LOCUS       sequence-379179-        8391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged ANME1BS MCR in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8391)
  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 8391)
  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-ANME1BS MCR-NG
FEATURES             Location/Qualifiers
     source          1..8391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2263..2286
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2296..2319,2320..2355,2356..2379)
                     /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(6307..6326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6426..6448
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6486..6504)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6572..6676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6677..6745,6746..7537)
                     /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(6895..6914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7708..8296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8197..8216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtataaaccc cctttcagaa gaaaacagag aatgaaaaca gattttaatt tatatatcgt
       61 atctataatt tagttttttt ttattgcagt tttagaagtg cgtttgtcct ggagcgaaat
      121 acacgatatc gtgtagtctc cttcagatga aaataatatt tttaattttc agaaatatta
      181 ctggaaaaat acctgaaaaa tatttgaaaa aaaaacatac acccagcggg gaaaaatcga
      241 acattttgaa aaagtctatt tatagggttt aaacagggcc aagacctgcg ggatataccc
      301 caaaaatgtt cgataaaaaa atcgaacatt tttcgaacat tttttataat gttcgatgtg
      361 tgcgagtatg ttcgataaaa aaatcgaaca ttttgaaaga caatttgaag agatataaaa
      421 aaaataatct attataatat agatatataa tatgttcgat atgttcgata tttttttata
      481 tattatatta tatttaacct tgtattgatt attgaataat atttattatt actttttttg
      541 atttcttgta aaaaatcgaa catattttac atattgtaaa aaataaaaaa gatttaaaaa
      601 aagtgtccga aaaagttaaa caaacattat tattaaaatg ttagtttaat atgtatcttg
      661 atatgctctt ctttctttca tgcttaaact tgtataactg tatgcttttt tcatttcctt
      721 tattttgtcc tttcctttct ttgttatctg gtaatatcct tccttctttc ctgcatactt
      781 caaaaatcca aggtttttta agttatcgag ataagaatta ctagatgaat tcgcgcccgg
      841 ttctagattt attcataaaa taggaactaa aataccaaaa tagaaacatt tatatacatt
      901 tgatgtgtag ggtataccat aaacctggga ggtgtctcat atggcagatg aaattgatct
      961 ttacgatgat agaggatcag ttttagcaaa aggagttcca cttgaagcaa tttcaccatt
     1021 aaaaaacgca gcaattagaa aaattgtttc aatgacaatt agaactggag cagttgatat
     1081 ggcaggactt gataaaaaac ttaaaacagg aactattgca ggaagaggaa tggttattag
     1141 aggagttgga agagatattc cagttcttga aaaagttaac gaaattgcaa aagaagttga
     1201 agatatgctt agagttgaac caggagatga tacaaaagtt gaacttcttg caggtggaaa
     1261 aagaatgctt acacaagttc caactgcaag aattctttca gattactcag ttggattaac
     1321 tgcagcaatg ggagcattaa cacacgcaat tattgatgtt tgtgatgttt caatgtggga
     1381 tgcaccatat gttcacgcag cagtttgggg aatgtaccca cagaatccag atccagcaga
     1441 tggagcagtt aaaatgttag ttgatattcc aatgaaaaac gaaggaccag gatttactct
     1501 tagaaacatt ccagttaacc acttagcagc aacagttaga aaaagagcaa ttcaaggatc
     1561 agcacttact atgattttag aagaagcagc acagtttgaa atgggaaatt gtatgggacc
     1621 acacgaaaga ggacacttat tagatttagc atatgaagga ttaaatgcaa acaacattct
     1681 ttacaacctt attaaagata atggaaaaac aggaactctt ggagatgttg tttactcatt
     1741 agttgataaa gcaaaagcag atggagttat taaacctaaa aagaaaatgg gatcaggata
     1801 tacagtttac actgcagatg atccacaatt atggaacgca tacgcatcag caggattatt
     1861 agcagcagtt tgtgttaatt gtgcagcaat gagagcagga cagtcagttc caggatgtat
     1921 tatgtactac aacgttcttc ttgaacacga aacaggaatg ccaggagttg ataatggaat
     1981 ggcacaagga gcatcagttt catcatcatt tttctcacac tcaatttatg gaggcggagg
     2041 accaggagta ttttatggaa accacattgt tactagacac ccaaaaggac agtttattcc
     2101 atgtttttgt gcatcaatgt gtcttgatgc agatacaatg tactttactc cagcaagaac
     2161 atcagcatta tacggagaag ttttaggagc aattccagaa tttgcagaac caatgaaagc
     2221 ggttgcagga gccgtataaa taaggaggaa agaaaaaaaa tggattacaa agatgatgat
     2281 gataaaggcg ccagctggtc acacccacaa tttgaaaaag gaggtggatc aggaggtgga
     2341 tcaggaggtt cagcatggtc acatcctcag tttgaaaaat cagcaatgcc acaatattgt
     2401 cagggaaaaa cacacattgc agatactaga agaaaataca tgaacccaga tgttaaactt
     2461 gaaaaactta gagatattgc agaagaagat attgttagac ttcttgcaca cagagcacca
     2521 ggagaagaat ataaatcaat tcacccacca ttagaagaaa tggaagaacc agattgtgca
     2581 gttagacaaa tggttgaacc aacagaagga gcaaaagcag gagatagaat tagatacgtt
     2641 caatacacag attcaatgtt tttctcacca attactccat atttaagagc acagtcagca
     2701 ttttgtagat acagaggaat tgatccagga gttctttcag gaagagttat tattgaatca
     2761 agagaaagag atgttgaaaa aattgcaaaa gaaacaattg attcagaatt atacgataca
     2821 gcaagaactg gattaagagg aagaactgtt cacggacacg cagttagatt agatgcaaat
     2881 ggaatgatgt ttgatgcact tagaagatgg tcaattgatg aaagaacaaa agaagttact
     2941 tacgttaaag atatgattgg aggagcaatg gataaagaag ttgttcttgg aaaaccactt
     3001 ccagaagcag atttattaaa aagaacatgt atgtacagaa atgcacaggg aggagtttgg
     3061 caggaagaag atgatccaga atcaatggat gtttgtgcag aaattcactg gaaaagaact
     3121 gttggaggat ttcagccatg gaagaaaatt tcagaaatta aaggtggaaa gaaagatatt
     3181 ggagttaaaa acctgaagtt attcgtacca agaggaggag tattataaaa tgccatacaa
     3241 cgatattcaa cactcattta tgaaagcaat gtcagataaa tttgcagaaa aaccagaatc
     3301 aacagaaact aaattttacg tttacggagg aattgcacaa aaaggtggaa tgagaaaaag
     3361 agaatttatt gcagaagcac agaaaattgt tgattcaaga gtttcaggaa caccagcata
     3421 taatccagat gttggaatgc cacaaggaca gagattttta atgccataca tgatgaacca
     3481 cactgatatt atggttaacg cagatgattt acactggatt aacaatgcag caatgcaaca
     3541 gtgttgggat gatatgaaaa gaggaattgt tcttggactt gatgatgcac acggacttct
     3601 tgaagcaaga cttggaaaag aagttacacc agatactatt tcacactaca tggaagttct
     3661 taatcacgca ttacctggag gagcagttat tcaggaacac atggttgaaa caaaaccaat
     3721 gcttgttaac gattcatacg caaaaatttt tactggagat gatgatcttg cagatgcagt
     3781 tgatagaaga tttttacttg atattaacaa agaatttgca gcaggatggg aaaaaccagg
     3841 agaacaagca gatcagctta aagatgcaat tggaaagaaa atttggcaaa ttctttggat
     3901 gccaacagtt gttggaagac agacagatgg aggaactatg tttagatggg ttggaatgca
     3961 agttggaatg acaatgattt cagcatacaa actttgtgca ggagaatcag ttactggaga
     4021 atttgcatat tacgcaaaac acgcagcagt tgttcagctt tcaaactaca tgccagttaa
     4081 aagagcaaga gcacacaatg aacctggagg aatgccatta ggaattaatg cagattcagt
     4141 tagatcacca gcattatttc caaatgatcc aattagagca gaattagaat caattgcagt
     4201 tgcagcaatg gtttacgatc aactttggtt tggaacatac atgtctggag gagttggatt
     4261 tactcagtac gcatcagcaa catacactga taacattctt gaagattttt gttacaaagg
     4321 atgtgaaatt ggattagatt acgcaaatgg agaaatggga tcaattaaag gagataaact
     4381 taacatggat attcttgaag aaatgattag agcagaaaac gattactgtc ttacacaata
     4441 cgaagcatac ccaactgttg cagaatcaca ctttggagga tcagttagag catgttgtgc
     4501 agcagcagga tgtggatcaa cagttgcatg tgcaactgga ttagcacagc caacattatc
     4561 agcatggtca ctttcaatgc ttggacacta cgaaagaaaa ggaagattag gatttttcgg
     4621 atacgattta caagatcagt gtactgcatg tggatcatat tcataccaat cagatgaagg
     4681 aatgccattt gaaatgagag gagttaacta cccaaactac gcaatgaacg ttggacacca
     4741 gtcagcatat gcaggattag ttatgggagc acacgttgca aataaagacg catgggttct
     4801 ttcaccatta tggaaagttg cattttcaga tagagattta ccatttgata gaggatatgt
     4861 tacaagagaa tacggattag gagcaaatag agaatacact aaagttgcag gagaaagaga
     4921 tcttattatt gcaggacact atggaagaga accaggagca aaattataaa ctagtagcgg
     4981 ccgctgcagg atatagcaaa agtgggactt aagttcccac ttttgctatt ttataaaaaa
     5041 cgccctattc gtaatatact attcaaaact aaatattcat tgaaatattt gggtaataat
     5101 taatattaat ctaatattta gatattgaga tataataata acattttttt aaaaaatcat
     5161 ctattttcat cttaatttta tatattaaag cccatttttt gagtattcaa attcaaatta
     5221 ttgtgttatt aacatcttat atataaactt ttctatttaa tgttaatgaa aaagtgaata
     5281 tatatacata gagtaatgtt atgatgtata tatcaaaaaa aactagagga gacgaccttc
     5341 catgacagaa tataaaccaa cagttagatt agcaacaaga gatgacgtac ctagggcagt
     5401 aagaactctt gcagcagctt ttgcagatta tccagcaaca agacacacag ttgatccaga
     5461 tagacacatt gaaagagtta cagaattaca agaattattt ttaacaagag ttggtcttga
     5521 tataggtaaa gtatgggttg cagatgatgg agcagcagtt gcagtttgga caacaccaga
     5581 atcagttgaa gctggtgctg tttttgcaga aattggacca agaatggctg aattatcagg
     5641 atcaagatta gcagctcagc aacagatgga aggattactt gcaccacaca gaccaaaaga
     5701 accagcatgg tttttagcaa cagttggagt ttcaccagat caccagggaa aaggtttagg
     5761 ttctgctgtt gtacttcctg gtgttgaagc agctgaaaga gcaggagttc cagcattttt
     5821 agaaacatca gcaccaagaa atcttccatt ttatgaaaga cttggattta cagttacagc
     5881 agatgttgaa gttccagaag gaccaagaac atggtgtatg acaagaaaac caggagcata
     5941 accatggctc cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag
     6001 gcccaccgcg ggggacacac cgaacacgcc gaccctgctg aacacgcggc gcagttcggt
     6061 gcccaggagc ggatcgggaa ttaattcgaa gctgctggtg aaagagaccc tatcttacct
     6121 gctaaaatct aagttaatta ctaatttatt attaatttat tattagattg ggtaaaatag
     6181 taaaagaaaa ctaaaggaaa cctaatatgg tttccttttt ttatatattt ttaattcact
     6241 ggggggattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc cttggtgcac
     6301 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     6361 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6421 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     6481 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6541 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6901 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8041 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8101 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8161 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8221 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8281 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8341 gccttttgct cacatggtac ccattattta gttttttttt atcaccacaa c
//
