LOCUS       sequence-405084-        1833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the Ppu ES2 insert with a chloramphenicol
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1833)
  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 1833)
  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-clo18
FEATURES             Location/Qualifiers
     source          1..1833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      79..106
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(127..786)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
     primer_bind     701..720
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(787..889)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            1049..1158
                     /note="incP origin of transfer"
     rep_origin      1273..1661
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gccacacccg gcggatgcca gggccggaag ccgcagtatc catccaacga ggaattcgtc
       61 attacgagac ctcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacctaggag
      121 tcaaaattac tccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
      181 accgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
      241 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
      301 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
      361 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
      421 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
      481 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
      541 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
      601 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
      661 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
      721 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
      781 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
      841 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcag tcaattcgag
      901 ggggatcaat tccgttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
      961 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     1021 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     1081 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     1141 ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg atggctgatg
     1201 aaaccaagcc aaccaggaag ggcagcccac ctatcacgga attgatccgg ccacgatgcg
     1261 tccggcgtag aggatctgaa gatcagcagt tcaacctgtt gatagtacgt actaagctct
     1321 catgtttcac gtactaagct ctcatgttta acgtactaag ctctcatgtt taacgaacta
     1381 aaccctcatg gctaacgtac taagctctca tggctaacgt actaagctct catgtttcac
     1441 gtactaagct ctcatgtttg aacaataaaa ttaatataaa tcagcaactt aaatagcctc
     1501 taaggtttta agttttataa gaaaaaaaag aatatataag gcttttaaag cttttaaggt
     1561 ttaacggttg tggacaacaa gccagggatg taacgcactg agaagccctt agagcctctc
     1621 aaagcaattt tgagtgacac aggaacactt aacggctgac atgggggtct cggagaaatg
     1681 tgatccatat cacatattgc ctgcggccgt cgcgccgcaa ggcagggctg caaggcccga
     1741 tttacaaggg gtggcgccgt gatggcattt ttttttggaa aacccctaaa gcaacgtgca
     1801 aacccgacga taaccattac gaaggttctc tag
//
