LOCUS       sequence-404754-        2001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the spectinomycin resistance gene insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2001)
  AUTHORS   van Schaik J, Li Z, Cheadle J, Crook N
  TITLE     Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and 
            Identification of Potential Bacterial Chassis for Studying 
            Plant-Microbe Interactions.
  JOURNAL   ACS Synth Biol. 2023 Oct 20;12(10):3030-3040. doi: 
            10.1021/acssynbio.3c00371. Epub 2023 Sep 15.
  PUBMED    37712562
REFERENCE   2  (bases 1 to 2001)
  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"; doi: 
            "10.1021/acssynbio.3c00371"; journalName: "ACS Synth Biol"; date: 
            "2023-10-20- 20"; volume: "12"; issue: "10"; pages: "3030-3040"
COMMENT     Sequence Label: pMRMTK-clo8
FEATURES             Location/Qualifiers
     source          1..2001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      209..597
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      620..647
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(671..1462)
                     /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"
     oriT            join(1986..2001,1..94)
                     /note="incP origin of transfer"
ORIGIN
        1 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
       61 ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc
      121 aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat cacggaattg
      181 atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa cctgttgata
      241 gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt actaagctct
      301 catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc taacgtacta
      361 agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa tataaatcag
      421 caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata tataaggctt
      481 ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac gcactgagaa
      541 gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg gctgacatgg
      601 gggtctcgct cctcacactg gctcaccttc gggtgggcct ttctgcgttt atacctagga
      661 gtcaaaatta ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt
      721 cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc
      781 tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag
      841 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
      901 gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt
      961 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
     1021 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt
     1081 cgatcgtggc tggctcgaag atacccgcaa gaatgtcatt gcgctgccat tctccaaatt
     1141 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
     1201 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
     1261 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
     1321 tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca
     1381 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
     1441 cggcgatcac cgcttccctc ataatgttta actttgtttt agggcgactg ccctgctgcg
     1501 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     1561 tggatgcccg aggcatagac tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg
     1621 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     1681 tacgctactt gcattacagc ttacgaaccg aacaggctta tgtccactgg gttcgtgcct
     1741 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg tagcagcgaa gtcgaggcat
     1801 ttctgtcctg gctggagcgc gagaccgtca attcgagggg gatcaattcc gttttccgct
     1861 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     1921 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     1981 cagccgggca ggataggtga a
//
