LOCUS       sequence-405393-        1644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the Bpi ES3 insert with a chloramphenicol
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1644)
  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 1644)
  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-clo28
FEATURES             Location/Qualifiers
     source          1..1644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      706..733
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(754..1413)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
     primer_bind     1328..1347
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1414..1516)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      join(1575..1644,1..319)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc taacgtacta
       61 agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct catgtttgaa
      121 caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag ttttataaga
      181 aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg gacaacaagc
      241 cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg agtgacacag
      301 gaacacttaa cggctgacat gggggtctcg gagacgcttt gcagactgct gctgacacca
      361 gtgtctcctc cattgcgacc atcaacactg gataagtcgc aaattcctct ttcttggcgc
      421 aggctgcctg aagctttctg gcctgctgca aattcctgtt gtcagatcct cccgataatg
      481 cgaactataa tgttacaatc cgatcattat gtttggatcg tttcatattt ctgtttgacc
      541 tgtcaagtat gcaccatcgc aattgcacca attgatgtgg caagctaaac gggggtgaac
      601 ggcaaccgaa atggggaatc agtgagcagg atcagtcgta aagacgaacg gattgccgaa
      661 cttgcggctc ttgttgagga agcgggaatt acgagacctc acactggctc accttcgggt
      721 gggcctttct gcgtttatac ctaggagtca aaattactcc ccgccctgcc actcatcgca
      781 gtactgttgt aattcattaa gcattctacc gacatggaag ccatcacaaa cggcatgatg
      841 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
      901 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
      961 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     1021 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     1081 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     1141 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     1201 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     1261 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     1321 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     1381 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     1441 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     1501 tcttacgtgc cgatcagtca attcgagggg gatcaattcc gtattgatcc ggccacgatg
     1561 cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac gtactaagct
     1621 ctcatgtttc acgtactaag ctct
//
