LOCUS       sequence-276541-        8270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid for R. leguminosarum suitable for experiments in 
            environments where there is no antibiotic selection present..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8270)
  AUTHORS   Mendoza-Suarez MA, Geddes BA, Sanchez-Canizares C, Ramirez-Gonzalez 
            RH, Kirchhelle C, Jorrin B, Poole PS
  TITLE     Optimizing Rhizobium-legume symbioses by simultaneous measurement of
            rhizobial competitiveness and N2 fixation in nodules.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 5;117(18):9822-9831. doi: 
            10.1073/pnas.1921225117. Epub 2020 Apr 21.
  PUBMED    32317381
REFERENCE   2  (bases 1 to 8270)
  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: 
            "10.1073/pnas.1921225117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-05-5- 5"; volume: "117"; issue: "18"; pages: "9822-9831"
COMMENT     Sequence Label: pOPS0263
FEATURES             Location/Qualifiers
     source          1..8270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1825..1911
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2004..2098
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2197..2216)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2208..3023
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            3174..3282
                     /note="incP origin of transfer"
     CDS             complement(3343..4491)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      4884..5415
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(7937..8023)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             join(8270,1..1811)
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVRYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
ORIGIN
        1 tggtccgtcc tgtagaaacc ccaacccgtg aaatcaaaaa actcgacggc ctgtgggcat
       61 tcagtctgga tcgcgaaaac tgtggaattg atcagcgttg gtgggaaagc gcgttacaag
      121 aaagccgggc aattgctgtg ccaggcagtt ttaacgatca gttcgccgat gcagatattc
      181 gtaattatgc gggcaacgtc cggtatcagc gcgaagtctt tataccgaaa ggttgggcag
      241 gccagcgtat cgtgctgcgt ttcgatgcgg tcactcatta cggcaaagtg tgggtcaata
      301 atcaggaagt gatggagcat cagggcggct atacgccatt tgaagccgat gtcacgccgt
      361 atgttattgc cgggaaaagt gtacgtatca ccgtttgtgt gaacaacgaa ctgaactggc
      421 agactatccc gccgggaatg gtgattaccg acgaaaacgg caagaaaaag cagtcttact
      481 tccatgattt ctttaactat gccggaatcc atcgcagcgt aatgctctac accacgccga
      541 acacctgggt ggacgatatc accgtggtga cgcatgtcgc gcaagactgt aaccacgcgt
      601 ctgttgactg gcaggtggtg gccaatggtg atgtcagcgt tgaactgcgt gatgcggatc
      661 aacaggtggt tgcaactgga caaggcacta gcgggacttt gcaagtggtg aatccgcacc
      721 tctggcaacc gggtgaaggt tatctctatg aactgtgcgt cacagccaaa agccagacag
      781 agtgtgatat ctacccgctt cgcgtcggca tccggtcagt ggcagtgaag ggcgaacagt
      841 tcctgattaa ccacaaaccg ttctacttta ctggctttgg tcgtcatgaa gatgcggact
      901 tgcgtggcaa aggattcgat aacgtgctga tggtgcacga ccacgcatta atggactgga
      961 ttggggccaa ctcctaccgt acctcgcatt acccttacgc tgaagagatg ctcgactggg
     1021 cagatgaaca tggcatcgtg gtgattgatg aaactgctgc tgtcggcttt aacctctctt
     1081 taggcattgg tttcgaagcg ggcaacaagc cgaaagaact gtacagcgaa gaggcagtca
     1141 acggggaaac tcagcaagcg cacttacagg cgattaaaga gctgatagcg cgtgacaaaa
     1201 accacccaag cgtggtgatg tggagtattg ccaacgaacc ggatacccgt ccgcaaggtg
     1261 cacgggaata tttcgcgcca ctggcggaag caacgcgtaa actcgacccg acgcgtccga
     1321 tcacctgcgt caatgtaatg ttctgcgacg ctcacaccga taccatcagc gatctctttg
     1381 atgtgctgtg cctgaaccgt tattacggat ggtatgtcca aagcggcgat ttggaaacgg
     1441 cagagaaggt actggaaaaa gaacttctgg cctggcagga gaaactgcat cagccgatta
     1501 tcatcaccga atacggcgtg gatacgttag ccgggctgca ctcaatgtac accgacatgt
     1561 ggagtgaaga gtatcagtgt gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca
     1621 gcgccgtcgt cggtgaacag gtatggaatt tcgccgattt tgcgacctcg caaggcatat
     1681 tgcgcgttgg cggtaacaag aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg
     1741 cttttctgct gcaaaaacgc tggactggca tgaacttcgg tgaaaaaccg cagcagggag
     1801 gcaaacaatg agcttcccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     1861 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     1921 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     1981 actgccaggg aattccgctc ttggactcct gttgatagat ccagtaatga cctcagaact
     2041 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     2101 aggcaattgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2161 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2221 agcgtgaaac gagctgtagc cgtccgcgtc tgaacagcaa catggatgcg gatctgtatg
     2281 gctataaatg ggcgcgtgat aacgtgggtc agagcggcgc gaccatttat cgtctgtatg
     2341 gcaaaccgga tgcgccggaa ctgtttctga aacatggcaa aggcagcgtg gcgaacgatg
     2401 tgaccgatga aatggtgcgt ctgaactggc tgaccgaatt tatgccgctg ccgaccatta
     2461 aacattttat tcgcaccccg gatgatgcgt ggctgctgac caccgcgatt ccgggcaaaa
     2521 ccgcgtttca ggtgctggaa gaatatccgg atagcggcga aaacattgtg gatgcgctgg
     2581 ccgtgtttct gcgtcgtctg catagcattc cggtgtgcaa ctgcccgttt aacagcgatc
     2641 gtgtgtttcg tctggcccag gcgcagagcc gtatgaacaa cggcctggtg gatgcgagcg
     2701 attttgatga tgaacgtaac ggctggccgg tggaacaggt gtggaaagaa atgcataaac
     2761 tgctgccgtt tagcccggat agcgtggtga cccacggcga ttttagcctg gataacctga
     2821 ttttcgatga aggcaaactg attggctgca ttgatgtggg ccgtgtgggc attgcggatc
     2881 gttatcagga tctggccatt ctgtggaact gcctgggcga atttagcccg agcctgcaaa
     2941 aacgtctgtt tcagaaatat ggcattgata atccggatat gaacaaactg caatttcatc
     3001 tgatgctgga tgaatttttc taataattaa ttgaactact tttccgctgc ataaccctgc
     3061 ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga
     3121 ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg
     3181 ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac
     3241 ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg ccgcgatgca
     3301 ggtggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac
     3361 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc
     3421 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg
     3481 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg
     3541 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc
     3601 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc
     3661 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc
     3721 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta
     3781 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat
     3841 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc
     3901 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa
     3961 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc
     4021 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa
     4081 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt
     4141 ggcttcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat
     4201 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcgagccg
     4261 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct
     4321 gtcgcgctcg atcttggccg tagcttgctg gactatcgag ccgacggact ggaaggtttc
     4381 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc
     4441 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc
     4501 ttgcgggatt gccccggaat taattccccg gatcgatccg tcgatcttga tcccctgcgc
     4561 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
     4621 ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct
     4681 agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgttttcc
     4741 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     4801 gctttctacg tggctgccat ttttggggtg aggccgttcg cggccgaggg gcgcagcccc
     4861 tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg gggcaccccc
     4921 cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa ggtttataaa
     4981 tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg ggcggaaacc
     5041 cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata ggtgcgcccc
     5101 tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc tgtcagtagt
     5161 cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc acatcatctg
     5221 tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc agctccacgt
     5281 cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg agtcggcccc
     5341 tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg cgaggtatcc
     5401 acaacgccgg cggccctaca tggctctgct gtagtgagtg ggttgcgctc cggcagcggt
     5461 cctgatcccc cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacattact
     5521 gcatgagctt gtggaagtgt gctgagcata aatgattctc ctagctgttc tttgggtaag
     5581 tacgccatca ggacgttgtg agtggcgcga tttttagcgg ctgaaatcag cccttgagcc
     5641 tgtcggcaag tcgcgtcatg aggtccatgc gctcatgcag gatcgccacg accaacgcgg
     5701 gttcgcccgc acgcggcagg caaaaaacgt agtggtgttc gcagcgggcc atccgcagcg
     5761 cgggaaagag ttcgctcatg tccttaaacg ggccttcgcc ggcggcaagc ctggctatgc
     5821 cctgttccag cttagcgata tagcggcgca cctgcgccgc gccccactcc cggcgcgtgt
     5881 agcggatgat gccgcgtaga tcggcttcgg cctcagccgt gaggatgtag gccgtcaagc
     5941 gcgatccccg ctgagttctt catcaagaat ttcgccgacg ctcttggtgg acaccttgcc
     6001 ggcaagccca tcgttgatgc ggttccccag catggttttc agttcctgcc atgcctgatc
     6061 ggcatcagcg tcaccgggga acagacgttc gagggcgtat tgcttaatgg tcttgccctg
     6121 caaggcggcc agggctttca ggctctggtg ctgctggtcc gtcatgtcga ttgtcaggcg
     6181 gctcattgga taacctccat aaaatacacg taaccacatt agcacatatg tgggcgtgag
     6241 gctacagcgc gaggcgcatt aaggtcggga aaatgcgcta ggcgcattta aattgcgtat
     6301 tgctgtaatg cgccatgccg gctagactag gcccaaatgg gtatacccaa tttgaccaag
     6361 ggggacgcga tgagggcggc caagcactac cgacaacttc tatccatcga cttcaacatc
     6421 gaggcgctgg ccttcgtgcc tggacccgac ggcacacgcg gccggcgcat ccacgtcctg
     6481 gggcgcgagg tccgcgaccg gcccggcctg gtcgagtacc tttcgccggc gttcggctcg
     6541 cgggtggcgc tggacggcta ctgcaaggcc aatttcgatg cagtgctgca cctggcgtac
     6601 cccgatcatc agcaatgggg ccacgcatga agcgccgaag ctacgccatg ctgcgcgccg
     6661 ctgccgcgct ggccgtcctg gtcgttgcct cgccggcatg ggccgagctg cgcggcgagg
     6721 tcgtgcgcat catcgacggc gacaccatcg acgtgctggt agacaagcag ccggtgcgcg
     6781 tgcgcctggt ggacattgac gcgccggaaa agcggcaagc cttcggcgaa cgtgcgcgcc
     6841 aggcgctggc cggcatggtg ttccgccggc acgtcctggt cgacgagaag gacaccgacc
     6901 gttacggccg cacgctgggc accgtctggg tcaacatgga gctggccagc cggccgccgc
     6961 agccgcgcaa cgtcaacgcc gcgatggttc accagggcat ggcgtgggcc tatcgcttcc
     7021 acggccgcgc ggccgaccct gaaatgctgc ggctcgaaca ggaggcgcga ggcaagcgcg
     7081 tcggcctctg gtccgatccg cacgccgtcg agccgtggaa atggcgacgc gagagcaaca
     7141 accggaggga cgaaggttga aggtcgcccg catctacctg cgcgccagta cggacgagca
     7201 gaatcttgaa cgccaggaga gccttgtagc ggccacgcgg gccgccgggt actacgtcgc
     7261 cggcatctac cgcgagaagg cgtccggcgc acgcgccgac cggcccgagc tgctgcgcat
     7321 gatcgcggac ctgcaacctg gtgaagtcgt cgttgcggag aagatcgacc gcatcagccg
     7381 cttgccgttg gccgaggccg agcgcctggt tgcgtcgatc cgggccaaag gggccaagct
     7441 ggccgtgcct ggcgtggtgg acctgtcgga gctggccgcc gaggcgaacg gagtggcgaa
     7501 aatcgttctg gaatccgtcc aggacatgct tttgaagctc gccttgcaga tggcccgcga
     7561 cgactacgag gatcggcgcg agcgtcaacg tcagggtgtc cagttggcga aggccgccgg
     7621 ccgctacacc ggccgcaaac gtgacgccgg catgcacgac cgcatcatca cgcttcgctc
     7681 cggcggatcg agcattgcca agacggccaa gctggtcgga tgcagcccga gccaggtcaa
     7741 acgagtgtgg gcggcctgga acgcgcagca gcaaaaataa agccgggcag tgcccggctt
     7801 ttctcacctt ttcgcgtccc gcagggccgc tgcgagcgcc ctacctagat cctcgctttc
     7861 cccctcggtg tagtccggcc agggcacgaa gggcgcggat gcgaacctgt tgagcaggtc
     7921 agatctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     7981 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg ccgaattcgg
     8041 atccggagct cgacacgaca ttgtcgtcac ctttgtcggg tccgcgacaa gctttttgtt
     8101 cctcgatccc gtatgttttc gcctaaatat ctgaaacagc agcaaattat ttttcgttag
     8161 ctcaatcggc ccacttggca cgagtcttga aatttattgc gaggcggcgg agctgccggc
     8221 cgcatccgtg cggcataacc acggttgtga acaaatgaag gaaggccaaa
//
