LOCUS       sequence-405251-        2333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the sfGFP insert with a chloramphenicol 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2333)
  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 2333)
  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-clo39
FEATURES             Location/Qualifiers
     source          1..2333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     435..454
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(521..623)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            783..892
                     /note="incP origin of transfer"
     rep_origin      1007..1395
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             1411..2124
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKKRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2146..2173
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2194..2333,1..520))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGE"
ORIGIN
        1 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
       61 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
      121 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
      181 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
      241 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
      301 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
      361 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
      421 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
      481 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
      541 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
      601 tggaacctct tacgtgccga tcagtcaatt cgagggggat caattccgtt ttccgctgca
      661 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
      721 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
      781 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
      841 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
      901 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
      961 ccacctatca cggaattgat ccggccacga tgcgtccggc gtagaggatc tgaagatcag
     1021 cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta agctctcatg
     1081 tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac gtactaagct
     1141 ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg tttgaacaat
     1201 aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt ataagaaaaa
     1261 aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca acaagccagg
     1321 gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg acacaggaac
     1381 acttaacggc tgacatgggg gtctcgatta atgcgtaaag gcgaagagct gttcactggt
     1441 gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt
     1501 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact
     1561 ggtaaactgc cggttccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc
     1621 tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa
     1681 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aaagcgtgcg
     1741 gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt
     1801 aaagaggacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt
     1861 tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac
     1921 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat
     1981 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat
     2041 ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg
     2101 catggtatgg atgaactgta caaatgaggt acgagacctc acactggctc accttcgggt
     2161 gggcctttct gcgtttatac ctaggagtca aaattactcc ccgccctgcc actcatcgca
     2221 gtactgttgt aattcattaa gcattctacc gacatggaag ccatcacaaa cggcatgatg
     2281 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgc
//
