LOCUS       sequence-379187-        8520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged E50 MCR in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8520)
  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 8520)
  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-E50 MCR-NG
FEATURES             Location/Qualifiers
     source          1..8520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1540..1563
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1573..1596,1597..1632,1633..1656)
                     /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(5603..5622)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5722..5744
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5782..5800)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5868..5972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5973..6041,6042..6833)
                     /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(6191..6210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7004..7592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7493..7512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatgaattc gcgcccggtt ctagatttat tcataaaata ggaactaaaa taccaaaata
       61 gaaacattta tatacatttg atgtgtaggg tataccataa acctgggagg tgtctcatat
      121 ggcatacctt actgaaaaaa ttgatcttta cggagataac ggaaaagttc ttgaatcaga
      181 tattccatta gaagcagtta caccagttca aaatccagca gttagagaac ttgcatcaat
      241 ttttaaaaga tcagttgcag ttaatcttgg aggagcacag aaagcattat caactggaca
      301 ctatgcaaat gaatacattc actttccaga tattccaaac aaagataaac ttggaattaa
      361 atcatcacct ggaggaaaat atccaccaaa atcagttaaa gttagaacta tggatttacc
      421 attagttgat gatgcagatg atattgcagc aagacttaaa gaaagacttc aagttaatcc
      481 agatgatgga acagaagtta gagttatgaa gaaaggaaac gttctttacg ttaaaatttc
      541 agaacaatta gcaaatactg gagttgaata cacaactgca ttaacaacta cagcacaggc
      601 aatgacagat cttgttatgg aaaaatacga tcttgatttt cacgcatcac cattagttca
      661 ctgtgcattt tatggaagat acccacagac ttacgaattt atgggaggaa acgttatttc
      721 acttcttgca gcatcatgtg caaatgaagg accaggattt gcaatgagaa acattatggc
      781 aaaccacatt gttgcagcaa ctagaaaaag aacattagaa gcagttgcat tatcatcaac
      841 attagaagca attggacacg ttgaaatggg agatgcaatt ggaagatgga gaagatggca
      901 agcattagtt cacgcatgtc agggacttaa cgcaaacaac gttgtttacg atttagttaa
      961 agaagcagga cacggatgta ctggagatgt tgttgcagca acagttggaa gagcacttga
     1021 agatggaatt atttcagtta agaaaacact tccatcagga tacaaatttt acacagcaaa
     1081 cgatccatca atgtggaatg catacgtttg tgcaggatta gttgcagcag ttattgttaa
     1141 tcaaggagca gcaagagcag cacagggagt ttcatcaact cttctttact ttaacgatct
     1201 tattgaacac gaaacaggat taccacacgc aggatacgga gatggaatgg gaaacggagt
     1261 ttcattttca tttttctcac acgcaattta tggtggagga tcaccaggaa tattttcagg
     1321 aaaccacatt gttacaagac actcaaaagg atttgcaatt ccagttattg cagcagcagt
     1381 ttcattagat tcaggaactg cagtttacgg accagaagca acatcaggat tagttggaga
     1441 tatttttgga gaagttgatc ttattagaag accaatggaa gcgatcgcaa gtgcagcagc
     1501 agagataaag gacaagttct gaagaggtga agaagaatgg attacaaaga tgatgatgat
     1561 aaaggcgcca gctggtcaca cccacaattt gaaaaaggag gtggatcagg aggtggatca
     1621 ggaggttcag catggtcaca tcctcagttt gaaaaatcag caatggttta ccaaagacag
     1681 tttttaccag cagatgatag agttacaaaa aacagaaaga aagttgttga tccatcagtt
     1741 aaacttgaaa aaattagaac attatcagat aaagattttc ttactttaat tggacacaga
     1801 cacttaggag aagcatatag atcagttaat ccaccattag cagaaattgg agaaccagaa
     1861 gatccaatta gagaattagt tccaccaact gaaggagcaa aagcaggaga tagagtttgt
     1921 acaattatta tgactgattc agtttacaat ccaccaattg cacactatac aagagcatgg
     1981 atgtaccaca acagatttag aggaattgat aacggagttt actcaggaag agttactctt
     2041 gaaatgagag aaagagatct tgaagaagca tgtagaacac tttttgaaac tgaaatttgt
     2101 gatgcatcaa gagatcaagt tagacagtat acatgtactg gacactcatg tagattagat
     2161 ccagatggaa tgatgtttga tccaattgaa agatgtatta tgtctggagg aaacgttgtt
     2221 taccaaaaag attcatttgg aaacccagtt gatacaccaa ttaacatggg aaaaccactt
     2281 tcagaagaag aacttattga aagaacagtt gtttacagaa ctgatagagg agaaccaatg
     2341 actagagaag gagatccagg agcaccagat gaagaagtta gagaagcatt acaatggtca
     2401 agaagaattc agtggctcag aatgctcggg aacatggttc ctgacaagat aaagggaatg
     2461 taaagaggtg aaatgaaatg gtaaaatatc caaagcaatt gttcttagag agcaagaact
     2521 caaagatgaa ttcaattgaa atgaaatatg gacaagatcc agcaattaat agagcagaat
     2581 ttcacgttta cggaggagtt agacagtcaa aaagaaaatc agaagcatgg gaagcagcaa
     2641 aaagaattac aaaagaaaga ggaattccaa actacaaccc agatttacac ttaaaaggag
     2701 cacaaatggg acagaaagtt ttacagacat acagaattac tggattagat agagaatggg
     2761 caggtggaga agatacacca gcacacaaag gatggaaacc aggaactgat attgcaggac
     2821 ttgaaatgga tgatcttaac tacgaaaata atccagcaat gcaacagtgt tacgatgata
     2881 tgagaagaac agcaattaac ggactttcaa ttgcacacga aactattgaa agaagatttg
     2941 gaaaagaagt tacaccagaa actattaacc tttactttga aatgcttaac cacaacattg
     3001 gagcaggagc aattatgatg gaacacacag cagaaactaa tccagaactt gttaaagatt
     3061 catacgcaaa atgttttact ggaaacgatg aattagcaga tgcattagat caaagatttt
     3121 taattgatat taacaaaatg tttccaaaat accaagcaga tcagattaaa gcagaagttg
     3181 gagatagaat ttttcaggtt gcaagaattc caacaatggc agttagaact tcagatggag
     3241 gattatcaag agcatgggtt ggacaacagg catcacttgc atttttatgt gcatacgata
     3301 ttccagcagg agatgcagtt acatcagatt ttgtttttac tattaaacac ggagatgttg
     3361 tttttatggg aacacaatta ccatacagaa gagcacagag aaataattca gcaggtggaa
     3421 ttgcacttgg atactactca gattgtaacc aaacatcaag aactccagaa gcattagaag
     3481 gattagatgg aggaattgat ccagttaaag ttattgttga agcacttaca ccaggatgtg
     3541 ttattactga tcagggatgg ttacacaatt atttagcagg tggatcatca ggatggtcaa
     3601 actacattat ttcagtttac actgatgaag ttcttgaaga ttatggatac cacggagcaa
     3661 tttatgcaat ggataaatgg aaatgtggag ttggagaagt tccaaacaca tacgaaaaca
     3721 tgatgactat tgcagaagaa gtttcaagat ggtcacagaa aaattacgat gaatacccag
     3781 gacttatgga agcacacttt ggaggatcac aaagatattc aattcaggca gcagcatcag
     3841 gagcagcagt tggagcaatg acaggagatc cagatttagg aaatgcagca tggcactaca
     3901 atactccatt atgtaaagaa cactatttaa gacttggatt ttacggacac gatttacaag
     3961 atcaacagaa tatgggacac acatattcat acagatcaga tcagggaatt ccatacgaac
     4021 ttaaaggacc aaactaccca gattttgcaa tgaatgttgg acacatggga ggatatattg
     4081 gaattattgc aggagcagca cacgcaagag gagcagcata ctcaactaac ccaattatta
     4141 aagcagcatt tgcagatcca aatttacaat ttgattttag atatccaaga agagaatttg
     4201 gaattggagg attaagacag tttatgccag caggagaaag agatgcagtt attccaccac
     4261 actaaactag tagcggccgc tgcaggatat agcaaaagtg ggacttaagt tcccactttt
     4321 gctattttat aaaaaacgcc ctattcgtaa tatactattc aaaactaaat attcattgaa
     4381 atatttgggt aataattaat attaatctaa tatttagata ttgagatata ataataacat
     4441 ttttttaaaa aatcatctat tttcatctta attttatata ttaaagccca ttttttgagt
     4501 attcaaattc aaattattgt gttattaaca tcttatatat aaacttttct atttaatgtt
     4561 aatgaaaaag tgaatatata tacatagagt aatgttatga tgtatatatc aaaaaaaact
     4621 agaggagacg accttccatg acagaatata aaccaacagt tagattagca acaagagatg
     4681 acgtacctag ggcagtaaga actcttgcag cagcttttgc agattatcca gcaacaagac
     4741 acacagttga tccagataga cacattgaaa gagttacaga attacaagaa ttatttttaa
     4801 caagagttgg tcttgatata ggtaaagtat gggttgcaga tgatggagca gcagttgcag
     4861 tttggacaac accagaatca gttgaagctg gtgctgtttt tgcagaaatt ggaccaagaa
     4921 tggctgaatt atcaggatca agattagcag ctcagcaaca gatggaagga ttacttgcac
     4981 cacacagacc aaaagaacca gcatggtttt tagcaacagt tggagtttca ccagatcacc
     5041 agggaaaagg tttaggttct gctgttgtac ttcctggtgt tgaagcagct gaaagagcag
     5101 gagttccagc atttttagaa acatcagcac caagaaatct tccattttat gaaagacttg
     5161 gatttacagt tacagcagat gttgaagttc cagaaggacc aagaacatgg tgtatgacaa
     5221 gaaaaccagg agcataacca tggctccgac cgaagccacc cggggcggcc ccgccgaccc
     5281 cgcacccgcc cccgaggccc accgcggggg acacaccgaa cacgccgacc ctgctgaaca
     5341 cgcggcgcag ttcggtgccc aggagcggat cgggaattaa ttcgaagctg ctggtgaaag
     5401 agaccctatc ttacctgcta aaatctaagt taattactaa tttattatta atttattatt
     5461 agattgggta aaatagtaaa agaaaactaa aggaaaccta atatggtttc ctttttttat
     5521 atatttttaa ttcactgggg ggatttaaaa cttcattttt aatttaaaag gatctaggtg
     5581 aagatccttg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5641 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5701 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5761 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5821 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5941 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6001 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6061 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6121 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6181 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6241 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6301 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6361 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6421 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6481 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6541 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6601 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6661 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6721 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6781 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6841 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6901 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6961 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7021 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7081 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7141 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7201 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7261 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7321 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7381 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7441 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7501 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7561 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7621 tcctggcctt ttgctggcct tttgctggcc ttttgctcac atggtaccca ttatttagtt
     7681 tttttttatc accacaacgt ataaaccccc tttcagaaga aaacagagaa tgaaaacaga
     7741 ttttaattta tatatcgtat ctataattta gttttttttt attgcagttt tagaagtgcg
     7801 tttgtcctgg agcgaaatac acgatatcgt gtagtctcct tcagatgaaa ataatatttt
     7861 taattttcag aaatattact ggaaaaatac ctgaaaaata tttgaaaaaa aaacatacac
     7921 ccagcgggga aaaatcgaac attttgaaaa agtctattta tagggtttaa acagggccaa
     7981 gacctgcggg atatacccca aaaatgttcg ataaaaaaat cgaacatttt tcgaacattt
     8041 tttataatgt tcgatgtgtg cgagtatgtt cgataaaaaa atcgaacatt ttgaaagaca
     8101 atttgaagag atataaaaaa aataatctat tataatatag atatataata tgttcgatat
     8161 gttcgatatt tttttatata ttatattata tttaaccttg tattgattat tgaataatat
     8221 ttattattac tttttttgat ttcttgtaaa aaatcgaaca tattttacat attgtaaaaa
     8281 ataaaaaaga tttaaaaaaa gtgtccgaaa aagttaaaca aacattatta ttaaaatgtt
     8341 agtttaatat gtatcttgat atgctcttct ttctttcatg cttaaacttg tataactgta
     8401 tgcttttttc atttccttta ttttgtcctt tcctttcttt gttatctggt aatatccttc
     8461 cttctttcct gcatacttca aaaatccaag gttttttaag ttatcgagat aagaattact
//
