LOCUS       sequence-256029-        7054 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 7054)
  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 7054)
  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: pOPS0383
FEATURES             Location/Qualifiers
     source          1..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(673..759)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             885..1598
                     /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
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1615..1701
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1794..1888
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1987..2006)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1998..2813
                     /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            2964..3072
                     /note="incP origin of transfer"
     CDS             complement(3133..4281)
                     /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      4674..5205
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 atctaccgcg agaaggcgtc cggcgcacgc gccgaccggc ccgagctgct gcgcatgatc
       61 gcggacctgc aacctggtga agtcgtcgtt gcggagaaga tcgaccgcat cagccgcttg
      121 ccgttggccg aggccgagcg cctggttgcg tcgatccggg ccaaaggggc caagctggcc
      181 gtgcctggcg tggtggacct gtcggagctg gccgccgagg cgaacggagt ggcgaaaatc
      241 gttctggaat ccgtccagga catgcttttg aagctcgcct tgcagatggc ccgcgacgac
      301 tacgaggatc ggcgcgagcg tcaacgtcag ggtgtccagt tggcgaaggc cgccggccgc
      361 tacaccggcc gcaaacgtga cgccggcatg cacgaccgca tcatcacgct tcgctccggc
      421 ggatcgagca ttgccaagac ggccaagctg gtcggatgca gcccgagcca ggtcaaacga
      481 gtgtgggcgg cctggaacgc gcagcagcaa aaataaagcc gggcagtgcc cggcttttct
      541 caccttttcg cgtcccgcag ggccgctgcg agcgccctac ctagatcctc gctttccccc
      601 tcggtgtagt ccggccaggg cacgaagggc gcggatgcga acctgttgag caggtcagat
      661 ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
      721 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctgccga attcggatcc
      781 ggagcatcct ccctaatgcg ccttgcgcac ggttgtagtc agagtccgcg gtgggggtac
      841 cctcgaggga tccgtttaaa cgaattcagg cttggaggat acgaatgcgt aaaggcgaag
      901 agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata
      961 agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt
     1021 tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt
     1081 atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt
     1141 ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt
     1201 acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga
     1261 aaggcattga ctttaaagaa gatggcaata tcctgggcca taagctggaa tacaatttta
     1321 acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta
     1381 aaattcgcca caacgttgag gatggcagcg tgcagctggc tgatcactac cagcaaaaca
     1441 ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg
     1501 ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg
     1561 cagcgggcat cacgcatggt atggatgaac tgtacaaatg agcttcccag gcatcaaata
     1621 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1681 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     1741 ggagggtggc gggcaggacg cccgccataa actgccaggg aattccgctc ttggactcct
     1801 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     1861 ccgccgggcg ttttttattg gtgagaatcc aggcaattgt gtctcaaaat ctctgatgtt
     1921 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     1981 gtaatacaag gggtgttatg agccatattc agcgtgaaac gagctgtagc cgtccgcgtc
     2041 tgaacagcaa catggatgcg gatctgtatg gctataaatg ggcgcgtgat aacgtgggtc
     2101 agagcggcgc gaccatttat cgtctgtatg gcaaaccgga tgcgccggaa ctgtttctga
     2161 aacatggcaa aggcagcgtg gcgaacgatg tgaccgatga aatggtgcgt ctgaactggc
     2221 tgaccgaatt tatgccgctg ccgaccatta aacattttat tcgcaccccg gatgatgcgt
     2281 ggctgctgac caccgcgatt ccgggcaaaa ccgcgtttca ggtgctggaa gaatatccgg
     2341 atagcggcga aaacattgtg gatgcgctgg ccgtgtttct gcgtcgtctg catagcattc
     2401 cggtgtgcaa ctgcccgttt aacagcgatc gtgtgtttcg tctggcccag gcgcagagcc
     2461 gtatgaacaa cggcctggtg gatgcgagcg attttgatga tgaacgtaac ggctggccgg
     2521 tggaacaggt gtggaaagaa atgcataaac tgctgccgtt tagcccggat agcgtggtga
     2581 cccacggcga ttttagcctg gataacctga ttttcgatga aggcaaactg attggctgca
     2641 ttgatgtggg ccgtgtgggc attgcggatc gttatcagga tctggccatt ctgtggaact
     2701 gcctgggcga atttagcccg agcctgcaaa aacgtctgtt tcagaaatat ggcattgata
     2761 atccggatat gaacaaactg caatttcatc tgatgctgga tgaatttttc taataattaa
     2821 ttgaactact tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
     2881 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
     2941 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
     3001 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     3061 cgaggctggc cgtaggccgg ccgcgatgca ggtggctgct gaacccccag ccggaactga
     3121 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     3181 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     3241 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     3301 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     3361 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     3421 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     3481 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     3541 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     3601 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     3661 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     3721 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     3781 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     3841 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     3901 cttcgtgtgt ttcagcaacg cggcctgctt ggcttcgctg acctgttttg ccaggtcctc
     3961 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     4021 cgccaaacct gccgcctcct gttcgagccg acgcgaacgc tccacggcgg ccgatggcgc
     4081 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     4141 gactatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     4201 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     4261 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccggaat taattccccg
     4321 gatcgatccg tcgatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     4381 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     4441 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     4501 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     4561 atccggggtc agcaccgttt ctgcggactg gctttctacg tggctgccat ttttggggtg
     4621 aggccgttcg cggccgaggg gcgcagcccc tggggggatg ggaggcccgc gttagcgggc
     4681 cgggagggtt cgagaagggg gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc
     4741 gcagccctgg ttaaaaacaa ggtttataaa tattggttta aaagcaggtt aaaagacagg
     4801 ttagcggtgg ccgaaaaacg ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga
     4861 cagcccctca aatgtcaata ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt
     4921 caaggatcgc gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc
     4981 acttatcccc aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc aggcgttttc
     5041 gccgatttgc gaggctggcc agctccacgt cgccggccga aatcgagcct gcccctcatc
     5101 tgtcaacgcc gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc
     5161 agtgagggcc aagttttccg cgaggtatcc acaacgccgg cggccctaca tggctctgct
     5221 gtagtgagtg ggttgcgctc cggcagcggt cctgatcccc cgcagaaaaa aaggatctca
     5281 agaagatcct ttgatctttt ctacattact gcatgagctt gtggaagtgt gctgagcata
     5341 aatgattctc ctagctgttc tttgggtaag tacgccatca ggacgttgtg agtggcgcga
     5401 tttttagcgg ctgaaatcag cccttgagcc tgtcggcaag tcgcgtcatg aggtccatgc
     5461 gctcatgcag gatcgccacg accaacgcgg gttcgcccgc acgcggcagg caaaaaacgt
     5521 agtggtgttc gcagcgggcc atccgcagcg cgggaaagag ttcgctcatg tccttaaacg
     5581 ggccttcgcc ggcggcaagc ctggctatgc cctgttccag cttagcgata tagcggcgca
     5641 cctgcgccgc gccccactcc cggcgcgtgt agcggatgat gccgcgtaga tcggcttcgg
     5701 cctcagccgt gaggatgtag gccgtcaagc gcgatccccg ctgagttctt catcaagaat
     5761 ttcgccgacg ctcttggtgg acaccttgcc ggcaagccca tcgttgatgc ggttccccag
     5821 catggttttc agttcctgcc atgcctgatc ggcatcagcg tcaccgggga acagacgttc
     5881 gagggcgtat tgcttaatgg tcttgccctg caaggcggcc agggctttca ggctctggtg
     5941 ctgctggtcc gtcatgtcga ttgtcaggcg gctcattgga taacctccat aaaatacacg
     6001 taaccacatt agcacatatg tgggcgtgag gctacagcgc gaggcgcatt aaggtcggga
     6061 aaatgcgcta ggcgcattta aattgcgtat tgctgtaatg cgccatgccg gctagactag
     6121 gcccaaatgg gtatacccaa tttgaccaag ggggacgcga tgagggcggc caagcactac
     6181 cgacaacttc tatccatcga cttcaacatc gaggcgctgg ccttcgtgcc tggacccgac
     6241 ggcacacgcg gccggcgcat ccacgtcctg gggcgcgagg tccgcgaccg gcccggcctg
     6301 gtcgagtacc tttcgccggc gttcggctcg cgggtggcgc tggacggcta ctgcaaggcc
     6361 aatttcgatg cagtgctgca cctggcgtac cccgatcatc agcaatgggg ccacgcatga
     6421 agcgccgaag ctacgccatg ctgcgcgccg ctgccgcgct ggccgtcctg gtcgttgcct
     6481 cgccggcatg ggccgagctg cgcggcgagg tcgtgcgcat catcgacggc gacaccatcg
     6541 acgtgctggt agacaagcag ccggtgcgcg tgcgcctggt ggacattgac gcgccggaaa
     6601 agcggcaagc cttcggcgaa cgtgcgcgcc aggcgctggc cggcatggtg ttccgccggc
     6661 acgtcctggt cgacgagaag gacaccgacc gttacggccg cacgctgggc accgtctggg
     6721 tcaacatgga gctggccagc cggccgccgc agccgcgcaa cgtcaacgcc gcgatggttc
     6781 accagggcat ggcgtgggcc tatcgcttcc acggccgcgc ggccgaccct gaaatgctgc
     6841 ggctcgaaca ggaggcgcga ggcaagcgcg tcggcctctg gtccgatccg cacgccgtcg
     6901 agccgtggaa atggcgacgc gagagcaaca accggaggga cgaaggttga aggtcgcccg
     6961 catctacctg cgcgccagta cggacgagca gaatcttgaa cgccaggaga gccttgtagc
     7021 ggccacgcgg gccgccgggt actacgtcgc cggc
//
