LOCUS       sequence-287549-        8184 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 8184)
  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 8184)
  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: pOPS0385
FEATURES             Location/Qualifiers
     source          1..8184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             156..1967
                     /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"
     terminator      1981..2067
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2160..2254
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2353..2372)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2364..3179
                     /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            3330..3438
                     /note="incP origin of transfer"
     CDS             complement(3499..4647)
                     /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      5040..5571
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(8093..8179)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccgaattc ggatccggag catcctccct aatgcgcact ctgtggtctc aggagcatcc
       61 tccctaatgc gccttgcgca cggttgtagt cagagtccgc ggtgggggta ccctcgaggg
      121 atccgtttaa acgaattcag gcttggagga tacgaatggt ccgtcctgta gaaaccccaa
      181 cccgtgaaat caaaaaactc gacggcctgt gggcattcag tctggatcgc gaaaactgtg
      241 gaattgatca gcgttggtgg gaaagcgcgt tacaagaaag ccgggcaatt gctgtgccag
      301 gcagttttaa cgatcagttc gccgatgcag atattcgtaa ttatgcgggc aacgtccggt
      361 atcagcgcga agtctttata ccgaaaggtt gggcaggcca gcgtatcgtg ctgcgtttcg
      421 atgcggtcac tcattacggc aaagtgtggg tcaataatca ggaagtgatg gagcatcagg
      481 gcggctatac gccatttgaa gccgatgtca cgccgtatgt tattgccggg aaaagtgtac
      541 gtatcaccgt ttgtgtgaac aacgaactga actggcagac tatcccgccg ggaatggtga
      601 ttaccgacga aaacggcaag aaaaagcagt cttacttcca tgatttcttt aactatgccg
      661 gaatccatcg cagcgtaatg ctctacacca cgccgaacac ctgggtggac gatatcaccg
      721 tggtgacgca tgtcgcgcaa gactgtaacc acgcgtctgt tgactggcag gtggtggcca
      781 atggtgatgt cagcgttgaa ctgcgtgatg cggatcaaca ggtggttgca actggacaag
      841 gcactagcgg gactttgcaa gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc
      901 tctatgaact gtgcgtcaca gccaaaagcc agacagagtg tgatatctac ccgcttcgcg
      961 tcggcatccg gtcagtggca gtgaagggcg aacagttcct gattaaccac aaaccgttct
     1021 actttactgg ctttggtcgt catgaagatg cggacttgcg tggcaaagga ttcgataacg
     1081 tgctgatggt gcacgaccac gcattaatgg actggattgg ggccaactcc taccgtacct
     1141 cgcattaccc ttacgctgaa gagatgctcg actgggcaga tgaacatggc atcgtggtga
     1201 ttgatgaaac tgctgctgtc ggctttaacc tctctttagg cattggtttc gaagcgggca
     1261 acaagccgaa agaactgtac agcgaagagg cagtcaacgg ggaaactcag caagcgcact
     1321 tacaggcgat taaagagctg atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga
     1381 gtattgccaa cgaaccggat acccgtccgc aaggtgcacg ggaatatttc gcgccactgg
     1441 cggaagcaac gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct
     1501 gcgacgctca caccgatacc atcagcgatc tctttgatgt gctgtgcctg aaccgttatt
     1561 acggatggta tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac
     1621 ttctggcctg gcaggagaaa ctgcatcagc cgattatcat caccgaatac ggcgtggata
     1681 cgttagccgg gctgcactca atgtacaccg acatgtggag tgaagagtat cagtgtgcat
     1741 ggctggatat gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat
     1801 ggaatttcgc cgattttgcg acctcgcaag gcatattgcg cgttggcggt aacaagaaag
     1861 ggatcttcac tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga
     1921 ctggcatgaa cttcggtgaa aaaccgcagc agggaggcaa acaatgagct tcccaggcat
     1981 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2041 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     2101 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccagggaatt ccgctcttgg
     2161 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     2221 cggttgccgc cgggcgtttt ttattggtga gaatccaggc aattgtgtct caaaatctct
     2281 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     2341 taaacagtaa tacaaggggt gttatgagcc atattcagcg tgaaacgagc tgtagccgtc
     2401 cgcgtctgaa cagcaacatg gatgcggatc tgtatggcta taaatgggcg cgtgataacg
     2461 tgggtcagag cggcgcgacc atttatcgtc tgtatggcaa accggatgcg ccggaactgt
     2521 ttctgaaaca tggcaaaggc agcgtggcga acgatgtgac cgatgaaatg gtgcgtctga
     2581 actggctgac cgaatttatg ccgctgccga ccattaaaca ttttattcgc accccggatg
     2641 atgcgtggct gctgaccacc gcgattccgg gcaaaaccgc gtttcaggtg ctggaagaat
     2701 atccggatag cggcgaaaac attgtggatg cgctggccgt gtttctgcgt cgtctgcata
     2761 gcattccggt gtgcaactgc ccgtttaaca gcgatcgtgt gtttcgtctg gcccaggcgc
     2821 agagccgtat gaacaacggc ctggtggatg cgagcgattt tgatgatgaa cgtaacggct
     2881 ggccggtgga acaggtgtgg aaagaaatgc ataaactgct gccgtttagc ccggatagcg
     2941 tggtgaccca cggcgatttt agcctggata acctgatttt cgatgaaggc aaactgattg
     3001 gctgcattga tgtgggccgt gtgggcattg cggatcgtta tcaggatctg gccattctgt
     3061 ggaactgcct gggcgaattt agcccgagcc tgcaaaaacg tctgtttcag aaatatggca
     3121 ttgataatcc ggatatgaac aaactgcaat ttcatctgat gctggatgaa tttttctaat
     3181 aattaattga actacttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     3241 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     3301 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     3361 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     3421 gctctgcgag gctggccgta ggccggccgc gatgcaggtg gctgctgaac ccccagccgg
     3481 aactgacccc acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg
     3541 ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt
     3601 cttcacgcgg gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta
     3661 cggctcccgg tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg
     3721 gtacttctcc catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc
     3781 gtcgatcagg acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg
     3841 cagcagcgac accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc
     3901 ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc
     3961 caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt
     4021 cgcgtactcc aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc
     4081 ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc
     4141 gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat
     4201 cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat
     4261 ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggct tcgctgacct gttttgccag
     4321 gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat
     4381 cgacttcgcc aaacctgccg cctcctgttc gagccgacgc gaacgctcca cggcggccga
     4441 tggcgcgggc agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc
     4501 ttgctggact atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg
     4561 gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta
     4621 tgccttccgg tcaaacgtcc gattcattca ccctccttgc gggattgccc cggaattaat
     4681 tccccggatc gatccgtcga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     4741 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     4801 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     4861 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4921 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtggc tgccattttt
     4981 ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta
     5041 gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca
     5101 cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa
     5161 gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc
     5221 tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc
     5281 aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg
     5341 cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc
     5401 gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc
     5461 ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca
     5521 tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc cctacatggc
     5581 tctgctgtag tgagtgggtt gcgctccggc agcggtcctg atcccccgca gaaaaaaagg
     5641 atctcaagaa gatcctttga tcttttctac attactgcat gagcttgtgg aagtgtgctg
     5701 agcataaatg attctcctag ctgttctttg ggtaagtacg ccatcaggac gttgtgagtg
     5761 gcgcgatttt tagcggctga aatcagccct tgagcctgtc ggcaagtcgc gtcatgaggt
     5821 ccatgcgctc atgcaggatc gccacgacca acgcgggttc gcccgcacgc ggcaggcaaa
     5881 aaacgtagtg gtgttcgcag cgggccatcc gcagcgcggg aaagagttcg ctcatgtcct
     5941 taaacgggcc ttcgccggcg gcaagcctgg ctatgccctg ttccagctta gcgatatagc
     6001 ggcgcacctg cgccgcgccc cactcccggc gcgtgtagcg gatgatgccg cgtagatcgg
     6061 cttcggcctc agccgtgagg atgtaggccg tcaagcgcga tccccgctga gttcttcatc
     6121 aagaatttcg ccgacgctct tggtggacac cttgccggca agcccatcgt tgatgcggtt
     6181 ccccagcatg gttttcagtt cctgccatgc ctgatcggca tcagcgtcac cggggaacag
     6241 acgttcgagg gcgtattgct taatggtctt gccctgcaag gcggccaggg ctttcaggct
     6301 ctggtgctgc tggtccgtca tgtcgattgt caggcggctc attggataac ctccataaaa
     6361 tacacgtaac cacattagca catatgtggg cgtgaggcta cagcgcgagg cgcattaagg
     6421 tcgggaaaat gcgctaggcg catttaaatt gcgtattgct gtaatgcgcc atgccggcta
     6481 gactaggccc aaatgggtat acccaatttg accaaggggg acgcgatgag ggcggccaag
     6541 cactaccgac aacttctatc catcgacttc aacatcgagg cgctggcctt cgtgcctgga
     6601 cccgacggca cacgcggccg gcgcatccac gtcctggggc gcgaggtccg cgaccggccc
     6661 ggcctggtcg agtacctttc gccggcgttc ggctcgcggg tggcgctgga cggctactgc
     6721 aaggccaatt tcgatgcagt gctgcacctg gcgtaccccg atcatcagca atggggccac
     6781 gcatgaagcg ccgaagctac gccatgctgc gcgccgctgc cgcgctggcc gtcctggtcg
     6841 ttgcctcgcc ggcatgggcc gagctgcgcg gcgaggtcgt gcgcatcatc gacggcgaca
     6901 ccatcgacgt gctggtagac aagcagccgg tgcgcgtgcg cctggtggac attgacgcgc
     6961 cggaaaagcg gcaagccttc ggcgaacgtg cgcgccaggc gctggccggc atggtgttcc
     7021 gccggcacgt cctggtcgac gagaaggaca ccgaccgtta cggccgcacg ctgggcaccg
     7081 tctgggtcaa catggagctg gccagccggc cgccgcagcc gcgcaacgtc aacgccgcga
     7141 tggttcacca gggcatggcg tgggcctatc gcttccacgg ccgcgcggcc gaccctgaaa
     7201 tgctgcggct cgaacaggag gcgcgaggca agcgcgtcgg cctctggtcc gatccgcacg
     7261 ccgtcgagcc gtggaaatgg cgacgcgaga gcaacaaccg gagggacgaa ggttgaaggt
     7321 cgcccgcatc tacctgcgcg ccagtacgga cgagcagaat cttgaacgcc aggagagcct
     7381 tgtagcggcc acgcgggccg ccgggtacta cgtcgccggc atctaccgcg agaaggcgtc
     7441 cggcgcacgc gccgaccggc ccgagctgct gcgcatgatc gcggacctgc aacctggtga
     7501 agtcgtcgtt gcggagaaga tcgaccgcat cagccgcttg ccgttggccg aggccgagcg
     7561 cctggttgcg tcgatccggg ccaaaggggc caagctggcc gtgcctggcg tggtggacct
     7621 gtcggagctg gccgccgagg cgaacggagt ggcgaaaatc gttctggaat ccgtccagga
     7681 catgcttttg aagctcgcct tgcagatggc ccgcgacgac tacgaggatc ggcgcgagcg
     7741 tcaacgtcag ggtgtccagt tggcgaaggc cgccggccgc tacaccggcc gcaaacgtga
     7801 cgccggcatg cacgaccgca tcatcacgct tcgctccggc ggatcgagca ttgccaagac
     7861 ggccaagctg gtcggatgca gcccgagcca ggtcaaacga gtgtgggcgg cctggaacgc
     7921 gcagcagcaa aaataaagcc gggcagtgcc cggcttttct caccttttcg cgtcccgcag
     7981 ggccgctgcg agcgccctac ctagatcctc gctttccccc tcggtgtagt ccggccaggg
     8041 cacgaagggc gcggatgcga acctgttgag caggtcagat ctagggcggc ggatttgtcc
     8101 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     8161 gagcctttcg ttttatttga tgcc
//
