LOCUS       sequence-405079-        2192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the Ppu ES3 insert with a chloramphenicol
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2192)
  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 2192)
  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-clo19
FEATURES             Location/Qualifiers
     source          1..2192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(46..705)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
     primer_bind     620..639
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(706..808)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            968..1077
                     /note="incP origin of transfer"
     rep_origin      1192..1580
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      join(2190..2192,1..25)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tcaccttcgg gtgggccttt ctgcgtttat acctaggagt caaaattact ccccgccctg
       61 ccactcatcg cagtactgtt gtaattcatt aagcattcta ccgacatgga agccatcaca
      121 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
      181 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
      241 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
      301 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
      361 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
      421 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
      481 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
      541 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
      601 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
      661 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
      721 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
      781 aaagttggaa cctcttacgt gccgatcagt caattcgagg gggatcaatt ccgttttccg
      841 ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg
      901 cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc
      961 gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg
     1021 tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta
     1081 ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca accaggaagg
     1141 gcagcccacc tatcacggaa ttgatccggc cacgatgcgt ccggcgtaga ggatctgaag
     1201 atcagcagtt caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc
     1261 tcatgtttaa cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact
     1321 aagctctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga
     1381 acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa gttttataag
     1441 aaaaaaaaga atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag
     1501 ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca
     1561 ggaacactta acggctgaca tgggggtctc ggagatcagg gcgctggagg tgtatcgggt
     1621 gagtggcgag agcatgacgg ctcatcgcca gcgtcaattc gcggaaagta gcggcgcaga
     1681 cgcaggcgcg ggcgggcatt tgccctatac tgtcgcgagt ttggcaattg cgcctgcgga
     1741 tcgtcacatt ttgcatcagc gaattgcgct acgattttcg cagatgctgg aacagggctt
     1801 cgtcgacgag gtccgatcgc tgcgagccag aagtgacttg cacgccgggc tgccgtctat
     1861 acgggcagtg gggtatcggc aggtctggga ctacctcgac ggcaagctga ctgagaatga
     1921 gatgcgagaa cgcggtatca ttgccactcg acagctagcc aagcggcagt tcacctggtt
     1981 gcgtggctgg cctgatgtgc attggctcga cagcctggcc tgcgacaatc tgtcccgcac
     2041 cttgaaatac cttggggccg tctccatatt gagctgagtc cctgctgata gccgtctatt
     2101 cttgcgaagg ggcggcctga cttatcaatt tttctgattt tctattattt atccttacag
     2161 gagtgcggca tattacgaga cctcacactg gc
//
