721 tggtttcctt tttttatata ttttaattca ctgggggtga tttaaaactt catttttaat
     6781 ttaaaaggat ctaggtgaag atccttggtg cactctcagt acaatctgct ctgatgccgc
     6841 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6901 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6961 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     7021 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7741 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7801 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8041 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg tacccattat
     8881 ttagtttttt tttatcacca caacgtataa accccctttc agaagaaaac agagaatgaa
     8941 aacagatttt aatttatata tcgtatctat aatttagttt ttttttattg cagttttaga
     9001 agtgcgtttg tcctggagcg aaatacacga tatcgtgtag tctccttcag atgaaaataa
     9061 tatttttaat tttcagaaat attactggaa aaatacctga aaaatatttg aaaaaaaaac
     9121 atacacccag cggggaaaaa tcgaacattt tgaaaaagtc tatttatagg gtttaaacag
     9181 ggccaagacc tgcgggatat accccaaaaa tgttcgataa aaaaatcgaa catttttcga
     9241 acatttttta taatgttcga tgtgtgcgag tatgttcgat aaaaaaatcg aacattttga
     9301 aagacaattt gaagagatat aaaaaaaata atctattata atatagatat ataatatgtt
     9361 cgatatgttc gatatttttt tatatattat attatattta accttgtatt gattat
//
