LOCUS       sequence-379180-        8399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NG-tagged ANME1G37 MCR in Methanococcus maripaludis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8399)
  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 8399)
  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-ANME1G37 MCR-NG
FEATURES             Location/Qualifiers
     source          1..8399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1578..1601
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1611..1634,1635..1670,1671..1694)
                     /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"
     CDS             2067..2078
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5604..5623)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5723..5745
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5783..5801)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5869..5973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5974..6042,6043..6834)
                     /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(6192..6211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7005..7593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7494..7513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catttccttt attttgtcct ttcctttctt tgttatctgg taatatcctt ccttctttcc
       61 tgcatacttc aaaaatccaa ggttttttaa gttatcgaga taagaattac tagatgaatt
      121 cgcgcccggt tctagattta ttcataaaat aggaactaaa ataccaaaat agaaacattt
      181 atatacattt gatgtgtagg gtataccata aacctgggag gtgtctcata tggcagatga
      241 agttgattta tacgatgata gaggaaacca attagcatca aacgttccat tagaagcaat
      301 ttcaccatta aaaaatccag caattaagaa aattgttaac cttacaatta gaactggagc
      361 agttgattta gcaaaacttg aaaacaaatt agcaacagga actattgcag gaaaaggaat
      421 ggttattaga ggagttggac acgattttcc aattttagca aatgcagaag caatagcaga
      481 acaagttgaa gatatgctta gagtttcaga agatgatgat acaaacgttg aacttatggc
      541 aggtggaaaa agaatgttaa ctcaggttcc atcagcaaga attctttcag attactcagt
      601 tggattaaca gcagcaatgg gagcattaac tcacgcaatt attgatgttt gtgatgtttc
      661 aatgtgggaa gcaccatatg ttcacgcagc agtttgggga atgtacccac aggatccaga
      721 tccagcagat ggagcagtta aaatgttagt tgatgttcca attaaaaacg aaggaccagg
      781 atttgttctt agaaacattc cagttaacca cttagcagca acagttagaa aaagagcaat
      841 gcaaggagca gcattagcaa tgattttaga agaagcagca cagtttgaaa tgggaaattg
      901 tatgggacca cacgaaagag gacacttatt agatttagca tatgaaggat taaatgcaaa
      961 caacatgctt agagatttaa ttaaagaaaa tggaaaaaca ggaactttag cagatgttgt
     1021 ttactcaact gttgaaagag caaaagcaga tggagttatt aaaccactta aagaaatgcc
     1081 atcaggattt gttgtttacg atgcagatga tatgcaatta tggaatgcat acgcatcagc
     1141 aggaatgtta gcagcagttt gtgttaattg tgcatcaatg agagcaggac agccagttcc
     1201 atcaaacatt atgtactcat gttgtcttat ggaaagagaa acaggattac caggaccaga
     1261 ttttggaatg gcacaaggag catcagtttc atcatcattt ttctcacact caatttatgg
     1321 aggcggagga ccaggagtat tttatggaaa ccacattgtt actagacacg caaaaggaca
     1381 gtttattcca tgtttttgtg cagcaatgtg tattgatgca gatacaatgt actttattcc
     1441 agcaagaact tcagcattat acggagaagt tttaggagca attccagaat ttgcagaacc
     1501 aatgaaagca gttgcagaag gagcacaaga aattatgtaa aaggagaaaa aaaggagaaa
     1561 ggaggaaaaa gaaaatggat tacaaagatg atgatgataa aggcgccagc tggtcacacc
     1621 cacaatttga aaaaggaggt ggatcaggag gtggatcagg aggttcagca tggtcacatc
     1681 ctcagtttga aaaatcagca atgccacaat atacagcagg atcatcacac attgcacaga
     1741 acagaagaaa ctacatggat ccaaactaca cttacgaaaa acttagagat attgcagaag
     1801 aagatattgt tagacttctt gcacacagag caccaggaga agaatataaa tcaattcacc
     1861 caccattaga agaaatggaa gaaccagaat gtgcagttag agaaatggtt gaaccaacag
     1921 aaggagcaaa agcaggagat agaattagat acgttcaata cacagattca atgatgtttg
     1981 caccaattac tccatatcag agagcatggg cagcatatac tagatacaaa ggagttgatc
     2041 caggagttct ttcaggaaga acaattattg aaggaagaga aagagatatt gaaaaaatta
     2101 ctaaagaaca aattgattgt gaactttacg atacagcaag aactggatta agaggaagaa
     2161 cagttcacgg acacgcagtt agattagatg aagatggaat gatgtttgat gcattaagaa
     2221 gatggtcaaa aggaaaaggt ggagaagtta tttacgttaa agatatgatt ggaggagcaa
     2281 tggataaaga agttgttctt ggaaaaccac ttccagaaga tgaacttctt aaaagaacaa
     2341 ctatgtacag aaatgcacag ggaggagttt ggcaggaagt tgatgatcca gaatcaatgg
     2401 atgttacagc acaaattcac tggaaaagaa ctgttggagg atttcagcca tggaagaaaa
     2461 tgtcagaaat taaaggtgga aagaaagata ttggagttaa agatttaaaa ttatttgtac
     2521 cgagaggagg tgtagaataa aatgccatac agagatgttc aacacaattt tcttaaagca
     2581 atgtcagata aatttgcaga aaaaccagaa tcaacaacta caaaatttta cgtttacgga
     2641 ggaattgcac agaaaggtgg aatgagaaaa agagaattta ttgaagaagc aaaaagaatg
     2701 gttgaaaaca gaactgttag aacaccagga tacaatccag atgttggaat gccacaagga
     2761 cagagatatt taatgccata catgatgaac cacactgata ttatggttaa cgcagatgat
     2821 ttacactgga ttaacaatgc agcaatgcaa cagtgttggg atgatatgaa aagaggaatt
     2881 attcttggat tagatgatgc acacggatta ttagaagcaa gacttggaaa agaagttact
     2941 ccagatacaa tttcacacta catggaagtt cttaatcacg cattacctgg aggagcagtt
     3001 attcaagaac acatggttga aactaaacca ttattagtta acgattcata cgcaaaaatc
     3061 tttacaggag atgatgattt agcagatgca ttagatagaa gatttgtttt agatattaac
     3121 aaagaatttc cagcaggatg ggaaaaacca ggagaacaag cagatcagtt aaaagatgca
     3181 ataggaaaga aaatttggca aattttatgg atgccaactg ttgttggaag aatgactgat
     3241 ggaggaacaa tgtttagatg ggttggaatg caggttggaa tgactatgat taacgcatac
     3301 aaaatgtgtg caggagaatc agcaacagga gaatttgcat attacgcaaa acacgcagca
     3361 gttgttaaac tttcaaactt tatgccagtt aaaagagcaa gatcacagaa tgaatgttca
     3421 ggaatgccat taggaattaa tgcagattca acaagatcac cagcattatt tccaaatgat
     3481 ccaattagag cagaattaga atcaatagca gttgcagcaa tggtttacga tcaattatgg
     3541 tttggaactt acatgtctgg aggagttgga tttacacagt atgcatcagc aacttacaca
     3601 gataacattt tagaagattt ttgttacaaa ggatgtgaaa ttggaagaga ttacattaac
     3661 gaagaaaata atggagaatt attaaaagga gataaactta acatggatat tttagaaaaa
     3721 attattagag cagaaaacga ttactgtctt actcaatacg aagcataccc aacagttgca
     3781 gaatcacact ttggaggatc agttagagca tgttgtgcag cagcaggatg tggatcagca
     3841 gttgcatgtg caactggatt agcacagcca acattatcag catggtcaat gtcaatgtta
     3901 ggacactacg aaagagttgg aagattagga ttttacggat acgatttaca agatcagtgt
     3961 actgcatgtg gatcatattc ataccaatca gatgaaggaa tgccatttga aatgagagga
     4021 gttaactacc caaactacgc aatgaacgtt ggacaccagt cagcatacgg aggattagtt
     4081 gcaggagcac acttagcaaa taaagatgca tgggttcttt caccattatg gaaagttgca
     4141 ttttcagata gagatttacc atttgataga ggatatgtta caagagaata cggaagagga
     4201 gcattaagag aatttaaacc agcaggagaa agagatttaa ttattggagg atattacgga
     4261 agataaacta gtagcggccg ctgcaggata tagcaaaagt gggacttaag ttcccacttt
     4321 tgctatttta taaaaaacgc cctattcgta atatactatt caaaactaaa tattcattga
     4381 aatatttggg taataattaa tattaatcta atatttagat attgagatat aataataaca
     4441 tttttttaaa aaatcatcta ttttcatctt aattttatat attaaagccc attttttgag
     4501 tattcaaatt caaattattg tgttattaac atcttatata taaacttttc tatttaatgt
     4561 taatgaaaaa gtgaatatat atacatagag taatgttatg atgtatatat caaaaaaaac
     4621 tagaggagac gaccttccat gacagaatat aaaccaacag ttagattagc aacaagagat
     4681 gacgtaccta gggcagtaag aactcttgca gcagcttttg cagattatcc agcaacaaga
     4741 cacacagttg atccagatag acacattgaa agagttacag aattacaaga attattttta
     4801 acaagagttg gtcttgatat aggtaaagta tgggttgcag atgatggagc agcagttgca
     4861 gtttggacaa caccagaatc agttgaagct ggtgctgttt ttgcagaaat tggaccaaga
     4921 atggctgaat tatcaggatc aagattagca gctcagcaac agatggaagg attacttgca
     4981 ccacacagac caaaagaacc agcatggttt ttagcaacag ttggagtttc accagatcac
     5041 cagggaaaag gtttaggttc tgctgttgta cttcctggtg ttgaagcagc tgaaagagca
     5101 ggagttccag catttttaga aacatcagca ccaagaaatc ttccatttta tgaaagactt
     5161 ggatttacag ttacagcaga tgttgaagtt ccagaaggac caagaacatg gtgtatgaca
     5221 agaaaaccag gagcataacc atggctccga ccgaagccac ccggggcggc cccgccgacc
     5281 ccgcacccgc ccccgaggcc caccgcgggg gacacaccga acacgccgac cctgctgaac
     5341 acgcggcgca gttcggtgcc caggagcgga tcgggaatta attcgaagct gctggtgaaa
     5401 gagaccctat cttacctgct aaaatctaag ttaattacta atttattatt aatttattat
     5461 tagattgggt aaaatagtaa aagaaaacta aaggaaacct aatatggttt ccttttttta
     5521 tatattttta attcactggg gggatttaaa acttcatttt taatttaaaa ggatctaggt
     5581 gaagatcctt ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5641 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5701 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5761 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5821 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5941 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6001 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6061 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6121 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6181 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6241 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6301 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6361 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6421 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6481 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6541 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6601 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6661 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6721 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6781 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6841 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7141 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7621 ttcctggcct tttgctggcc ttttgctcac atggtaccca ttatttagtt tttttttatc
     7681 accacaacgt ataaaccccc tttcagaaga aaacagagaa tgaaaacaga ttttaattta
     7741 tatatcgtat ctataattta gttttttttt attgcagttt tagaagtgcg tttgtcctgg
     7801 agcgaaatac acgatatcgt gtagtctcct tcagatgaaa ataatatttt taattttcag
     7861 aaatattact ggaaaaatac ctgaaaaata tttgaaaaaa aaacatacac ccagcgggga
     7921 aaaatcgaac attttgaaaa agtctattta tagggtttaa acagggccaa gacctgcggg
     7981 atatacccca aaaatgttcg ataaaaaaat cgaacatttt tcgaacattt tttataatgt
     8041 tcgatgtgtg cgagtatgtt cgataaaaaa atcgaacatt ttgaaagaca atttgaagag
     8101 atataaaaaa aataatctat tataatatag atatataata tgttcgatat gttcgatatt
     8161 tttttatata ttatattata tttaaccttg tattgattat tgaataatat ttattattac
     8221 tttttttgat ttcttgtaaa aaatcgaaca tattttacat attgtaaaaa ataaaaaaga
     8281 tttaaaaaaa gtgtccgaaa aagttaaaca aacattatta ttaaaatgtt agtttaatat
     8341 gtatcttgat atgctcttct ttctttcatg cttaaacttg tataactgta tgctttttt
//
