LOCUS       sequence-300482-        3314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses M.Ocy1ORF8430P DNA methyltransferase (M.SpeI ortholog) in 
            E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3314)
  AUTHORS   Matsumura I
  TITLE     Methylase-assisted subcloning for high throughput BioBrick assembly.
  JOURNAL   PeerJ. 2020 Sep 11;8:e9841. doi: 10.7717/peerj.9841. eCollection 
            2020.
  PUBMED    32974095
REFERENCE   2  (bases 1 to 3314)
  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: "PeerJ."; 
            date: "2020-09-11"; volume: "8:e9841. doi"; pages: " 
            10.7717/peerj.9841. eCollection 2020"
COMMENT     Sequence Label: Prham-M.Ocy1ORF8430P-p15A-aadA
FEATURES             Location/Qualifiers
     source          1..3314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        141..259
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     rep_origin      1285..1830
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1731..1750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1947..1964
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2098..2189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2190..2981
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(3152..3199)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3195..3213
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3251..3295)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
ORIGIN
        1 ctagaggatg tcgtcaacac ggcgaaatag taatcacgag gtcaggttct taccttaaat
       61 tttcgacgga aaaccacgta aaaaacgtcg atttttcaag atacagcgtg aattttcagg
      121 aaatgcggtg agcatcacat caccacaatt cagcaaattg tgaacatcat cacgttcatc
      181 tttccctggt tgccaatggc ccattttcct gtcagtaacg agaaggtcgc gaattgaggc
      241 gctttttaga ctggtcgtaa tgaaattcag ctactagagc atgcgtactt agaggaaata
      301 aaccatgaag aaagcgccgc tcaacaaagt tctgatcggt gactgccgtg aagttatgaa
      361 atccctgccg gaaaactacg tttctgcgtg catcaccgac ccaccgtaca actacgaatt
      421 tatcggtcac aaatggaacg actctgaaat caaacgtcgt atcgaacgta ttaacagcaa
      481 aaagaacaaa accctggtta aaaacatccc atacggttcc ggtctggcgg gtggtgttcg
      541 caacgagcgt tggtatcaac gtaatcgtaa caatatcctg gactacgaga agtggtgctt
      601 cgagtggggc gaagaactgt tccgtatttg caagccgggt gcgactgtac tcgtgtttaa
      661 ctctacccgt accgttgctc acgttcaggt tgcgctggaa aacgcgggtt tctatgctcg
      721 tgacatcctg gtatacaagc gcccgtctgg tatcccgaaa ggtgtaaaca ttaaacagca
      781 gctggagaaa aaaggttacg aaaactctga gaaatgggac ggttggcaca gctgcctgcg
      841 taacgaatgg gaagcgatct gcgttctgca aaaaccgctg gtgaacaact atatcgaaac
      901 tctcctggaa tacggtactg gtctcttcta caccaaagaa tacctgggcg gcttccaatc
      961 taacattctg gacggtatcc agcgtgaacg taccgaagac ttcaacgtgc actgcaccgt
     1021 taaaccgatc gcgctgatgc gtaaactggt tgaaatcttc gttccgcgtc tggaagactc
     1081 tatcctgatc gacccgttcg ctggttctgg tactaccctg ctggctgcga aagagctggg
     1141 tatctcttac atcggtatcg agatcgtttc taactacatc gaaatcatca acaagcgtct
     1201 cgacaacttc tctggtctgc aaggcatcat cccgttcttc caatgatact agaaatattt
     1261 tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     1321 tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc
     1381 tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta
     1441 gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg
     1501 ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg
     1561 cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct
     1621 acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca
     1681 ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga
     1801 tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc
     1861 tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg
     1921 ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc
     1981 tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac
     2041 ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg ctagctattt
     2101 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2161 tgcttcaata atattgaaaa aggaagagta tgagggaagc ggtgatcgcc gaagtatcga
     2221 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     2281 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     2341 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     2401 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     2461 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     2521 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     2581 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     2641 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     2701 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     2761 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     2821 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     2881 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     2941 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctaaca attcgttcaa
     3001 gccgaggggc cgcaagatcc ggccacgatg acccggtcgt cggttcaggg cagggtcgtt
     3061 aaatagccgc ttatgtctat tgctggttta ccggtttatt gactaccgga agcagtgtga
     3121 ccgtgtgctt ctcaaatgcc tgaggtttca gcaaaaaacc cctcaagacc cgtttagagg
     3181 ccccaagggg ttatgctagt tattgctcag cggtggcagc agcctaggtt aattaagctg
     3241 cgctagtaga cgagtccatg tgctggcgtt caaatttcgc agcagcggtt tctttaccag
     3301 actcgacaag ctta
//
