LOCUS       sequence-256028-        7169 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 7169)
  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 7169)
  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: pOPS0382
FEATURES             Location/Qualifiers
     source          1..7169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             242..952
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(387..405)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      966..1052
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1145..1239
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1338..1357)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1349..2164
                     /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            2315..2423
                     /note="incP origin of transfer"
     CDS             complement(2484..3632)
                     /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      4025..4556
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(7078..7164)
                     /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 aatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
      301 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
      361 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
      421 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
      481 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
      541 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
      601 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
      661 aatgcagaag aaaaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
      721 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
      781 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
      841 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
      901 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aagcttccca
      961 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1021 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     1081 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg gaattccgct
     1141 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     1201 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggcaattg tgtctcaaaa
     1261 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1321 ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa cgagctgtag
     1381 ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat gggcgcgtga
     1441 taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg atgcgccgga
     1501 actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg aaatggtgcg
     1561 tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta ttcgcacccc
     1621 ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc aggtgctgga
     1681 agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc tgcgtcgtct
     1741 gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc gtctggccca
     1801 ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg atgaacgtaa
     1861 cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt ttagcccgga
     1921 tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg aaggcaaact
     1981 gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg atctggccat
     2041 tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt ttcagaaata
     2101 tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg atgaattttt
     2161 ctaataatta attgaactac ttttccgctg cataaccctg cttcggggtc attatagcga
     2221 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     2281 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     2341 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     2401 atcctgctct gcgaggctgg ccgtaggccg gccgcgatgc aggtggctgc tgaaccccca
     2461 gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag
     2521 gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc
     2581 cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc
     2641 gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc
     2701 ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt
     2761 tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag
     2821 cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc
     2881 gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac
     2941 cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg
     3001 cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg
     3061 acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc
     3121 gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt
     3181 ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc
     3241 ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcttcgct gacctgtttt
     3301 gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg
     3361 gtcatcgact tcgccaaacc tgccgcctcc tgttcgagcc gacgcgaacg ctccacggcg
     3421 gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc
     3481 gtagcttgct ggactatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg
     3541 gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc
     3601 ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccggaa
     3661 ttaattcccc ggatcgatcc gtcgatcttg atcccctgcg ccatcagatc cttggcggca
     3721 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     3781 gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     3841 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     3901 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtggctgcca
     3961 tttttggggt gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg
     4021 cgttagcggg ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac
     4081 gcgcacaggg cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt
     4141 taaaagacag gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt
     4201 ctgcctgtgg acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc
     4261 ccctcaagtg tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa
     4321 taccgcaggg cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat
     4381 caggcgtttt cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc
     4441 tgcccctcat ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc
     4501 cctcatctgt cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccctac
     4561 atggctctgc tgtagtgagt gggttgcgct ccggcagcgg tcctgatccc ccgcagaaaa
     4621 aaaggatctc aagaagatcc tttgatcttt tctacattac tgcatgagct tgtggaagtg
     4681 tgctgagcat aaatgattct cctagctgtt ctttgggtaa gtacgccatc aggacgttgt
     4741 gagtggcgcg atttttagcg gctgaaatca gcccttgagc ctgtcggcaa gtcgcgtcat
     4801 gaggtccatg cgctcatgca ggatcgccac gaccaacgcg ggttcgcccg cacgcggcag
     4861 gcaaaaaacg tagtggtgtt cgcagcgggc catccgcagc gcgggaaaga gttcgctcat
     4921 gtccttaaac gggccttcgc cggcggcaag cctggctatg ccctgttcca gcttagcgat
     4981 atagcggcgc acctgcgccg cgccccactc ccggcgcgtg tagcggatga tgccgcgtag
     5041 atcggcttcg gcctcagccg tgaggatgta ggccgtcaag cgcgatcccc gctgagttct
     5101 tcatcaagaa tttcgccgac gctcttggtg gacaccttgc cggcaagccc atcgttgatg
     5161 cggttcccca gcatggtttt cagttcctgc catgcctgat cggcatcagc gtcaccgggg
     5221 aacagacgtt cgagggcgta ttgcttaatg gtcttgccct gcaaggcggc cagggctttc
     5281 aggctctggt gctgctggtc cgtcatgtcg attgtcaggc ggctcattgg ataacctcca
     5341 taaaatacac gtaaccacat tagcacatat gtgggcgtga ggctacagcg cgaggcgcat
     5401 taaggtcggg aaaatgcgct aggcgcattt aaattgcgta ttgctgtaat gcgccatgcc
     5461 ggctagacta ggcccaaatg ggtataccca atttgaccaa gggggacgcg atgagggcgg
     5521 ccaagcacta ccgacaactt ctatccatcg acttcaacat cgaggcgctg gccttcgtgc
     5581 ctggacccga cggcacacgc ggccggcgca tccacgtcct ggggcgcgag gtccgcgacc
     5641 ggcccggcct ggtcgagtac ctttcgccgg cgttcggctc gcgggtggcg ctggacggct
     5701 actgcaaggc caatttcgat gcagtgctgc acctggcgta ccccgatcat cagcaatggg
     5761 gccacgcatg aagcgccgaa gctacgccat gctgcgcgcc gctgccgcgc tggccgtcct
     5821 ggtcgttgcc tcgccggcat gggccgagct gcgcggcgag gtcgtgcgca tcatcgacgg
     5881 cgacaccatc gacgtgctgg tagacaagca gccggtgcgc gtgcgcctgg tggacattga
     5941 cgcgccggaa aagcggcaag ccttcggcga acgtgcgcgc caggcgctgg ccggcatggt
     6001 gttccgccgg cacgtcctgg tcgacgagaa ggacaccgac cgttacggcc gcacgctggg
     6061 caccgtctgg gtcaacatgg agctggccag ccggccgccg cagccgcgca acgtcaacgc
     6121 cgcgatggtt caccagggca tggcgtgggc ctatcgcttc cacggccgcg cggccgaccc
     6181 tgaaatgctg cggctcgaac aggaggcgcg aggcaagcgc gtcggcctct ggtccgatcc
     6241 gcacgccgtc gagccgtgga aatggcgacg cgagagcaac aaccggaggg acgaaggttg
     6301 aaggtcgccc gcatctacct gcgcgccagt acggacgagc agaatcttga acgccaggag
     6361 agccttgtag cggccacgcg ggccgccggg tactacgtcg ccggcatcta ccgcgagaag
     6421 gcgtccggcg cacgcgccga ccggcccgag ctgctgcgca tgatcgcgga cctgcaacct
     6481 ggtgaagtcg tcgttgcgga gaagatcgac cgcatcagcc gcttgccgtt ggccgaggcc
     6541 gagcgcctgg ttgcgtcgat ccgggccaaa ggggccaagc tggccgtgcc tggcgtggtg
     6601 gacctgtcgg agctggccgc cgaggcgaac ggagtggcga aaatcgttct ggaatccgtc
     6661 caggacatgc ttttgaagct cgccttgcag atggcccgcg acgactacga ggatcggcgc
     6721 gagcgtcaac gtcagggtgt ccagttggcg aaggccgccg gccgctacac cggccgcaaa
     6781 cgtgacgccg gcatgcacga ccgcatcatc acgcttcgct ccggcggatc gagcattgcc
     6841 aagacggcca agctggtcgg atgcagcccg agccaggtca aacgagtgtg ggcggcctgg
     6901 aacgcgcagc agcaaaaata aagccgggca gtgcccggct tttctcacct tttcgcgtcc
     6961 cgcagggccg ctgcgagcgc cctacctaga tcctcgcttt ccccctcggt gtagtccggc
     7021 cagggcacga agggcgcgga tgcgaacctg ttgagcaggt cagatctagg gcggcggatt
     7081 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     7141 cgactgagcc tttcgtttta tttgatgcc
//
