LOCUS       sequence-405377-        2327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the mCherry insert with a chloramphenicol
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2327)
  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 2327)
  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-clo40
FEATURES             Location/Qualifiers
     source          1..2327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     171..190
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(257..359)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            519..628
                     /note="incP origin of transfer"
     rep_origin      743..1131
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             1147..1857
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1292..1310)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1560..1579
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1876..1903
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1924..2327,1..256))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
ORIGIN
        1 cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca
       61 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
      121 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
      181 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
      241 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
      301 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcag
      361 tcaattcgag ggggatcaat tccgttttcc gctgcataac cctgcttcgg ggtcattata
      421 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
      481 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
      541 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
      601 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
      661 atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcacgga attgatccgg
      721 ccacgatgcg tccggcgtag aggatctgaa gatcagcagt tcaacctgtt gatagtacgt
      781 actaagctct catgtttcac gtactaagct ctcatgttta acgtactaag ctctcatgtt
      841 taacgaacta aaccctcatg gctaacgtac taagctctca tggctaacgt actaagctct
      901 catgtttcac gtactaagct ctcatgtttg aacaataaaa ttaatataaa tcagcaactt
      961 aaatagcctc taaggtttta agttttataa gaaaaaaaag aatatataag gcttttaaag
     1021 cttttaaggt ttaacggttg tggacaacaa gccagggatg taacgcactg agaagccctt
     1081 agagcctctc aaagcaattt tgagtgacac aggaacactt aacggctgac atgggggtct
     1141 cgattaatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1201 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1261 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1321 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1381 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1441 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccttg
     1501 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1561 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1621 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1681 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1741 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1801 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaggt
     1861 acgagacctc acactggctc accttcgggt gggcctttct gcgtttatac ctaggagtca
     1921 aaattactcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctacc
     1981 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     2041 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     2101 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     2161 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     2221 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     2281 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactat
//
