LOCUS       sequence-409557-        2530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the BHR1 origin insert with a 
            chloramphenicol resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2530)
  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 2530)
  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-clo1
FEATURES             Location/Qualifiers
     source          1..2530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             912..1574
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      1615..1686
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1709..1736
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1757..2416)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2331..2350
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2417..2519)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ctagactttg cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg
       61 ctctccgggc ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga
      121 cgatggccgc gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg
      181 gacaagctga tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg
      241 ctacccagcc ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc
      301 atcaattttt ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct
      361 tgccggttgg acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct
      421 tggccttgac gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa
      481 gcccgcccgc ctgccccgca ggtcccgagc ctcacggcgg cgagtgcggg ggttccaagg
      541 gggcagcgcc accttgggca aggccgaagg ccgcgcagtc gatcaacaag ccccggaggg
      601 gccacttttt gccggagggg gagccgcgcc gaaggcgtgg gggaaccccg caggggtgcc
      661 cttctttggg caccaaagaa ctagatatag ggcgaaatgc gaaagactta aaaatcaaca
      721 acttaaaaaa ggggggtacg caacagctca ttgcggcacc ccccgcaata gctcattgcg
      781 taggttaaag aaaatctgta attgactgcc acttttacgc aacgcataat tgttgtcgcg
      841 ctgccgaaaa gttgcagctg attgcgcatg gtgccgcaac cgtgcggcac ccctaccgca
      901 tggagataag catggccacg cagtccagag aaatcggcat tcaagccaag aacaagcccg
      961 gtcactgggt gcaaacggaa cgcaaagcgc atgaggcgtg ggccgggctt attgcgagga
     1021 aacccacggc ggcaatgctg ctgcatcacc tcgtggcgca gatgggccac cagaacgccg
     1081 tggtggtcag ccagaagaca ctttccaagc tcatcggacg ttctttgcgg acggtccaat
     1141 acgcagtcaa ggacttggtg gccgagcgct ggatctccgt cgtgaagctc aacggccccg
     1201 gcaccgtgtc ggcctacgtg gtcaatgacc gcgtggcgtg gggccagccc cgcgaccagt
     1261 tgcgcctgtc ggtgttcagt gccgccgtgg tggttgatca cgacgaccag gacgaatcgc
     1321 tgttggggca tggcgacctg cgccgcatcc cgaccctgta tccgggcgag cagcaactac
     1381 cgaccggccc cggcgaggag ccgcccagcc agcccggcat tccgggcatg gaaccagacc
     1441 tgccagcctt gaccgaaacg gaggaatggg aacggcgcgg gcagcagcgc ctgccgatgc
     1501 ccgatgagcc gtgttttctg gacgatggcg agccgttgga gccgccgaca cgggtcacgc
     1561 tgccgcgccg gtagcacttg ggttgcgcag caacccgtaa gtgcgctgtt ccagcaaata
     1621 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1681 gctctctact gagacgagac ctcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     1741 tacctaggag tcaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     1801 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     1861 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     1921 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     1981 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     2041 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     2101 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     2161 cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca
     2221 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     2281 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     2341 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     2401 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     2461 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcag
     2521 gtctcgagcg
//
