LOCUS       sequence-287551-        7756 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 7756)
  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 7756)
  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: pOPS0386
FEATURES             Location/Qualifiers
     source          1..7756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1553..1639
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1732..1826
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1925..1944)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1936..2751
                     /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            2902..3010
                     /note="incP origin of transfer"
     CDS             complement(3071..4219)
                     /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      4612..5143
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(7665..7751)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccgaattc ggatccggag catcctccct aatgcgcctt gcgcacggtt gtagtcagag
       61 tccgcggtgg gggtaccctc gagggatccg tttaaacgaa ttcaggcttg gaggatacga
      121 atgaagttcc caaaaaactt catgtttgga tattcttggt ctggtttcca gtttgagatg
      181 ggactgccag gaagtgaagt ggaaagcgac tggtgggtgt gggttcacga caaggagaac
      241 atagcatcag gtctagtaag tggagatcta ccagagaacg gcccagcata ttggcacctc
      301 tataagcaag atcatgacat tgcagaaaag ctaggaatgg attgtattag aggtggcatt
      361 gagtgggcaa gaatttttcc aaagccaaca tttgacgtta aagttgatgt ggaaaaggat
      421 gaagaaggca acataatttc cgtagacgtt ccagagagta caataaaaga gctagagaaa
      481 attgccaaca tggaggccct tgaacattat cgcaagattt actcagactg gaaggagagg
      541 ggcaaaacct tcatattaaa cctctaccac tggcctcttc cattatggat tcatgaccca
      601 attgcagtaa ggaaacttgg cccggatagg gctcctgcag gatggttaga tgagaaaaca
      661 gtggtagagt ttgtgaagtt tgccgccttc gttgcttatc accttgatga cctcgttgac
      721 atgtggagca caatgaacga accaaacgta gtctacaatc aaggttacat taatctacgt
      781 tcaggatttc caccaggata tctaagcttt gaagcagcag aaaaggcaaa attcaactta
      841 attcaggctc acatcggagc atatgatgcc ataaaagagt attcagaaaa atccgtggga
      901 gtgatatacg cctttgcttg gcacgatcct ctagcggagg agtataagga tgaagtagag
      961 gaaatcagaa agaaagacta tgagtttgta acaattctac actcaaaagg aaagctagac
     1021 tggatcggcg taaactacta ctccaggctg gtatatggag ccaaagatgg acacctagtt
     1081 cctttacctg gatatggatt tatgagtgag agaggaggat ttgcaaagtc aggcagacct
     1141 gctagtgact ttggatggga aatgtaccca gagggccttg agaaccttct taagtattta
     1201 aacaatgcct acgagctacc aatgataatt acagagaacg gtatggccga tgcagcagat
     1261 agatacaggc cacactatct cgtaagccat ctaaaggcag tttacaatgc tatgaaagaa
     1321 ggtgctgatg ttagagggta tctccactgg tcactaacag acaactacga atgggcccaa
     1381 gggttcagga tgagatttgg attggtttac gtggatttcg agacaaagaa gagatattta
     1441 aggccaagcg ccctggtatt cagagaaata gccactcaaa aagaaattcc agaagaatta
     1501 gctcacctcg cagacctcaa atttgttaca agaaagtagg cttcccaggc atcaaataaa
     1561 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1621 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     1681 agggtggcgg gcaggacgcc cgccataaac tgccagggaa ttccgctctt ggactcctgt
     1741 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1801 gccgggcgtt ttttattggt gagaatccag gcaattgtgt ctcaaaatct ctgatgttac
     1861 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1921 aatacaaggg gtgttatgag ccatattcag cgtgaaacga gctgtagccg tccgcgtctg
     1981 aacagcaaca tggatgcgga tctgtatggc tataaatggg cgcgtgataa cgtgggtcag
     2041 agcggcgcga ccatttatcg tctgtatggc aaaccggatg cgccggaact gtttctgaaa
     2101 catggcaaag gcagcgtggc gaacgatgtg accgatgaaa tggtgcgtct gaactggctg
     2161 accgaattta tgccgctgcc gaccattaaa cattttattc gcaccccgga tgatgcgtgg
     2221 ctgctgacca ccgcgattcc gggcaaaacc gcgtttcagg tgctggaaga atatccggat
     2281 agcggcgaaa acattgtgga tgcgctggcc gtgtttctgc gtcgtctgca tagcattccg
     2341 gtgtgcaact gcccgtttaa cagcgatcgt gtgtttcgtc tggcccaggc gcagagccgt
     2401 atgaacaacg gcctggtgga tgcgagcgat tttgatgatg aacgtaacgg ctggccggtg
     2461 gaacaggtgt ggaaagaaat gcataaactg ctgccgttta gcccggatag cgtggtgacc
     2521 cacggcgatt ttagcctgga taacctgatt ttcgatgaag gcaaactgat tggctgcatt
     2581 gatgtgggcc gtgtgggcat tgcggatcgt tatcaggatc tggccattct gtggaactgc
     2641 ctgggcgaat ttagcccgag cctgcaaaaa cgtctgtttc agaaatatgg cattgataat
     2701 ccggatatga acaaactgca atttcatctg atgctggatg aatttttcta ataattaatt
     2761 gaactacttt tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata
     2821 tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg
     2881 gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg
     2941 ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg
     3001 aggctggccg taggccggcc gcgatgcagg tggctgctga acccccagcc ggaactgacc
     3061 ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca
     3121 ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc
     3181 gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc
     3241 ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct
     3301 cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca
     3361 ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg
     3421 acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc
     3481 gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct
     3541 ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact
     3601 ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg
     3661 tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga
     3721 ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca
     3781 tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct
     3841 tcgtgtgttt cagcaacgcg gcctgcttgg cttcgctgac ctgttttgcc aggtcctcgc
     3901 cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg
     3961 ccaaacctgc cgcctcctgt tcgagccgac gcgaacgctc cacggcggcc gatggcgcgg
     4021 gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga
     4081 ctatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga
     4141 tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc
     4201 ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccggaatta attccccgga
     4261 tcgatccgtc gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4321 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
     4381 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     4441 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     4501 ccggggtcag caccgtttct gcggactggc tttctacgtg gctgccattt ttggggtgag
     4561 gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt tagcgggccg
     4621 ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc
     4681 agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt
     4741 agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca
     4801 gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca
     4861 aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac
     4921 ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc
     4981 cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg
     5041 tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag
     5101 tgagggccaa gttttccgcg aggtatccac aacgccggcg gccctacatg gctctgctgt
     5161 agtgagtggg ttgcgctccg gcagcggtcc tgatcccccg cagaaaaaaa ggatctcaag
     5221 aagatccttt gatcttttct acattactgc atgagcttgt ggaagtgtgc tgagcataaa
     5281 tgattctcct agctgttctt tgggtaagta cgccatcagg acgttgtgag tggcgcgatt
     5341 tttagcggct gaaatcagcc cttgagcctg tcggcaagtc gcgtcatgag gtccatgcgc
     5401 tcatgcagga tcgccacgac caacgcgggt tcgcccgcac gcggcaggca aaaaacgtag
     5461 tggtgttcgc agcgggccat ccgcagcgcg ggaaagagtt cgctcatgtc cttaaacggg
     5521 ccttcgccgg cggcaagcct ggctatgccc tgttccagct tagcgatata gcggcgcacc
     5581 tgcgccgcgc cccactcccg gcgcgtgtag cggatgatgc cgcgtagatc ggcttcggcc
     5641 tcagccgtga ggatgtaggc cgtcaagcgc gatccccgct gagttcttca tcaagaattt
     5701 cgccgacgct cttggtggac accttgccgg caagcccatc gttgatgcgg ttccccagca
     5761 tggttttcag ttcctgccat gcctgatcgg catcagcgtc accggggaac agacgttcga
     5821 gggcgtattg cttaatggtc ttgccctgca aggcggccag ggctttcagg ctctggtgct
     5881 gctggtccgt catgtcgatt gtcaggcggc tcattggata acctccataa aatacacgta
     5941 accacattag cacatatgtg ggcgtgaggc tacagcgcga ggcgcattaa ggtcgggaaa
     6001 atgcgctagg cgcatttaaa ttgcgtattg ctgtaatgcg ccatgccggc tagactaggc
     6061 ccaaatgggt atacccaatt tgaccaaggg ggacgcgatg agggcggcca agcactaccg
     6121 acaacttcta tccatcgact tcaacatcga ggcgctggcc ttcgtgcctg gacccgacgg
     6181 cacacgcggc cggcgcatcc acgtcctggg gcgcgaggtc cgcgaccggc ccggcctggt
     6241 cgagtacctt tcgccggcgt tcggctcgcg ggtggcgctg gacggctact gcaaggccaa
     6301 tttcgatgca gtgctgcacc tggcgtaccc cgatcatcag caatggggcc acgcatgaag
     6361 cgccgaagct acgccatgct gcgcgccgct gccgcgctgg ccgtcctggt cgttgcctcg
     6421 ccggcatggg ccgagctgcg cggcgaggtc gtgcgcatca tcgacggcga caccatcgac
     6481 gtgctggtag acaagcagcc ggtgcgcgtg cgcctggtgg acattgacgc gccggaaaag
     6541 cggcaagcct tcggcgaacg tgcgcgccag gcgctggccg gcatggtgtt ccgccggcac
     6601 gtcctggtcg acgagaagga caccgaccgt tacggccgca cgctgggcac cgtctgggtc
     6661 aacatggagc tggccagccg gccgccgcag ccgcgcaacg tcaacgccgc gatggttcac
     6721 cagggcatgg cgtgggccta tcgcttccac ggccgcgcgg ccgaccctga aatgctgcgg
     6781 ctcgaacagg aggcgcgagg caagcgcgtc ggcctctggt ccgatccgca cgccgtcgag
     6841 ccgtggaaat ggcgacgcga gagcaacaac cggagggacg aaggttgaag gtcgcccgca
     6901 tctacctgcg cgccagtacg gacgagcaga atcttgaacg ccaggagagc cttgtagcgg
     6961 ccacgcgggc cgccgggtac tacgtcgccg gcatctaccg cgagaaggcg tccggcgcac
     7021 gcgccgaccg gcccgagctg ctgcgcatga tcgcggacct gcaacctggt gaagtcgtcg
     7081 ttgcggagaa gatcgaccgc atcagccgct tgccgttggc cgaggccgag cgcctggttg
     7141 cgtcgatccg ggccaaaggg gccaagctgg ccgtgcctgg cgtggtggac ctgtcggagc
     7201 tggccgccga ggcgaacgga gtggcgaaaa tcgttctgga atccgtccag gacatgcttt
     7261 tgaagctcgc cttgcagatg gcccgcgacg actacgagga tcggcgcgag cgtcaacgtc
     7321 agggtgtcca gttggcgaag gccgccggcc gctacaccgg ccgcaaacgt gacgccggca
     7381 tgcacgaccg catcatcacg cttcgctccg gcggatcgag cattgccaag acggccaagc
     7441 tggtcggatg cagcccgagc caggtcaaac gagtgtgggc ggcctggaac gcgcagcagc
     7501 aaaaataaag ccgggcagtg cccggctttt ctcacctttt cgcgtcccgc agggccgctg
     7561 cgagcgccct acctagatcc tcgctttccc cctcggtgta gtccggccag ggcacgaagg
     7621 gcgcggatgc gaacctgttg agcaggtcag atctagggcg gcggatttgt cctactcagg
     7681 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     7741 cgttttattt gatgcc
//
