LOCUS       sequence-256027-        7175 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 7175)
  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 7175)
  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: pOPS0381
FEATURES             Location/Qualifiers
     source          1..7175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(618..704)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             951..1664
                     /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      1681..1767
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1860..1954
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2053..2072)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2064..2879
                     /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            3030..3138
                     /note="incP origin of transfer"
     CDS             complement(3199..4347)
                     /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      4740..5271
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tgatcgcgga cctgcaacct ggtgaagtcg tcgttgcgga gaagatcgac cgcatcagcc
       61 gcttgccgtt ggccgaggcc gagcgcctgg ttgcgtcgat ccgggccaaa ggggccaagc
      121 tggccgtgcc tggcgtggtg gacctgtcgg agctggccgc cgaggcgaac ggagtggcga
      181 aaatcgttct ggaatccgtc caggacatgc ttttgaagct cgccttgcag atggcccgcg
      241 acgactacga ggatcggcgc gagcgtcaac gtcagggtgt ccagttggcg aaggccgccg
      301 gccgctacac cggccgcaaa cgtgacgccg gcatgcacga ccgcatcatc acgcttcgct
      361 ccggcggatc gagcattgcc aagacggcca agctggtcgg atgcagcccg agccaggtca
      421 aacgagtgtg ggcggcctgg aacgcgcagc agcaaaaata aagccgggca gtgcccggct
      481 tttctcacct tttcgcgtcc cgcagggccg ctgcgagcgc cctacctaga tcctcgcttt
      541 ccccctcggt gtagtccggc cagggcacga agggcgcgga tgcgaacctg ttgagcaggt
      601 cagatctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
      661 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct gccgaattcg
      721 gatccggagc tcgacacgac attgtcgtca cctttgtcgg gtccgcgaca agctttttgt
      781 tcctcgatcc cgtatgtttt cgcctaaata tctgaaacag cagcaaatta tttttcgtta
      841 gctcaatcgg cccacttggc acgagtcttg aaatttattg cgaggcggcg gagctgccgg
      901 ccgcatccgt gcggcataac cacggttgtg aacaaatgaa ggaaggccaa atgcgtaaag
      961 gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg
     1021 gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc
     1081 tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc
     1141 tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct
     1201 tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg
     1261 gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg
     1321 agctgaaagg cattgacttt aaagaagatg gcaatatcct gggccataag ctggaataca
     1381 attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga
     1441 attttaaaat tcgccacaac gttgaggatg gcagcgtgca gctggctgat cactaccagc
     1501 aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc
     1561 aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg
     1621 taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgagct tcccaggcat
     1681 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1741 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     1801 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccagggaatt ccgctcttgg
     1861 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     1921 cggttgccgc cgggcgtttt ttattggtga gaatccaggc aattgtgtct caaaatctct
     1981 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     2041 taaacagtaa tacaaggggt gttatgagcc atattcagcg tgaaacgagc tgtagccgtc
     2101 cgcgtctgaa cagcaacatg gatgcggatc tgtatggcta taaatgggcg cgtgataacg
     2161 tgggtcagag cggcgcgacc atttatcgtc tgtatggcaa accggatgcg ccggaactgt
     2221 ttctgaaaca tggcaaaggc agcgtggcga acgatgtgac cgatgaaatg gtgcgtctga
     2281 actggctgac cgaatttatg ccgctgccga ccattaaaca ttttattcgc accccggatg
     2341 atgcgtggct gctgaccacc gcgattccgg gcaaaaccgc gtttcaggtg ctggaagaat
     2401 atccggatag cggcgaaaac attgtggatg cgctggccgt gtttctgcgt cgtctgcata
     2461 gcattccggt gtgcaactgc ccgtttaaca gcgatcgtgt gtttcgtctg gcccaggcgc
     2521 agagccgtat gaacaacggc ctggtggatg cgagcgattt tgatgatgaa cgtaacggct
     2581 ggccggtgga acaggtgtgg aaagaaatgc ataaactgct gccgtttagc ccggatagcg
     2641 tggtgaccca cggcgatttt agcctggata acctgatttt cgatgaaggc aaactgattg
     2701 gctgcattga tgtgggccgt gtgggcattg cggatcgtta tcaggatctg gccattctgt
     2761 ggaactgcct gggcgaattt agcccgagcc tgcaaaaacg tctgtttcag aaatatggca
     2821 ttgataatcc ggatatgaac aaactgcaat ttcatctgat gctggatgaa tttttctaat
     2881 aattaattga actacttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     2941 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     3001 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     3061 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     3121 gctctgcgag gctggccgta ggccggccgc gatgcaggtg gctgctgaac ccccagccgg
     3181 aactgacccc acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg
     3241 ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt
     3301 cttcacgcgg gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta
     3361 cggctcccgg tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg
     3421 gtacttctcc catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc
     3481 gtcgatcagg acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg
     3541 cagcagcgac accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc
     3601 ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc
     3661 caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt
     3721 cgcgtactcc aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc
     3781 ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc
     3841 gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat
     3901 cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat
     3961 ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggct tcgctgacct gttttgccag
     4021 gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat
     4081 cgacttcgcc aaacctgccg cctcctgttc gagccgacgc gaacgctcca cggcggccga
     4141 tggcgcgggc agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc
     4201 ttgctggact atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg
     4261 gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta
     4321 tgccttccgg tcaaacgtcc gattcattca ccctccttgc gggattgccc cggaattaat
     4381 tccccggatc gatccgtcga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     4441 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     4501 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     4561 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4621 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtggc tgccattttt
     4681 ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta
     4741 gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca
     4801 cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa
     4861 gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc
     4921 tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc
     4981 aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg
     5041 cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc
     5101 gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc
     5161 ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca
     5221 tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc cctacatggc
     5281 tctgctgtag tgagtgggtt gcgctccggc agcggtcctg atcccccgca gaaaaaaagg
     5341 atctcaagaa gatcctttga tcttttctac attactgcat gagcttgtgg aagtgtgctg
     5401 agcataaatg attctcctag ctgttctttg ggtaagtacg ccatcaggac gttgtgagtg
     5461 gcgcgatttt tagcggctga aatcagccct tgagcctgtc ggcaagtcgc gtcatgaggt
     5521 ccatgcgctc atgcaggatc gccacgacca acgcgggttc gcccgcacgc ggcaggcaaa
     5581 aaacgtagtg gtgttcgcag cgggccatcc gcagcgcggg aaagagttcg ctcatgtcct
     5641 taaacgggcc ttcgccggcg gcaagcctgg ctatgccctg ttccagctta gcgatatagc
     5701 ggcgcacctg cgccgcgccc cactcccggc gcgtgtagcg gatgatgccg cgtagatcgg
     5761 cttcggcctc agccgtgagg atgtaggccg tcaagcgcga tccccgctga gttcttcatc
     5821 aagaatttcg ccgacgctct tggtggacac cttgccggca agcccatcgt tgatgcggtt
     5881 ccccagcatg gttttcagtt cctgccatgc ctgatcggca tcagcgtcac cggggaacag
     5941 acgttcgagg gcgtattgct taatggtctt gccctgcaag gcggccaggg ctttcaggct
     6001 ctggtgctgc tggtccgtca tgtcgattgt caggcggctc attggataac ctccataaaa
     6061 tacacgtaac cacattagca catatgtggg cgtgaggcta cagcgcgagg cgcattaagg
     6121 tcgggaaaat gcgctaggcg catttaaatt gcgtattgct gtaatgcgcc atgccggcta
     6181 gactaggccc aaatgggtat acccaatttg accaaggggg acgcgatgag ggcggccaag
     6241 cactaccgac aacttctatc catcgacttc aacatcgagg cgctggcctt cgtgcctgga
     6301 cccgacggca cacgcggccg gcgcatccac gtcctggggc gcgaggtccg cgaccggccc
     6361 ggcctggtcg agtacctttc gccggcgttc ggctcgcggg tggcgctgga cggctactgc
     6421 aaggccaatt tcgatgcagt gctgcacctg gcgtaccccg atcatcagca atggggccac
     6481 gcatgaagcg ccgaagctac gccatgctgc gcgccgctgc cgcgctggcc gtcctggtcg
     6541 ttgcctcgcc ggcatgggcc gagctgcgcg gcgaggtcgt gcgcatcatc gacggcgaca
     6601 ccatcgacgt gctggtagac aagcagccgg tgcgcgtgcg cctggtggac attgacgcgc
     6661 cggaaaagcg gcaagccttc ggcgaacgtg cgcgccaggc gctggccggc atggtgttcc
     6721 gccggcacgt cctggtcgac gagaaggaca ccgaccgtta cggccgcacg ctgggcaccg
     6781 tctgggtcaa catggagctg gccagccggc cgccgcagcc gcgcaacgtc aacgccgcga
     6841 tggttcacca gggcatggcg tgggcctatc gcttccacgg ccgcgcggcc gaccctgaaa
     6901 tgctgcggct cgaacaggag gcgcgaggca agcgcgtcgg cctctggtcc gatccgcacg
     6961 ccgtcgagcc gtggaaatgg cgacgcgaga gcaacaaccg gagggacgaa ggttgaaggt
     7021 cgcccgcatc tacctgcgcg ccagtacgga cgagcagaat cttgaacgcc aggagagcct
     7081 tgtagcggcc acgcgggccg ccgggtacta cgtcgccggc atctaccgcg agaaggcgtc
     7141 cggcgcacgc gccgaccggc ccgagctgct gcgca
//
