LOCUS       sequence-409560-        1842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the ColE1 origin insert with a 
            chloramphenicol resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1842)
  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 1842)
  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-clo3
FEATURES             Location/Qualifiers
     source          1..1842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     382..401
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(468..570)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      749..1337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1238..1257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1491..1508
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      1602..1629
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1650..1842,1..467))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
       61 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
      121 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
      181 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
      241 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
      301 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
      361 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
      421 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
      481 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
      541 tgaaagttgg aacctcttac gtgccgatca ggtctcgagc ggctcgccag tcgattggct
      601 gagctcataa gttcctattc cgaagttccg cgaacgcgta aaggatctag gtgaagatcc
      661 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
      721 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      841 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
      901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1021 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1081 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1141 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1201 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1261 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1321 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1381 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1441 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     1501 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     1561 tttcacaccg catatgctgg atcgagacga gacctcacac tggctcacct tcgggtgggc
     1621 ctttctgcgt ttatacctag gagtcaaaat tacgccccgc cctgccactc atcgcagtac
     1681 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     1741 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     1801 acgggggcga agaagttgtc catattggcc acgtttaaat ca
//
