LOCUS       sequence-405380-        2336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the YFP 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-clo42
FEATURES             Location/Qualifiers
     source          1..2336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     562..581
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(648..750)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            910..1019
                     /note="incP origin of transfer"
     rep_origin      1134..1522
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             join(1538..1540,1541..1543,1544..2257)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1583..1604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1844..1863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2191..2212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2276..2303
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2324..2336,1..647))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
ORIGIN
        1 tgccactcat cgcagtactg ttgtaattca ttaagcattc taccgacatg gaagccatca
       61 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
      121 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
      181 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
      241 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
      301 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
      361 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
      421 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
      481 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
      541 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
      601 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
      661 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
      721 tgaaagttgg aacctcttac gtgccgatca gtcaattcga gggggatcaa ttccgttttc
      781 cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt
      841 cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg
      901 gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac
      961 tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccggc
     1021 taccgccggc gtaacagatg agggcaagcg gatggctgat gaaaccaagc caaccaggaa
     1081 gggcagccca cctatcacgg aattgatccg gccacgatgc gtccggcgta gaggatctga
     1141 agatcagcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc
     1201 tctcatgttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta
     1261 ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     1321 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
     1381 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
     1441 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
     1501 caggaacact taacggctga catgggggtc tcgattaatg gtgagcaagg gcgaggagct
     1561 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1621 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     1681 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     1741 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1801 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1861 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1921 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1981 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2041 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2101 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     2161 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2221 cgggatcact ctcggcatgg acgagctgta caagtaaggt acgagacctc acactggctc
     2281 accttcgggt gggcctttct gcgtttatac ctaggagtca aaattactcc ccgccc
//
