LOCUS       sequence-300477-        3644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses M.XbaI DNA methyltransferase in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3644)
  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 3644)
  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.XbaI-p15A-aadA
FEATURES             Location/Qualifiers
     source          1..3644
                     /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      1615..2160
                     /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     2061..2080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2277..2294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2428..2519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2520..3311
                     /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(3482..3529)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3525..3543
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3581..3625)
                     /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 aaccatgaaa aatgcgacca cccaggctcg tgcggtgcag ggtgactgtc tggttgaact
      361 cgcgcgtctg gaagaatgcg aagctgatct cgtttacctg gacccgccgt tcttcaccaa
      421 ccgtcgtcac tcctccatta gccgtgaccg tattaatgct ttctctttcg acgacgcgtg
      481 gggtgatctg ggtgagtatg cagattttat ggaacaacgt ctccgtgagg cgcaccgtgt
      541 gctgaaacat tctggttcta tcttcgttca ctgcgatact tctgcgaact ttctcctgcg
      601 tactctcctc aacaacatct tcggcgcgtc ccagtttcgc agcgaaatca tctggtccta
      661 caagcgctgg tctaactctg ctcgtaacct gctgccagcg catcagacca tcctgttcta
      721 ctctaaatct gacgcgtaca aattcaatgt tctccacggt tcttattctg aaaccactaa
      781 tgtagatcag atcctccagc tgcgccaacg tgacgctgac ggtgttagca aatatgcgac
      841 tgacaaactc ggtaacacca tctatggcac cgaaaagaat ggcgtgccgc tgaacgacgt
      901 ttgggcaatc ccgtttctca atccgaaagc gaaagaacgt accggctatc cgactcagaa
      961 accaattctg ctgctggaac gcatcatcga aatctccacc gatccaggtg acttcatcgt
     1021 tgacccgttc tgtggttctg gtactaccct ggtggcggca gcgatcctgg gccgtcgtgc
     1081 attcggtatc gacacttctc gcgaagcggt tgctctggca aaccgccgta tcgaggaacc
     1141 ggtgcgtacc gattctgcac tcctgaaaaa gggtcgtctg gcatacgcga ctgctaacct
     1201 caaagcactc gcggttctcg aaggtctgga cttcgtgcca gtacaccgta acagcggtat
     1261 tgatgctttt ctgcgctctc cagaaggtga accgctggtt ccagtacgtg tgcaacgtaa
     1321 cggtgaatct atcagcgacg cggctgcttc cctggcgcgt gctgctatta gcaagaaggc
     1381 gaaaaacgca atcctcgttc gtactaatga agagactggt ctgtttagca ccgacgcact
     1441 cccaccgttt atgcacctga ttgagtctac cgcactgcgc attaaatcta gcctggatgc
     1501 gtccctcatc gcgggtgaga ctaaatccgc agcaaacaac tctttcaaac caaaaccgct
     1561 ccgtggctcc gcgtgatact agaaatattt tatctgatta ataagatgat cttcttgaga
     1621 tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg
     1681 tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc
     1741 gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa
     1801 ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt
     1861 tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt
     1921 gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga
     1981 gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag
     2041 cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2101 caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa
     2161 aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga
     2221 aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg
     2281 agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga cgcaccggtg
     2341 cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag tgccaacata
     2401 gtaagccagt atacactccg ctagctattt gtttattttt ctaaatacat tcaaatatgt
     2461 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2521 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     2581 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     2641 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     2701 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     2761 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     2821 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     2881 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     2941 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     3001 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     3061 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     3121 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     3181 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     3241 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     3301 tcggcaaata atgtctaaca attcgttcaa gccgaggggc cgcaagatcc ggccacgatg
     3361 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
     3421 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggtttca
     3481 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
     3541 cggtggcagc agcctaggtt aattaagctg cgctagtaga cgagtccatg tgctggcgtt
     3601 caaatttcgc agcagcggtt tctttaccag actcgacaag ctta
//
