LOCUS       sequence-379059-        9503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CA-tagged MCRaeo in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9503)
  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 9503)
  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-CA
FEATURES             Location/Qualifiers
     source          1..9503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5199..5222
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(5232..5255,5256..5291,5292..5315)
                     /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(6656..6675)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6775..6797
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6835..6853)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6921..7025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7026..7094,7095..7886)
                     /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(7244..7263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8057..8645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8546..8565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg catacttcaa aaatccaagg ttttttaagt tatcgagata agaattacta
       61 gatgaattcg cgcccggttc tagatttatt cataaaatag gaactaaaat accaaaatag
      121 aaacatttat atacatttga tgtgtagggt ataccataaa cctgggaggt gtctcatatg
      181 gttaaatacg aagataaaat taacctttac gatgcaaaag gaaaccttgc agaagaaaac
      241 gttccacttg aagcaatttc accattatac aatccagttg ttaaagatat ggttaaatca
      301 attaaaagaa cagttgcagt taaccttgca ggaattgaag caacacttaa agcaggaact
      361 attggaggaa aagcatgtag agttccagga agaactcttg atttaccaat tgttgatgaa
      421 gcagaagcaa ttatggataa agttgaaaaa attcttaaaa tttcagaaga tgatgataca
      481 aatattagaa ctattaatgg aggaaaacaa gcagttgttc aggttccatc aaaaagactt
      541 gatattgcag cagaatactc agtttcaatg ttatcaacag gaatggcact taaagaagca
      601 attattgata catttgatgt tgatatgttt gatggatcaa ctgttcacgc agcaattgtt
      661 ggaagatatc cacaagttat ggattacatg ggaggaaata ttgcatcatt attaggagca
      721 ccatcaaata tggaaggatt aggatacgca cttagaaaca ttatggttaa ccactacgtt
      781 gcaacaacaa agaaaaatct tatgaacgca gttgcatttt catcaattat ggaacaaact
      841 gcaatgtttg aaatgggaga tgcaattgga tcatttgaaa gaatgcactt attaggactt
      901 ggataccagg gacttaacgc agataacatt gttattgatt tagttaaagc aaatgcatca
      961 ggaacagttg gaactgttgt tgcatcagtt gttgaaagag cacttgaaga taaagttatt
     1021 gcagaagata aaacaatgaa ctcaggattt actatgtaca aaccagttga tgttgcaaaa
     1081 tggaatgcat acgcagcagc aggattagtt gcagcatcaa ttgttaattg tggagcagca
     1141 agagcagcac aaaatgttgc atcaacaatt ctttactaca acgatattct tgaatatgaa
     1201 acaggattac caggaactga ttacggaaga tcagaaggaa ctgcagttgg attttcattt
     1261 ttctcacact caatttatgg aggcggagga ccaggaattt ttacaggaaa tcacgttgtt
     1321 actagacact caaaaggatt tgcaattcca ccagttacag cagcattatg tgttgatgca
     1381 ggaacacaga tgttttcacc agaaaaaact tcagcattag ttggagcagt ttactcagat
     1441 attccagaat ttagagaacc acttaaacac gttattgaag gagcaattga aattaaagat
     1501 aaaatttaaa ttatcaattg ctcatatatc ttttattccc ggtatttaac ttattggtag
     1561 gtaatattat gattgaaatt gaagtttttc cacacagatt tttaaaagca gatacaactg
     1621 aaaaatttct taacgatatt tactcaattg aaactgttga aagagttatt gttcacggac
     1681 aaccattacc aaaaactgtt tattacggac cagcaagagg aacaccagtt aatcacccag
     1741 aaagaaaaga agttcttgtt aacggagttc cagttgaact tacagttatg gcaggaagat
     1801 tttggattct tcttgaaaac gataacgaac ttaacaaaat tgaagaaatt tgtaaatcat
     1861 tatttccata tggatacaat ttaaatgttg gaaaatttac tagagattca ccaacaacta
     1921 cagattacat gaaatacgga gaaaaacttg ttaactcaat ggatagaaaa atgattggac
     1981 ttacagatcc aagatcaaaa tttgcatcat cagttaaaat gattccaaaa aacgaaaacg
     2041 cagaatgtga acagaattaa gaggatagtg attttatgcc agttggaaga aaagaacaaa
     2101 ttgttgattg tagagcagtt agaggattag gagaaggtgg aggattagca cagagaggaa
     2161 catttgcaga aggacttaaa aacgatgttg ttgttgttgc aatgtcacca ggaagaagac
     2221 acattacaaa accagtttgt gaaattactt acggaattag agaagcagga attcaaactt
     2281 cagttcttgt tcttaacgca ggatcaggat taccacacga tacaccaaaa ggatcattag
     2341 gatcaacttt tggattaaaa ccagaagaag caaaacaggt taacagacac aaactttgta
     2401 ttgttcacct tggaaacgtt aaatcacaca ttatttacaa agcaagatta tttcttagat
     2461 acgttaaaat tccaacaatt attgtttgtc aaactccaat tgatatggaa gattttgcaa
     2521 aaattggagt taaaacaaaa aacgttatgc cagttaatcc agatacagaa ggaactattg
     2581 ttgatattat tactggagtt gttagaggag aatcagcaac acagcttaaa attgatgaaa
     2641 ctattgaaaa aattaaaaat cacttaaatt aaattaaaag gtgattctat ggcatatact
     2701 ccacaatttt gtccaggatc aacaactgtt gcacagaata gaagagatca catgaatcca
     2761 gatgttcaac ttgaaaaact tagagaaatt ccagatgatg aaattgttaa agttatggga
     2821 cacagacagc caggagaaga ttacaaaaca gttcacccac cattagaaga aatggatatg
     2881 ccagaagatt ttgttagaga tattgttgaa ccaatgtcag gagcaaaaga aggacacaga
     2941 attagataca ttcaatttgc agattcaatg tacaatgcac cagcacagcc atatgataga
     3001 gcaagaactt acatgtggag atttagagga gttgatacag gaactctttc aggaagacaa
     3061 gttattgaaa tgagagaatc aaaccttgaa gaaatgtcaa aaaacttttg tattgataca
     3121 gcatttttcg atccagcaac ttgtggaatg agaggagcaa cagttcacgg acactcactt
     3181 agacttgatg aaaacggact tatgtttgat gcattacaaa gatatgtttt tgatgaaaaa
     3241 acaggacacg ttctttacgt taaagatcag gttggaagac cattagatga accagttgat
     3301 tttggagaac cattatcaac tgaaaaactt aaagaaatta caactattta cagaaaagat
     3361 tcaattgcaa tgagagatga tgaagaaatt gttacagttg ttaaaagaat tcacagagca
     3421 agaactcttg gaggatatat gccaacagaa aaaatttttg aaggattata agtgaaatac
     3481 gatacagcaa atattgaata tattacttaa cctacctcta cttacctttt gaggtgaatt
     3541 ttatggaagc agaaaagaaa ttatttctta aagcattaaa agaaaaattt gatgaagatc
     3601 caaaagaaaa atacacaaaa ttttacactt acggaggatg gaaacaatca aaaagaaaaa
     3661 ctgaatttgt tgaagcagca aaagatattg cagataaaag aggtggaatg ccaggataca
     3721 atccagatat tggagttcca ttaggacaga gaaaaatgat gtcatacaaa ctttcaggaa
     3781 cagatgatta cgttgaagga gatgatttac actttatgaa caacgcagca attcaacagc
     3841 ttcacgatga tattagaaga actgttattg ttggaatgga tacaggacac atggttcttg
     3901 aaaaacgatt aggagttgaa gttacaccag aaactattaa cgaatacatg gaaactatta
     3961 atcacgcatt acctggagga gcagttgttc aagaacacat ggttgaagtt gatccagcat
     4021 tagcatggga ttcatacgca aaaattttta caggagatga tgaacttgca gatgaattag
     4081 ataaaagagt tttagttgat attaacaaat tatttccaga agaacaagca gaagaactta
     4141 aagcagcaat tggaaagaaa acataccaga tttcaagagt tccaacatta gttggaagag
     4201 tttgtgatgg aggaactatt gcaagatggt cagcaatgca gattggaatg tcatttatta
     4261 cagcatacaa actttgtgca ggagaagcag caattgcaga tttttcatac gcagcaaaac
     4321 acgcagatgt tattggaatg ggaactgcat taccagcaag aagatcaaga ggagcaaatg
     4381 aacctggagg aattccattt ggaattcttt gtgatgttgt tcaaacaact agaatttcag
     4441 atgatccagt tgaacagtca ttagaagttg ttgcaacagg agcaatgtta tatgatcaaa
     4501 tttggttagg agcatacatg tctggaggag ttggatttac acagtatgca actgcagcat
     4561 acacagatga tattcttgat gatttttgtt actacggact tgattacgtt gaaaagaaat
     4621 ttggaagatg tggaacaaaa gcaactatgg atgtagttga agatattgca acagaagtta
     4681 ctctttactc acttgaacaa tacgatgaat acccaacttt attagaagat cactttggag
     4741 gatcacagag agcagcagtt gcagcagcag cagcaggagt ttcagtttgt atggcaacag
     4801 gaaattcaaa tgcaggagtt aacggatggt acctttcaca aattatgcac aaagaatacc
     4861 actcaagact tggattttac ggatacgatt tacaagatca gtgtggagca tcaaactcac
     4921 tttcaattag aaacgatgaa tcatcaccat tagaacttag aggaccaaac tacccaaatt
     4981 acgcaatgaa tgttggacac caaggagaat acgcaggaat tactcaggca gcacactcag
     5041 caagaggaga tgcatttgca atgaatccat taattaaagt tgcatttgca gatccatcac
     5101 ttatttttga ttttgcaaga ccaagaaaat gttgtgcaag aggagcatta agagaatttg
     5161 aagttgcagg agaaagagat ccaattttac cagcaaaaga ttacaaagat gatgatgata
     5221 aaggcgccag ctggtcacac ccacaatttg aaaaaggagg tggatcagga ggtggatcag
     5281 gaggttcagc atggtcacat cctcagtttg aaaaataaac tagtagcggc cgctgcagga
     5341 tatagcaaaa gtgggactta agttcccact tttgctattt tataaaaaac gccctattcg
     5401 taatatacta ttcaaaacta aatattcatt gaaatatttg ggtaataatt aatattaatc
     5461 taatatttag atattgagat ataataataa cattttttta aaaaatcatc tattttcatc
     5521 ttaattttat atattaaagc ccattttttg agtattcaaa ttcaaattat tgtgttatta
     5581 acatcttata tataaacttt tctatttaat gttaatgaaa aagtgaatat atatacatag
     5641 agtaatgtta tgatgtatat atcaaaaaaa actagaggag acgaccttcc atgacagaat
     5701 ataaaccaac agttagatta gcaacaagag atgacgtacc tagggcagta agaactcttg
     5761 cagcagcttt tgcagattat ccagcaacaa gacacacagt tgatccagat agacacattg
     5821 aaagagttac agaattacaa gaattatttt taacaagagt tggtcttgat ataggtaaag
     5881 tatgggttgc agatgatgga gcagcagttg cagtttggac aacaccagaa tcagttgaag
     5941 ctggtgctgt ttttgcagaa attggaccaa gaatggctga attatcagga tcaagattag
     6001 cagctcagca acagatggaa ggattacttg caccacacag accaaaagaa ccagcatggt
     6061 ttttagcaac agttggagtt tcaccagatc accagggaaa aggtttaggt tctgctgttg
     6121 tacttcctgg tgttgaagca gctgaaagag caggagttcc agcattttta gaaacatcag
     6181 caccaagaaa tcttccattt tatgaaagac ttggatttac agttacagca gatgttgaag
     6241 ttccagaagg accaagaaca tggtgtatga caagaaaacc aggagcataa ccatggctcc
     6301 gaccgaagcc acccggggcg gccccgccga ccccgcaccc gcccccgagg cccaccgcgg
     6361 gggacacacc gaacacgccg accctgctga acacgcggcg cagttcggtg cccaggagcg
     6421 gatcgggaat taattcgaag ctgctggtga aagagaccct atcttacctg ctaaaatcta
     6481 agttaattac taatttatta ttaatttatt attagattgg gtaaaatagt aaaagaaaac
     6541 taaaggaaac ctaatatggt ttcctttttt tatatatttt taattcactg gggggattta
     6601 aaacttcatt tttaatttaa aaggatctag gtgaagatcc ttggtgcact ctcagtacaa
     6661 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6721 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6781 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     6841 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7081 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7141 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7201 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7261 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7321 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7381 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7441 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7501 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7561 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7621 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7681 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7741 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7801 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7861 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8701 acatggtacc cattatttag ttttttttta tcaccacaac gtataaaccc cctttcagaa
     8761 gaaaacagag aatgaaaaca gattttaatt tatatatcgt atctataatt tagttttttt
     8821 ttattgcagt tttagaagtg cgtttgtcct ggagcgaaat acacgatatc gtgtagtctc
     8881 cttcagatga aaataatatt tttaattttc agaaatatta ctggaaaaat acctgaaaaa
     8941 tatttgaaaa aaaaacatac acccagcggg gaaaaatcga acattttgaa aaagtctatt
     9001 tatagggttt aaacagggcc aagacctgcg ggatataccc caaaaatgtt cgataaaaaa
     9061 atcgaacatt tttcgaacat tttttataat gttcgatgtg tgcgagtatg ttcgataaaa
     9121 aaatcgaaca ttttgaaaga caatttgaag agatataaaa aaaataatct attataatat
     9181 agatatataa tatgttcgat atgttcgata tttttttata tattatatta tatttaacct
     9241 tgtattgatt attgaataat atttattatt actttttttg atttcttgta aaaaatcgaa
     9301 catattttac atattgtaaa aaataaaaaa gatttaaaaa aagtgtccga aaaagttaaa
     9361 caaacattat tattaaaatg ttagtttaat atgtatcttg atatgctctt ctttctttca
     9421 tgcttaaact tgtataactg tatgcttttt tcatttcctt tattttgtcc tttcctttct
     9481 ttgttatctg gtaatatcct tcc
//
