LOCUS       sequence-256030-        7048 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 7048)
  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 7048)
  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: pOPS0384
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             105..815
                     /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(250..268)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      829..915
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1008..1102
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1201..1220)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1212..2027
                     /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            2178..2286
                     /note="incP origin of transfer"
     CDS             complement(2347..3495)
                     /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      3888..4419
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6941..7027)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggagcatcct ccctaatgcg ccttgcgcac ggttgtagtc agagtccgcg gtgggggtac
       61 cctcgaggga tccgtttaaa cgaattcagg cttggaggat acgaatggtg agcaagggcg
      121 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      181 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      241 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      301 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      361 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      421 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
      481 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaaaacca
      541 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
      601 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
      661 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
      721 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
      781 actccaccgg cggcatggac gagctgtaca agtaagcttc ccaggcatca aataaaacga
      841 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      901 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
      961 tggcgggcag gacgcccgcc ataaactgcc agggaattcc gctcttggac tcctgttgat
     1021 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     1081 ggcgtttttt attggtgaga atccaggcaa ttgtgtctca aaatctctga tgttacattg
     1141 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1201 caaggggtgt tatgagccat attcagcgtg aaacgagctg tagccgtccg cgtctgaaca
     1261 gcaacatgga tgcggatctg tatggctata aatgggcgcg tgataacgtg ggtcagagcg
     1321 gcgcgaccat ttatcgtctg tatggcaaac cggatgcgcc ggaactgttt ctgaaacatg
     1381 gcaaaggcag cgtggcgaac gatgtgaccg atgaaatggt gcgtctgaac tggctgaccg
     1441 aatttatgcc gctgccgacc attaaacatt ttattcgcac cccggatgat gcgtggctgc
     1501 tgaccaccgc gattccgggc aaaaccgcgt ttcaggtgct ggaagaatat ccggatagcg
     1561 gcgaaaacat tgtggatgcg ctggccgtgt ttctgcgtcg tctgcatagc attccggtgt
     1621 gcaactgccc gtttaacagc gatcgtgtgt ttcgtctggc ccaggcgcag agccgtatga
     1681 acaacggcct ggtggatgcg agcgattttg atgatgaacg taacggctgg ccggtggaac
     1741 aggtgtggaa agaaatgcat aaactgctgc cgtttagccc ggatagcgtg gtgacccacg
     1801 gcgattttag cctggataac ctgattttcg atgaaggcaa actgattggc tgcattgatg
     1861 tgggccgtgt gggcattgcg gatcgttatc aggatctggc cattctgtgg aactgcctgg
     1921 gcgaatttag cccgagcctg caaaaacgtc tgtttcagaa atatggcatt gataatccgg
     1981 atatgaacaa actgcaattt catctgatgc tggatgaatt tttctaataa ttaattgaac
     2041 tacttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     2101 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     2161 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     2221 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     2281 tggccgtagg ccggccgcga tgcaggtggc tgctgaaccc ccagccggaa ctgaccccac
     2341 aaggccctag cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc
     2401 gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg cgccacttct tcacgcgggt
     2461 ggaatccgat ccgcacatga ggcggaaggt ttccagcttg agcgggtacg gctcccggtg
     2521 cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt acttctccca
     2581 tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt cgatcaggac
     2641 ctggcaacgg gacgttttct tgccacggtc caggacgcgg aagcggtgca gcagcgacac
     2701 cgattccagg tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga
     2761 caggcattcc tcggccttcg tgtaataccg gccattgatc gaccagccca ggtcctggca
     2821 aagctcgtag aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg cgtactccaa
     2881 cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat
     2941 cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt
     3001 cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt
     3061 gtccggccac ggcgcaatat cgaacaagga aagctgcatt tccttgatct gctgcttcgt
     3121 gtgtttcagc aacgcggcct gcttggcttc gctgacctgt tttgccaggt cctcgccggc
     3181 ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa
     3241 acctgccgcc tcctgttcga gccgacgcga acgctccacg gcggccgatg gcgcgggcag
     3301 ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt gctggactat
     3361 cgagccgacg gactggaagg tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt
     3421 ttcggcatcc tcggcggaaa accccgcgtc gatcagttct tgcctgtatg ccttccggtc
     3481 aaacgtccga ttcattcacc ctccttgcgg gattgccccg gaattaattc cccggatcga
     3541 tccgtcgatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
     3601 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc cggttcgctt
     3661 gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc
     3721 tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     3781 ggtcagcacc gtttctgcgg actggctttc tacgtggctg ccatttttgg ggtgaggccg
     3841 ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag
     3901 ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc
     3961 ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg
     4021 gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc
     4081 ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga
     4141 tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat
     4201 ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat
     4261 ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa
     4321 cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag
     4381 ggccaagttt tccgcgaggt atccacaacg ccggcggccc tacatggctc tgctgtagtg
     4441 agtgggttgc gctccggcag cggtcctgat cccccgcaga aaaaaaggat ctcaagaaga
     4501 tcctttgatc ttttctacat tactgcatga gcttgtggaa gtgtgctgag cataaatgat
     4561 tctcctagct gttctttggg taagtacgcc atcaggacgt tgtgagtggc gcgattttta
     4621 gcggctgaaa tcagcccttg agcctgtcgg caagtcgcgt catgaggtcc atgcgctcat
     4681 gcaggatcgc cacgaccaac gcgggttcgc ccgcacgcgg caggcaaaaa acgtagtggt
     4741 gttcgcagcg ggccatccgc agcgcgggaa agagttcgct catgtcctta aacgggcctt
     4801 cgccggcggc aagcctggct atgccctgtt ccagcttagc gatatagcgg cgcacctgcg
     4861 ccgcgcccca ctcccggcgc gtgtagcgga tgatgccgcg tagatcggct tcggcctcag
     4921 ccgtgaggat gtaggccgtc aagcgcgatc cccgctgagt tcttcatcaa gaatttcgcc
     4981 gacgctcttg gtggacacct tgccggcaag cccatcgttg atgcggttcc ccagcatggt
     5041 tttcagttcc tgccatgcct gatcggcatc agcgtcaccg gggaacagac gttcgagggc
     5101 gtattgctta atggtcttgc cctgcaaggc ggccagggct ttcaggctct ggtgctgctg
     5161 gtccgtcatg tcgattgtca ggcggctcat tggataacct ccataaaata cacgtaacca
     5221 cattagcaca tatgtgggcg tgaggctaca gcgcgaggcg cattaaggtc gggaaaatgc
     5281 gctaggcgca tttaaattgc gtattgctgt aatgcgccat gccggctaga ctaggcccaa
     5341 atgggtatac ccaatttgac caagggggac gcgatgaggg cggccaagca ctaccgacaa
     5401 cttctatcca tcgacttcaa catcgaggcg ctggccttcg tgcctggacc cgacggcaca
     5461 cgcggccggc gcatccacgt cctggggcgc gaggtccgcg accggcccgg cctggtcgag
     5521 tacctttcgc cggcgttcgg ctcgcgggtg gcgctggacg gctactgcaa ggccaatttc
     5581 gatgcagtgc tgcacctggc gtaccccgat catcagcaat ggggccacgc atgaagcgcc
     5641 gaagctacgc catgctgcgc gccgctgccg cgctggccgt cctggtcgtt gcctcgccgg
     5701 catgggccga gctgcgcggc gaggtcgtgc gcatcatcga cggcgacacc atcgacgtgc
     5761 tggtagacaa gcagccggtg cgcgtgcgcc tggtggacat tgacgcgccg gaaaagcggc
     5821 aagccttcgg cgaacgtgcg cgccaggcgc tggccggcat ggtgttccgc cggcacgtcc
     5881 tggtcgacga gaaggacacc gaccgttacg gccgcacgct gggcaccgtc tgggtcaaca
     5941 tggagctggc cagccggccg ccgcagccgc gcaacgtcaa cgccgcgatg gttcaccagg
     6001 gcatggcgtg ggcctatcgc ttccacggcc gcgcggccga ccctgaaatg ctgcggctcg
     6061 aacaggaggc gcgaggcaag cgcgtcggcc tctggtccga tccgcacgcc gtcgagccgt
     6121 ggaaatggcg acgcgagagc aacaaccgga gggacgaagg ttgaaggtcg cccgcatcta
     6181 cctgcgcgcc agtacggacg agcagaatct tgaacgccag gagagccttg tagcggccac
     6241 gcgggccgcc gggtactacg tcgccggcat ctaccgcgag aaggcgtccg gcgcacgcgc
     6301 cgaccggccc gagctgctgc gcatgatcgc ggacctgcaa cctggtgaag tcgtcgttgc
     6361 ggagaagatc gaccgcatca gccgcttgcc gttggccgag gccgagcgcc tggttgcgtc
     6421 gatccgggcc aaaggggcca agctggccgt gcctggcgtg gtggacctgt cggagctggc
     6481 cgccgaggcg aacggagtgg cgaaaatcgt tctggaatcc gtccaggaca tgcttttgaa
     6541 gctcgccttg cagatggccc gcgacgacta cgaggatcgg cgcgagcgtc aacgtcaggg
     6601 tgtccagttg gcgaaggccg ccggccgcta caccggccgc aaacgtgacg ccggcatgca
     6661 cgaccgcatc atcacgcttc gctccggcgg atcgagcatt gccaagacgg ccaagctggt
     6721 cggatgcagc ccgagccagg tcaaacgagt gtgggcggcc tggaacgcgc agcagcaaaa
     6781 ataaagccgg gcagtgcccg gcttttctca ccttttcgcg tcccgcaggg ccgctgcgag
     6841 cgccctacct agatcctcgc tttccccctc ggtgtagtcc ggccagggca cgaagggcgc
     6901 ggatgcgaac ctgttgagca ggtcagatct agggcggcgg atttgtccta ctcaggagag
     6961 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     7021 ttatttgatg cctgccgaat tcggatcc
//
