LOCUS       sequence-422544-        2073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the Hro ES6 insert with a chloramphenicol
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2073)
  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 2073)
  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-clo25
FEATURES             Location/Qualifiers
     source          1..2073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..74
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(141..243)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      517..905
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      1506..1533
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1554..2073,1..140))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
ORIGIN
        1 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
       61 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
      121 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
      181 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
      241 tcagtcaatt cgagggggat caattccgtt ttccgctgca taaccctgct tcggggtcat
      301 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
      361 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccctaccgcc ggcgtaacag
      421 atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca
      481 cggaattgat ccggccacga tgcgtccggc gtagaggatc tgaagatcag cagttcaacc
      541 tgttgatagt acgtactaag ctctcatgtt tcacgtacta agctctcatg tttaacgtac
      601 taagctctca tgtttaacga actaaaccct catggctaac gtactaagct ctcatggcta
      661 acgtactaag ctctcatgtt tcacgtacta agctctcatg tttgaacaat aaaattaata
      721 taaatcagca acttaaatag cctctaaggt tttaagtttt ataagaaaaa aaagaatata
      781 taaggctttt aaagctttta aggtttaacg gttgtggaca acaagccagg gatgtaacgc
      841 actgagaagc ccttagagcc tctcaaagca attttgagtg acacaggaac acttaacggc
      901 tgacatgggg gtctcggaga ccattgtttg tactgacaag tggattttga ctggcgatac
      961 tagaaatgcg cctactccgg aaatgccggt gtggcttctt tctgaaagga tgtaacgatg
     1021 gcaacaacaa ttaagtgtgt ggctcgtcgc atggccggtg gtaatggcca tgagcacatc
     1081 agtcacttgt ggtggcagca ggtcgacgac tctggtcgag ttttgtcttc tggcgataac
     1141 acgcgtgagc aaatggtggc gtatatcgag aagcatggac accactctgt ctggtgtcca
     1201 gaccgcaatc cggcagtaca gggtgcctgg gtgcgcgtcc ataacaatgg acgtatcaag
     1261 tatgtgcaga cggtggctga tggcaggact accgacaatc tgctttccct gccgttgcgc
     1321 tgagaagcgc agttttcaat ctttggaggt cctgaaatga gcgataagcg tatttatgtg
     1381 gaacggcgtc cggaaggtga ttatgcagta cggcgtccaa actctcagcg cgcaagtgat
     1441 gttctgccaa ctcaacgcga agcgattgac cgcgcgaagc agctgaaatt acgagacctc
     1501 acactggctc accttcgggt gggcctttct gcgtttatac ctaggagtca aaattactcc
     1561 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctacc gacatggaag
     1621 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     1681 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     1741 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     1801 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     1861 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     1921 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     1981 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     2041 gccggataaa acttgtgctt atttttcttt acg
//
