LOCUS       sequence-379057-        9512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-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-NG
FEATURES             Location/Qualifiers
     source          1..9512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2978..3001
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(3011..3034,3035..3070,3071..3094)
                     /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(6951..6970)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7070..7092
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7130..7148)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7216..7320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7321..7389,7390..8181)
                     /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(7539..7558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8352..8940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8841..8860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatattata ttatatttaa ccttgtattg attattgaat aatatttatt attacttttt
       61 ttgatttctt gtaaaaaatc gaacatattt tacatattgt aaaaaataaa aaagatttaa
      121 aaaaagtgtc cgaaaaagtt aaacaaacat tattattaaa atgttagttt aatatgtatc
      181 ttgatatgct cttctttctt tcatgcttaa acttgtataa ctgtatgctt ttttcatttc
      241 ctttattttg tcctttcctt tctttgttat ctggtaatat ccttccttct ttcctgcata
      301 cttcaaaaat ccaaggtttt ttaagttatc gagataagaa ttactagatg aattcgcgcc
      361 cggttctaga tttattcata aaataggaac taaaatacca aaatagaaac atttatatac
      421 atttgatgtg tagggtatac cataaacctg ggaggtgtct catatggtta aatacgaaga
      481 taaaattaac ctttacgatg caaaaggaaa ccttgcagaa gaaaacgttc cacttgaagc
      541 aatttcacca ttatacaatc cagttgttaa agatatggtt aaatcaatta aaagaacagt
      601 tgcagttaac cttgcaggaa ttgaagcaac acttaaagca ggaactattg gaggaaaagc
      661 atgtagagtt ccaggaagaa ctcttgattt accaattgtt gatgaagcag aagcaattat
      721 ggataaagtt gaaaaaattc ttaaaatttc agaagatgat gatacaaata ttagaactat
      781 taatggagga aaacaagcag ttgttcaggt tccatcaaaa agacttgata ttgcagcaga
      841 atactcagtt tcaatgttat caacaggaat ggcacttaaa gaagcaatta ttgatacatt
      901 tgatgttgat atgtttgatg gatcaactgt tcacgcagca attgttggaa gatatccaca
      961 agttatggat tacatgggag gaaatattgc atcattatta ggagcaccat caaatatgga
     1021 aggattagga tacgcactta gaaacattat ggttaaccac tacgttgcaa caacaaagaa
     1081 aaatcttatg aacgcagttg cattttcatc aattatggaa caaactgcaa tgtttgaaat
     1141 gggagatgca attggatcat ttgaaagaat gcacttatta ggacttggat accagggact
     1201 taacgcagat aacattgtta ttgatttagt taaagcaaat gcatcaggaa cagttggaac
     1261 tgttgttgca tcagttgttg aaagagcact tgaagataaa gttattgcag aagataaaac
     1321 aatgaactca ggatttacta tgtacaaacc agttgatgtt gcaaaatgga atgcatacgc
     1381 agcagcagga ttagttgcag catcaattgt taattgtgga gcagcaagag cagcacaaaa
     1441 tgttgcatca acaattcttt actacaacga tattcttgaa tatgaaacag gattaccagg
     1501 aactgattac ggaagatcag aaggaactgc agttggattt tcatttttct cacactcaat
     1561 ttatggaggc ggaggaccag gaatttttac aggaaatcac gttgttacta gacactcaaa
     1621 aggatttgca attccaccag ttacagcagc attatgtgtt gatgcaggaa cacagatgtt
     1681 ttcaccagaa aaaacttcag cattagttgg agcagtttac tcagatattc cagaatttag
     1741 agaaccactt aaacacgtta ttgaaggagc aattgaaatt aaagataaaa tttaaattat
     1801 caattgctca tatatctttt attcccggta tttaacttat tggtaggtaa tattatgatt
     1861 gaaattgaag tttttccaca cagattttta aaagcagata caactgaaaa atttcttaac
     1921 gatatttact caattgaaac tgttgaaaga gttattgttc acggacaacc attaccaaaa
     1981 actgtttatt acggaccagc aagaggaaca ccagttaatc acccagaaag aaaagaagtt
     2041 cttgttaacg gagttccagt tgaacttaca gttatggcag gaagattttg gattcttctt
     2101 gaaaacgata acgaacttaa caaaattgaa gaaatttgta aatcattatt tccatatgga
     2161 tacaatttaa atgttggaaa atttactaga gattcaccaa caactacaga ttacatgaaa
     2221 tacggagaaa aacttgttaa ctcaatggat agaaaaatga ttggacttac agatccaaga
     2281 tcaaaatttg catcatcagt taaaatgatt ccaaaaaacg aaaacgcaga atgtgaacag
     2341 aattaagagg atagtgattt tatgccagtt ggaagaaaag aacaaattgt tgattgtaga
     2401 gcagttagag gattaggaga aggtggagga ttagcacaga gaggaacatt tgcagaagga
     2461 cttaaaaacg atgttgttgt tgttgcaatg tcaccaggaa gaagacacat tacaaaacca
     2521 gtttgtgaaa ttacttacgg aattagagaa gcaggaattc aaacttcagt tcttgttctt
     2581 aacgcaggat caggattacc acacgataca ccaaaaggat cattaggatc aacttttgga
     2641 ttaaaaccag aagaagcaaa acaggttaac agacacaaac tttgtattgt tcaccttgga
     2701 aacgttaaat cacacattat ttacaaagca agattatttc ttagatacgt taaaattcca
     2761 acaattattg tttgtcaaac tccaattgat atggaagatt ttgcaaaaat tggagttaaa
     2821 acaaaaaacg ttatgccagt taatccagat acagaaggaa ctattgttga tattattact
     2881 ggagttgtta gaggagaatc agcaacacag cttaaaattg atgaaactat tgaaaaaatt
     2941 aaaaatcact taaattaaat taaaaggtga ttctatggat tacaaagatg atgatgataa
     3001 aggcgccagc tggtcacacc cacaatttga aaaaggaggt ggatcaggag gtggatcagg
     3061 aggttcagca tggtcacatc ctcagtttga aaaatcagca atggcatata ctccacaatt
     3121 ttgtccagga tcaacaactg ttgcacagaa tagaagagat cacatgaatc cagatgttca
     3181 acttgaaaaa cttagagaaa ttccagatga tgaaattgtt aaagttatgg gacacagaca
     3241 gccaggagaa gattacaaaa cagttcaccc accattagaa gaaatggata tgccagaaga
     3301 ttttgttaga gatattgttg aaccaatgtc aggagcaaaa gaaggacaca gaattagata
     3361 cattcaattt gcagattcaa tgtacaatgc accagcacag ccatatgata gagcaagaac
     3421 ttacatgtgg agatttagag gagttgatac aggaactctt tcaggaagac aagttattga
     3481 aatgagagaa tcaaaccttg aagaaatgtc aaaaaacttt tgtattgata cagcattttt
     3541 cgatccagca acttgtggaa tgagaggagc aacagttcac ggacactcac ttagacttga
     3601 tgaaaacgga cttatgtttg atgcattaca aagatatgtt tttgatgaaa aaacaggaca
     3661 cgttctttac gttaaagatc aggttggaag accattagat gaaccagttg attttggaga
     3721 accattatca actgaaaaac ttaaagaaat tacaactatt tacagaaaag attcaattgc
     3781 aatgagagat gatgaagaaa ttgttacagt tgttaaaaga attcacagag caagaactct
     3841 tggaggatat atgccaacag aaaaaatttt tgaaggatta taagtgaaat acgatacagc
     3901 aaatattgaa tatattactt aacctacctc tacttacctt ttgaggtgaa ttttatggaa
     3961 gcagaaaaga aattatttct taaagcatta aaagaaaaat ttgatgaaga tccaaaagaa
     4021 aaatacacaa aattttacac ttacggagga tggaaacaat caaaaagaaa aactgaattt
     4081 gttgaagcag caaaagatat tgcagataaa agaggtggaa tgccaggata caatccagat
     4141 attggagttc cattaggaca gagaaaaatg atgtcataca aactttcagg aacagatgat
     4201 tacgttgaag gagatgattt acactttatg aacaacgcag caattcaaca gcttcacgat
     4261 gatattagaa gaactgttat tgttggaatg gatacaggac acatggttct tgaaaaacga
     4321 ttaggagttg aagttacacc agaaactatt aacgaataca tggaaactat taatcacgca
     4381 ttacctggag gagcagttgt tcaagaacac atggttgaag ttgatccagc attagcatgg
     4441 gattcatacg caaaaatttt tacaggagat gatgaacttg cagatgaatt agataaaaga
     4501 gttttagttg atattaacaa attatttcca gaagaacaag cagaagaact taaagcagca
     4561 attggaaaga aaacatacca gatttcaaga gttccaacat tagttggaag agtttgtgat
     4621 ggaggaacta ttgcaagatg gtcagcaatg cagattggaa tgtcatttat tacagcatac
     4681 aaactttgtg caggagaagc agcaattgca gatttttcat acgcagcaaa acacgcagat
     4741 gttattggaa tgggaactgc attaccagca agaagatcaa gaggagcaaa tgaacctgga
     4801 ggaattccat ttggaattct ttgtgatgtt gttcaaacaa ctagaatttc agatgatcca
     4861 gttgaacagt cattagaagt tgttgcaaca ggagcaatgt tatatgatca aatttggtta
     4921 ggagcataca tgtctggagg agttggattt acacagtatg caactgcagc atacacagat
     4981 gatattcttg atgatttttg ttactacgga cttgattacg ttgaaaagaa atttggaaga
     5041 tgtggaacaa aagcaactat ggatgtagtt gaagatattg caacagaagt tactctttac
     5101 tcacttgaac aatacgatga atacccaact ttattagaag atcactttgg aggatcacag
     5161 agagcagcag ttgcagcagc agcagcagga gtttcagttt gtatggcaac aggaaattca
     5221 aatgcaggag ttaacggatg gtacctttca caaattatgc acaaagaata ccactcaaga
     5281 cttggatttt acggatacga tttacaagat cagtgtggag catcaaactc actttcaatt
     5341 agaaacgatg aatcatcacc attagaactt agaggaccaa actacccaaa ttacgcaatg
     5401 aatgttggac accaaggaga atacgcagga attactcagg cagcacactc agcaagagga
     5461 gatgcatttg caatgaatcc attaattaaa gttgcatttg cagatccatc acttattttt
     5521 gattttgcaa gaccaagaaa atgttgtgca agaggagcat taagagaatt tgaagttgca
     5581 ggagaaagag atccaatttt accagcaaaa taaactagta gcggccgctg caggatatag
     5641 caaaagtggg acttaagttc ccacttttgc tattttataa aaaacgccct attcgtaata
     5701 tactattcaa aactaaatat tcattgaaat atttgggtaa taattaatat taatctaata
     5761 tttagatatt gagatataat aataacattt ttttaaaaaa tcatctattt tcatcttaat
     5821 tttatatatt aaagcccatt ttttgagtat tcaaattcaa attattgtgt tattaacatc
     5881 ttatatataa acttttctat ttaatgttaa tgaaaaagtg aatatatata catagagtaa
     5941 tgttatgatg tatatatcaa aaaaaactag aggagacgac cttccatgac agaatataaa
     6001 ccaacagtta gattagcaac aagagatgac gtacctaggg cagtaagaac tcttgcagca
     6061 gcttttgcag attatccagc aacaagacac acagttgatc cagatagaca cattgaaaga
     6121 gttacagaat tacaagaatt atttttaaca agagttggtc ttgatatagg taaagtatgg
     6181 gttgcagatg atggagcagc agttgcagtt tggacaacac cagaatcagt tgaagctggt
     6241 gctgtttttg cagaaattgg accaagaatg gctgaattat caggatcaag attagcagct
     6301 cagcaacaga tggaaggatt acttgcacca cacagaccaa aagaaccagc atggttttta
     6361 gcaacagttg gagtttcacc agatcaccag ggaaaaggtt taggttctgc tgttgtactt
     6421 cctggtgttg aagcagctga aagagcagga gttccagcat ttttagaaac atcagcacca
     6481 agaaatcttc cattttatga aagacttgga tttacagtta cagcagatgt tgaagttcca
     6541 gaaggaccaa gaacatggtg tatgacaaga aaaccaggag cataaccatg gctccgaccg
     6601 aagccacccg gggcggcccc gccgaccccg cacccgcccc cgaggcccac cgcgggggac
     6661 acaccgaaca cgccgaccct gctgaacacg cggcgcagtt cggtgcccag gagcggatcg
     6721 ggaattaatt cgaagctgct ggtgaaagag accctatctt acctgctaaa atctaagtta
     6781 attactaatt tattattaat ttattattag attgggtaaa atagtaaaag aaaactaaag
     6841 gaaacctaat atggtttcct ttttttatat atttttaatt cactgggggg atttaaaact
     6901 tcatttttaa tttaaaagga tctaggtgaa gatccttggt gcactctcag tacaatctgc
     6961 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     7021 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     7081 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     7141 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     7201 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7261 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7321 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7381 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7441 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7501 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7561 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7621 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7681 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7741 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7801 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7861 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7921 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7981 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     8041 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8101 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     8161 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     8221 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8281 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8341 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8401 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8461 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     8521 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8581 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8641 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8701 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8761 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8821 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8881 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8941 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     9001 gtacccatta tttagttttt ttttatcacc acaacgtata aacccccttt cagaagaaaa
     9061 cagagaatga aaacagattt taatttatat atcgtatcta taatttagtt tttttttatt
     9121 gcagttttag aagtgcgttt gtcctggagc gaaatacacg atatcgtgta gtctccttca
     9181 gatgaaaata atatttttaa ttttcagaaa tattactgga aaaatacctg aaaaatattt
     9241 gaaaaaaaaa catacaccca gcggggaaaa atcgaacatt ttgaaaaagt ctatttatag
     9301 ggtttaaaca gggccaagac ctgcgggata taccccaaaa atgttcgata aaaaaatcga
     9361 acatttttcg aacatttttt ataatgttcg atgtgtgcga gtatgttcga taaaaaaatc
     9421 gaacattttg aaagacaatt tgaagagata taaaaaaaat aatctattat aatatagata
     9481 tataatatgt tcgatatgtt cgatattttt tt
//
