LOCUS       sequence-379174-        8900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged MCRaeoBDGA in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8900)
  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 8900)
  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 mcrBDGA
FEATURES             Location/Qualifiers
     source          1..8900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2096..2119
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2129..2152,2153..2188,2189..2212)
                     /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(6069..6088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6188..6210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6248..6266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6334..6438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6439..6507,6508..7299)
                     /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(6657..6676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7470..8058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7959..7978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggtaatat ccttccttct ttcctgcata cttcaaaaat ccaaggtttt ttaagttatc
       61 gagataagaa ttactagatg aattcgcgcc cggttctaga tttattcata aaataggaac
      121 taaaatacca aaatagaaac atttatatac atttgatgtg tagggtatac cataaacctg
      181 ggaggtgtct catatggtta aatacgaaga taaaattaac ctttacgatg caaaaggaaa
      241 ccttgcagaa gaaaacgttc cacttgaagc aatttcacca ttatacaatc cagttgttaa
      301 agatatggtt aaatcaatta aaagaacagt tgcagttaac cttgcaggaa ttgaagcaac
      361 acttaaagca ggaactattg gaggaaaagc atgtagagtt ccaggaagaa ctcttgattt
      421 accaattgtt gatgaagcag aagcaattat ggataaagtt gaaaaaattc ttaaaatttc
      481 agaagatgat gatacaaata ttagaactat taatggagga aaacaagcag ttgttcaggt
      541 tccatcaaaa agacttgata ttgcagcaga atactcagtt tcaatgttat caacaggaat
      601 ggcacttaaa gaagcaatta ttgatacatt tgatgttgat atgtttgatg gatcaactgt
      661 tcacgcagca attgttggaa gatatccaca agttatggat tacatgggag gaaatattgc
      721 atcattatta ggagcaccat caaatatgga aggattagga tacgcactta gaaacattat
      781 ggttaaccac tacgttgcaa caacaaagaa aaatcttatg aacgcagttg cattttcatc
      841 aattatggaa caaactgcaa tgtttgaaat gggagatgca attggatcat ttgaaagaat
      901 gcacttatta ggacttggat accagggact taacgcagat aacattgtta ttgatttagt
      961 taaagcaaat gcatcaggaa cagttggaac tgttgttgca tcagttgttg aaagagcact
     1021 tgaagataaa gttattgcag aagataaaac aatgaactca ggatttacta tgtacaaacc
     1081 agttgatgtt gcaaaatgga atgcatacgc agcagcagga ttagttgcag catcaattgt
     1141 taattgtgga gcagcaagag cagcacaaaa tgttgcatca acaattcttt actacaacga
     1201 tattcttgaa tatgaaacag gattaccagg aactgattac ggaagatcag aaggaactgc
     1261 agttggattt tcatttttct cacactcaat ttatggaggc ggaggaccag gaatttttac
     1321 aggaaatcac gttgttacta gacactcaaa aggatttgca attccaccag ttacagcagc
     1381 attatgtgtt gatgcaggaa cacagatgtt ttcaccagaa aaaacttcag cattagttgg
     1441 agcagtttac tcagatattc cagaatttag agaaccactt aaacacgtta ttgaaggagc
     1501 aattgaaatt aaagataaaa tttaaattat caattgctca tatatctttt attcccggta
     1561 tttaacttat tggtaggtaa tattatgatt gaaattgaag tttttccaca cagattttta
     1621 aaagcagata caactgaaaa atttcttaac gatatttact caattgaaac tgttgaaaga
     1681 gttattgttc acggacaacc attaccaaaa actgtttatt acggaccagc aagaggaaca
     1741 ccagttaatc acccagaaag aaaagaagtt cttgttaacg gagttccagt tgaacttaca
     1801 gttatggcag gaagattttg gattcttctt gaaaacgata acgaacttaa caaaattgaa
     1861 gaaatttgta aatcattatt tccatatgga tacaatttaa atgttggaaa atttactaga
     1921 gattcaccaa caactacaga ttacatgaaa tacggagaaa aacttgttaa ctcaatggat
     1981 agaaaaatga ttggacttac agatccaaga tcaaaatttg catcatcagt taaaatgatt
     2041 ccaaaaaacg aaaacgcaga atgtgaacag aattaaatta aaaggtgatt ctatggatta
     2101 caaagatgat gatgataaag gcgccagctg gtcacaccca caatttgaaa aaggaggtgg
     2161 atcaggaggt ggatcaggag gttcagcatg gtcacatcct cagtttgaaa aatcagcaat
     2221 ggcatatact ccacaatttt gtccaggatc aacaactgtt gcacagaata gaagagatca
     2281 catgaatcca gatgttcaac ttgaaaaact tagagaaatt ccagatgatg aaattgttaa
     2341 agttatggga cacagacagc caggagaaga ttacaaaaca gttcacccac cattagaaga
     2401 aatggatatg ccagaagatt ttgttagaga tattgttgaa ccaatgtcag gagcaaaaga
     2461 aggacacaga attagataca ttcaatttgc agattcaatg tacaatgcac cagcacagcc
     2521 atatgataga gcaagaactt acatgtggag atttagagga gttgatacag gaactctttc
     2581 aggaagacaa gttattgaaa tgagagaatc aaaccttgaa gaaatgtcaa aaaacttttg
     2641 tattgataca gcatttttcg atccagcaac ttgtggaatg agaggagcaa cagttcacgg
     2701 acactcactt agacttgatg aaaacggact tatgtttgat gcattacaaa gatatgtttt
     2761 tgatgaaaaa acaggacacg ttctttacgt taaagatcag gttggaagac cattagatga
     2821 accagttgat tttggagaac cattatcaac tgaaaaactt aaagaaatta caactattta
     2881 cagaaaagat tcaattgcaa tgagagatga tgaagaaatt gttacagttg ttaaaagaat
     2941 tcacagagca agaactcttg gaggatatat gccaacagaa aaaatttttg aaggattata
     3001 agtgaaatac gatacagcaa atattgaata tattacttaa cctacctcta cttacctttt
     3061 gaggtgaatt ttatggaagc agaaaagaaa ttatttctta aagcattaaa agaaaaattt
     3121 gatgaagatc caaaagaaaa atacacaaaa ttttacactt acggaggatg gaaacaatca
     3181 aaaagaaaaa ctgaatttgt tgaagcagca aaagatattg cagataaaag aggtggaatg
     3241 ccaggataca atccagatat tggagttcca ttaggacaga gaaaaatgat gtcatacaaa
     3301 ctttcaggaa cagatgatta cgttgaagga gatgatttac actttatgaa caacgcagca
     3361 attcaacagc ttcacgatga tattagaaga actgttattg ttggaatgga tacaggacac
     3421 atggttcttg aaaaacgatt aggagttgaa gttacaccag aaactattaa cgaatacatg
     3481 gaaactatta atcacgcatt acctggagga gcagttgttc aagaacacat ggttgaagtt
     3541 gatccagcat tagcatggga ttcatacgca aaaattttta caggagatga tgaacttgca
     3601 gatgaattag ataaaagagt tttagttgat attaacaaat tatttccaga agaacaagca
     3661 gaagaactta aagcagcaat tggaaagaaa acataccaga tttcaagagt tccaacatta
     3721 gttggaagag tttgtgatgg aggaactatt gcaagatggt cagcaatgca gattggaatg
     3781 tcatttatta cagcatacaa actttgtgca ggagaagcag caattgcaga tttttcatac
     3841 gcagcaaaac acgcagatgt tattggaatg ggaactgcat taccagcaag aagatcaaga
     3901 ggagcaaatg aacctggagg aattccattt ggaattcttt gtgatgttgt tcaaacaact
     3961 agaatttcag atgatccagt tgaacagtca ttagaagttg ttgcaacagg agcaatgtta
     4021 tatgatcaaa tttggttagg agcatacatg tctggaggag ttggatttac acagtatgca
     4081 actgcagcat acacagatga tattcttgat gatttttgtt actacggact tgattacgtt
     4141 gaaaagaaat ttggaagatg tggaacaaaa gcaactatgg atgtagttga agatattgca
     4201 acagaagtta ctctttactc acttgaacaa tacgatgaat acccaacttt attagaagat
     4261 cactttggag gatcacagag agcagcagtt gcagcagcag cagcaggagt ttcagtttgt
     4321 atggcaacag gaaattcaaa tgcaggagtt aacggatggt acctttcaca aattatgcac
     4381 aaagaatacc actcaagact tggattttac ggatacgatt tacaagatca gtgtggagca
     4441 tcaaactcac tttcaattag aaacgatgaa tcatcaccat tagaacttag aggaccaaac
     4501 tacccaaatt acgcaatgaa tgttggacac caaggagaat acgcaggaat tactcaggca
     4561 gcacactcag caagaggaga tgcatttgca atgaatccat taattaaagt tgcatttgca
     4621 gatccatcac ttatttttga ttttgcaaga ccaagaaaat gttgtgcaag aggagcatta
     4681 agagaatttg aagttgcagg agaaagagat ccaattttac cagcaaaata aactagtagc
     4741 ggccgctgca ggatatagca aaagtgggac ttaagttccc acttttgcta ttttataaaa
     4801 aacgccctat tcgtaatata ctattcaaaa ctaaatattc attgaaatat ttgggtaata
     4861 attaatatta atctaatatt tagatattga gatataataa taacattttt ttaaaaaatc
     4921 atctattttc atcttaattt tatatattaa agcccatttt ttgagtattc aaattcaaat
     4981 tattgtgtta ttaacatctt atatataaac ttttctattt aatgttaatg aaaaagtgaa
     5041 tatatataca tagagtaatg ttatgatgta tatatcaaaa aaaactagag gagacgacct
     5101 tccatgacag aatataaacc aacagttaga ttagcaacaa gagatgacgt acctagggca
     5161 gtaagaactc ttgcagcagc ttttgcagat tatccagcaa caagacacac agttgatcca
     5221 gatagacaca ttgaaagagt tacagaatta caagaattat ttttaacaag agttggtctt
     5281 gatataggta aagtatgggt tgcagatgat ggagcagcag ttgcagtttg gacaacacca
     5341 gaatcagttg aagctggtgc tgtttttgca gaaattggac caagaatggc tgaattatca
     5401 ggatcaagat tagcagctca gcaacagatg gaaggattac ttgcaccaca cagaccaaaa
     5461 gaaccagcat ggtttttagc aacagttgga gtttcaccag atcaccaggg aaaaggttta
     5521 ggttctgctg ttgtacttcc tggtgttgaa gcagctgaaa gagcaggagt tccagcattt
     5581 ttagaaacat cagcaccaag aaatcttcca ttttatgaaa gacttggatt tacagttaca
     5641 gcagatgttg aagttccaga aggaccaaga acatggtgta tgacaagaaa accaggagca
     5701 taaccatggc tccgaccgaa gccacccggg gcggccccgc cgaccccgca cccgcccccg
     5761 aggcccaccg cgggggacac accgaacacg ccgaccctgc tgaacacgcg gcgcagttcg
     5821 gtgcccagga gcggatcggg aattaattcg aagctgctgg tgaaagagac cctatcttac
     5881 ctgctaaaat ctaagttaat tactaattta ttattaattt attattagat tgggtaaaat
     5941 agtaaaagaa aactaaagga aacctaatat ggtttccttt ttttatatat ttttaattca
     6001 ctggggggat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tccttggtgc
     6061 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6121 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6181 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6241 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6301 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6361 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6421 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6481 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6541 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6601 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6661 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6721 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6781 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6841 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6901 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6961 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7021 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7081 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7141 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7201 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7261 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7441 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7501 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7561 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7621 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7681 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7741 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7801 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7861 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7921 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7981 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8041 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8101 tggccttttg ctcacatggt acccattatt tagttttttt ttatcaccac aacgtataaa
     8161 ccccctttca gaagaaaaca gagaatgaaa acagatttta atttatatat cgtatctata
     8221 atttagtttt tttttattgc agttttagaa gtgcgtttgt cctggagcga aatacacgat
     8281 atcgtgtagt ctccttcaga tgaaaataat atttttaatt ttcagaaata ttactggaaa
     8341 aatacctgaa aaatatttga aaaaaaaaca tacacccagc ggggaaaaat cgaacatttt
     8401 gaaaaagtct atttataggg tttaaacagg gccaagacct gcgggatata ccccaaaaat
     8461 gttcgataaa aaaatcgaac atttttcgaa cattttttat aatgttcgat gtgtgcgagt
     8521 atgttcgata aaaaaatcga acattttgaa agacaatttg aagagatata aaaaaaataa
     8581 tctattataa tatagatata taatatgttc gatatgttcg atattttttt atatattata
     8641 ttatatttaa ccttgtattg attattgaat aatatttatt attacttttt ttgatttctt
     8701 gtaaaaaatc gaacatattt tacatattgt aaaaaataaa aaagatttaa aaaaagtgtc
     8761 cgaaaaagtt aaacaaacat tattattaaa atgttagttt aatatgtatc ttgatatgct
     8821 cttctttctt tcatgcttaa acttgtataa ctgtatgctt ttttcatttc ctttattttg
     8881 tcctttcctt tctttgttat
//
