LOCUS       sequence-405087-        1616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the R6K-gamma origin and oriT insert with
            a chloramphenicol resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1616)
  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 1616)
  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-clo4
FEATURES             Location/Qualifiers
     source          1..1616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     529..548
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(615..717)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            888..997
                     /note="incP origin of transfer"
     rep_origin      1112..1500
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      1523..1550
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1571..1616,1..614))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
       61 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
      121 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
      181 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
      241 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
      301 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
      361 catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg
      421 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
      481 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
      541 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
      601 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
      661 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaggt
      721 ctcgagcggt caattcgagg gggatcaatt ccgttttccg ctgcataacc ctgcttcggg
      781 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
      841 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
      901 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
      961 gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag
     1021 ggcaagcgga tggctgatga aaccaagcca accaggaagg gcagcccacc tatcacggaa
     1081 ttgatccggc cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt caacctgttg
     1141 atagtacgta ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc
     1201 tctcatgttt aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta
     1261 ctaagctctc atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat
     1321 cagcaactta aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg
     1381 cttttaaagc ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga
     1441 gaagccctta gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca
     1501 tggggagacg agacctcaca ctggctcacc ttcgggtggg cctttctgcg tttataccta
     1561 ggagtcaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcatta
//
