LOCUS       sequence-256023-        7877 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 7877)
  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 7877)
  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: pOPS0262
FEATURES             Location/Qualifiers
     source          1..7877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1674..1760
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1853..1947
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2046..2065)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2057..2872
                     /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            3023..3131
                     /note="incP origin of transfer"
     CDS             complement(3192..4340)
                     /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      4733..5264
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(7786..7872)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccgaattc ggatccggag ctcgacacga cattgtcgtc acctttgtcg ggtccgcgac
       61 aagctttttg ttcctcgatc ccgtatgttt tcgcctaaat atctgaaaca gcagcaaatt
      121 atttttcgtt agctcaatcg gcccacttgg cacgagtctt gaaatttatt gcgaggcggc
      181 ggagctgccg gccgcatccg tgcggcataa ccacggttgt gaacaaatga aggaaggcca
      241 aatgaagttc ccaaaaaact tcatgtttgg atattcttgg tctggtttcc agtttgagat
      301 gggactgcca ggaagtgaag tggaaagcga ctggtgggtg tgggttcacg acaaggagaa
      361 catagcatca ggtctagtaa gtggagatct accagagaac ggcccagcat attggcacct
      421 ctataagcaa gatcatgaca ttgcagaaaa gctaggaatg gattgtatta gaggtggcat
      481 tgagtgggca agaatttttc caaagccaac atttgacgtt aaagttgatg tggaaaagga
      541 tgaagaaggc aacataattt ccgtagacgt tccagagagt acaataaaag agctagagaa
      601 aattgccaac atggaggccc ttgaacatta tcgcaagatt tactcagact ggaaggagag
      661 gggcaaaacc ttcatattaa acctctacca ctggcctctt ccattatgga ttcatgaccc
      721 aattgcagta aggaaacttg gcccggatag ggctcctgca ggatggttag atgagaaaac
      781 agtggtagag tttgtgaagt ttgccgcctt cgttgcttat caccttgatg acctcgttga
      841 catgtggagc acaatgaacg aaccaaacgt agtctacaat caaggttaca ttaatctacg
      901 ttcaggattt ccaccaggat atctaagctt tgaagcagca gaaaaggcaa aattcaactt
      961 aattcaggct cacatcggag catatgatgc cataaaagag tattcagaaa aatccgtggg
     1021 agtgatatac gcctttgctt ggcacgatcc tctagcggag gagtataagg atgaagtaga
     1081 ggaaatcaga aagaaagact atgagtttgt aacaattcta cactcaaaag gaaagctaga
     1141 ctggatcggc gtaaactact actccaggct ggtatatgga gccaaagatg gacacctagt
     1201 tcctttacct ggatatggat ttatgagtga gagaggagga tttgcaaagt caggcagacc
     1261 tgctagtgac tttggatggg aaatgtaccc agagggcctt gagaaccttc ttaagtattt
     1321 aaacaatgcc tacgagctac caatgataat tacagagaac ggtatggccg atgcagcaga
     1381 tagatacagg ccacactatc tcgtaagcca tctaaaggca gtttacaatg ctatgaaaga
     1441 aggtgctgat gttagagggt atctccactg gtcactaaca gacaactacg aatgggccca
     1501 agggttcagg atgagatttg gattggttta cgtggatttc gagacaaaga agagatattt
     1561 aaggccaagc gccctggtat tcagagaaat agccactcaa aaagaaattc cagaagaatt
     1621 agctcacctc gcagacctca aatttgttac aagaaagtag gcttcccagg catcaaataa
     1681 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1741 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     1801 gagggtggcg ggcaggacgc ccgccataaa ctgccaggga attccgctct tggactcctg
     1861 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1921 cgccgggcgt tttttattgg tgagaatcca ggcaattgtg tctcaaaatc tctgatgtta
     1981 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2041 taatacaagg ggtgttatga gccatattca gcgtgaaacg agctgtagcc gtccgcgtct
     2101 gaacagcaac atggatgcgg atctgtatgg ctataaatgg gcgcgtgata acgtgggtca
     2161 gagcggcgcg accatttatc gtctgtatgg caaaccggat gcgccggaac tgtttctgaa
     2221 acatggcaaa ggcagcgtgg cgaacgatgt gaccgatgaa atggtgcgtc tgaactggct
     2281 gaccgaattt atgccgctgc cgaccattaa acattttatt cgcaccccgg atgatgcgtg
     2341 gctgctgacc accgcgattc cgggcaaaac cgcgtttcag gtgctggaag aatatccgga
     2401 tagcggcgaa aacattgtgg atgcgctggc cgtgtttctg cgtcgtctgc atagcattcc
     2461 ggtgtgcaac tgcccgttta acagcgatcg tgtgtttcgt ctggcccagg cgcagagccg
     2521 tatgaacaac ggcctggtgg atgcgagcga ttttgatgat gaacgtaacg gctggccggt
     2581 ggaacaggtg tggaaagaaa tgcataaact gctgccgttt agcccggata gcgtggtgac
     2641 ccacggcgat tttagcctgg ataacctgat tttcgatgaa ggcaaactga ttggctgcat
     2701 tgatgtgggc cgtgtgggca ttgcggatcg ttatcaggat ctggccattc tgtggaactg
     2761 cctgggcgaa tttagcccga gcctgcaaaa acgtctgttt cagaaatatg gcattgataa
     2821 tccggatatg aacaaactgc aatttcatct gatgctggat gaatttttct aataattaat
     2881 tgaactactt ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
     2941 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt
     3001 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
     3061 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
     3121 gaggctggcc gtaggccggc cgcgatgcag gtggctgctg aacccccagc cggaactgac
     3181 cccacaaggc cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc
     3241 aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg
     3301 cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc
     3361 cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc
     3421 tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc
     3481 aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc
     3541 gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg
     3601 cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc
     3661 tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac
     3721 tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag
     3781 gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg
     3841 attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc
     3901 atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc
     3961 ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc caggtcctcg
     4021 ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc
     4081 gccaaacctg ccgcctcctg ttcgagccga cgcgaacgct ccacggcggc cgatggcgcg
     4141 ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg
     4201 actatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg
     4261 atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc
     4321 cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccggaatt aattccccgg
     4381 atcgatccgt cgatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     4441 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     4501 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     4561 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     4621 tccggggtca gcaccgtttc tgcggactgg ctttctacgt ggctgccatt tttggggtga
     4681 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc
     4741 gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg
     4801 cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt
     4861 tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac
     4921 agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc
     4981 aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca
     5041 cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg
     5101 ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct
     5161 gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca
     5221 gtgagggcca agttttccgc gaggtatcca caacgccggc ggccctacat ggctctgctg
     5281 tagtgagtgg gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa
     5341 gaagatcctt tgatcttttc tacattactg catgagcttg tggaagtgtg ctgagcataa
     5401 atgattctcc tagctgttct ttgggtaagt acgccatcag gacgttgtga gtggcgcgat
     5461 ttttagcggc tgaaatcagc ccttgagcct gtcggcaagt cgcgtcatga ggtccatgcg
     5521 ctcatgcagg atcgccacga ccaacgcggg ttcgcccgca cgcggcaggc aaaaaacgta
     5581 gtggtgttcg cagcgggcca tccgcagcgc gggaaagagt tcgctcatgt ccttaaacgg
     5641 gccttcgccg gcggcaagcc tggctatgcc ctgttccagc ttagcgatat agcggcgcac
     5701 ctgcgccgcg ccccactccc ggcgcgtgta gcggatgatg ccgcgtagat cggcttcggc
     5761 ctcagccgtg aggatgtagg ccgtcaagcg cgatccccgc tgagttcttc atcaagaatt
     5821 tcgccgacgc tcttggtgga caccttgccg gcaagcccat cgttgatgcg gttccccagc
     5881 atggttttca gttcctgcca tgcctgatcg gcatcagcgt caccggggaa cagacgttcg
     5941 agggcgtatt gcttaatggt cttgccctgc aaggcggcca gggctttcag gctctggtgc
     6001 tgctggtccg tcatgtcgat tgtcaggcgg ctcattggat aacctccata aaatacacgt
     6061 aaccacatta gcacatatgt gggcgtgagg ctacagcgcg aggcgcatta aggtcgggaa
     6121 aatgcgctag gcgcatttaa attgcgtatt gctgtaatgc gccatgccgg ctagactagg
     6181 cccaaatggg tatacccaat ttgaccaagg gggacgcgat gagggcggcc aagcactacc
     6241 gacaacttct atccatcgac ttcaacatcg aggcgctggc cttcgtgcct ggacccgacg
     6301 gcacacgcgg ccggcgcatc cacgtcctgg ggcgcgaggt ccgcgaccgg cccggcctgg
     6361 tcgagtacct ttcgccggcg ttcggctcgc gggtggcgct ggacggctac tgcaaggcca
     6421 atttcgatgc agtgctgcac ctggcgtacc ccgatcatca gcaatggggc cacgcatgaa
     6481 gcgccgaagc tacgccatgc tgcgcgccgc tgccgcgctg gccgtcctgg tcgttgcctc
     6541 gccggcatgg gccgagctgc gcggcgaggt cgtgcgcatc atcgacggcg acaccatcga
     6601 cgtgctggta gacaagcagc cggtgcgcgt gcgcctggtg gacattgacg cgccggaaaa
     6661 gcggcaagcc ttcggcgaac gtgcgcgcca ggcgctggcc ggcatggtgt tccgccggca
     6721 cgtcctggtc gacgagaagg acaccgaccg ttacggccgc acgctgggca ccgtctgggt
     6781 caacatggag ctggccagcc ggccgccgca gccgcgcaac gtcaacgccg cgatggttca
     6841 ccagggcatg gcgtgggcct atcgcttcca cggccgcgcg gccgaccctg aaatgctgcg
     6901 gctcgaacag gaggcgcgag gcaagcgcgt cggcctctgg tccgatccgc acgccgtcga
     6961 gccgtggaaa tggcgacgcg agagcaacaa ccggagggac gaaggttgaa ggtcgcccgc
     7021 atctacctgc gcgccagtac ggacgagcag aatcttgaac gccaggagag ccttgtagcg
     7081 gccacgcggg ccgccgggta ctacgtcgcc ggcatctacc gcgagaaggc gtccggcgca
     7141 cgcgccgacc ggcccgagct gctgcgcatg atcgcggacc tgcaacctgg tgaagtcgtc
     7201 gttgcggaga agatcgaccg catcagccgc ttgccgttgg ccgaggccga gcgcctggtt
     7261 gcgtcgatcc gggccaaagg ggccaagctg gccgtgcctg gcgtggtgga cctgtcggag
     7321 ctggccgccg aggcgaacgg agtggcgaaa atcgttctgg aatccgtcca ggacatgctt
     7381 ttgaagctcg ccttgcagat ggcccgcgac gactacgagg atcggcgcga gcgtcaacgt
     7441 cagggtgtcc agttggcgaa ggccgccggc cgctacaccg gccgcaaacg tgacgccggc
     7501 atgcacgacc gcatcatcac gcttcgctcc ggcggatcga gcattgccaa gacggccaag
     7561 ctggtcggat gcagcccgag ccaggtcaaa cgagtgtggg cggcctggaa cgcgcagcag
     7621 caaaaataaa gccgggcagt gcccggcttt tctcaccttt tcgcgtcccg cagggccgct
     7681 gcgagcgccc tacctagatc ctcgctttcc ccctcggtgt agtccggcca gggcacgaag
     7741 ggcgcggatg cgaacctgtt gagcaggtca gatctagggc ggcggatttg tcctactcag
     7801 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt
     7861 tcgttttatt tgatgcc
//
