LOCUS       sequence-422546-        1872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the Hro ES4 insert with a chloramphenicol
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1872)
  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 1872)
  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-clo23
FEATURES             Location/Qualifiers
     source          1..1872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     206..225
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(292..394)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            554..663
                     /note="incP origin of transfer"
     rep_origin      778..1166
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      1456..1483
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1504..1872,1..291))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
ORIGIN
        1 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
       61 cattgccata cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
      121 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
      181 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
      241 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
      301 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
      361 atggtgaaag ttggaacctc ttacgtgccg atcagtcaat tcgaggggga tcaattccgt
      421 tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct
      481 ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc
      541 aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct
      601 tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc
      661 cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc aagccaacca
      721 ggaagggcag cccacctatc acggaattga tccggccacg atgcgtccgg cgtagaggat
      781 ctgaagatca gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact
      841 aagctctcat gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa
      901 cgtactaagc tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat
      961 gtttgaacaa taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt
     1021 tataagaaaa aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac
     1081 aacaagccag ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttgagt
     1141 gacacaggaa cacttaacgg ctgacatggg ggtctcggag agacacgaga acggcatgcc
     1201 ttgcgcatgc cgtttttttt cgtcccgaca aaacgtgttc ttttcgacaa caaatccggc
     1261 atgaatgcgg caaaagccac accaggcccg tccgcggaac gcccggcttg acccaagccc
     1321 ctgatttgat tgtataattt cggacttttc caaaccggca agtgatcgac aatcgggtca
     1381 agaagcagtg cgaccgacga agtggcgatt ggctaccaaa ctactctgag gcagaaaatt
     1441 acgagacctc acactggctc accttcgggt gggcctttct gcgtttatac ctaggagtca
     1501 aaattactcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctacc
     1561 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     1621 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     1681 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     1741 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     1801 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     1861 cgtttcagtt tg
//
