LOCUS       sequence-405378-        2336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the BFP insert with a chloramphenicol 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2336)
  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 2336)
  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-clo41
FEATURES             Location/Qualifiers
     source          1..2336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(341..343,344..346,347..1060)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(386..407)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     994..1015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1079..1106
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1127..1786)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
     primer_bind     1701..1720
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1787..1889)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            2049..2158
                     /note="incP origin of transfer"
     rep_origin      join(2273..2336,1..325)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 agctctcatg tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac
       61 gtactaagct ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg
      121 tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt
      181 ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca
      241 acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg
      301 acacaggaac acttaacggc tgacatgggg gtctcgatta atggtgagca agggcgagga
      361 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      421 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
      481 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
      541 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      601 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
      661 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      721 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
      781 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
      841 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
      901 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
      961 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1021 cgccgggatc actctcggca tggacgagct gtacaagtaa ggtacgagac ctcacactgg
     1081 ctcaccttcg ggtgggcctt tctgcgttta tacctaggag tcaaaattac tccccgccct
     1141 gccactcatc gcagtactgt tgtaattcat taagcattct accgacatgg aagccatcac
     1201 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     1261 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     1321 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     1381 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     1441 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     1501 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     1561 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     1621 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     1681 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     1741 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     1801 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     1861 gaaagttgga acctcttacg tgccgatcag tcaattcgag ggggatcaat tccgttttcc
     1921 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     1981 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     2041 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     2101 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct
     2161 accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag
     2221 ggcagcccac ctatcacgga attgatccgg ccacgatgcg tccggcgtag aggatctgaa
     2281 gatcagcagt tcaacctgtt gatagtacgt actaagctct catgtttcac gtacta
//
