LOCUS       sequence-405082-        1688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the rrnB T1 terminator insert with a 
            chloramphenicol resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1688)
  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 1688)
  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-clo12
FEATURES             Location/Qualifiers
     source          1..1688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     554..573
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(640..742)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            902..1011
                     /note="incP origin of transfer"
     rep_origin      1126..1514
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      1530..1601
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1620..1647
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1668..1688,1..639))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
ORIGIN
        1 atcgcagtac tgttgtaatt cattaagcat tctaccgaca tggaagccat cacaaacggc
       61 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
      121 catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
      181 gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa
      241 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
      301 gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac
      361 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
      421 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
      481 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      541 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      601 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      661 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      721 ggaacctctt acgtgccgat cagtcaattc gagggggatc aattccgttt tccgctgcat
      781 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
      841 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
      901 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
      961 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg
     1021 gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc
     1081 cacctatcac ggaattgatc cggccacgat gcgtccggcg tagaggatct gaagatcagc
     1141 agttcaacct gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt
     1201 ttaacgtact aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc
     1261 tcatggctaa cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata
     1321 aaattaatat aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa
     1381 aagaatatat aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg
     1441 atgtaacgca ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca
     1501 cttaacggct gacatggggg tctcgggtac aaataaaacg aaaggctcag tcgaaagact
     1561 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctcccgaga cctcacactg
     1621 gctcaccttc gggtgggcct ttctgcgttt atacctagga gtcaaaatta ctccccgccc
     1681 tgccactc
//
