LOCUS       sequence-256026-        7662 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 7662)
  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 7662)
  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: pOPS0380
FEATURES             Location/Qualifiers
     source          1..7662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             735..1445
                     /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(880..898)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      1459..1545
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1638..1732
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1831..1850)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1842..2657
                     /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            2808..2916
                     /note="incP origin of transfer"
     CDS             complement(2977..4125)
                     /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      4518..5049
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(7571..7657)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccgaattc ggatccggag tcgatgctga ccgcctgatc gccacaatag acgacatccg
       61 atacgattct ccgcttagcg atctcggctt tatcttccac gcgattaacg ttgaacacat
      121 gcttcggcga attgacggaa cttgaaggag atcacccgct atcgcagcgg ggacttcaac
      181 acgcctgcaa ctggtaaccc gtgcatcgcc gcggaacgag gccgatgggt tcgctgccac
      241 tttgaaatgt cgtgcttctg gagcgagact tatatgaaca tttgccatcg ctcgcgccca
      301 gacgccgcgt caatgacgtc ttcctcgtga agtggatcgc tagccgtggc gtagtcgagt
      361 gacatggcgc ttgatcatcg ccggaaacag ccctttgcgg caagtgatca gaattcgtta
      421 ctgctggtct ggtgatggga tacaggctgt tcgaacaatg gtcctagaag ctacacatgg
      481 ataaatgaga catcctgtcg gctggtcaga tattcgacat tgtcatcacc tttgtcggtt
      541 acgtgacaag cgttcctgtt ctcaatcccg catctttgcg cccacataac tggaatcgca
      601 gtaaattatt gttcgtcatc tctatcggcc tgcttggcac gaatcttgtg agctatagcg
      661 aggcagcgga acggcctccg catccacgtt gcgggataac cgcattgcct cgaacacatg
      721 aaggaacgcc aaaaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
      781 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
      841 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
      901 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
      961 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1021 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1081 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1141 ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt
     1201 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     1261 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1321 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1381 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1441 agtaagcttc ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1501 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     1561 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     1621 agggaattcc gctcttggac tcctgttgat agatccagta atgacctcag aactccatct
     1681 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaggcaa
     1741 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     1801 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcagcgtg
     1861 aaacgagctg tagccgtccg cgtctgaaca gcaacatgga tgcggatctg tatggctata
     1921 aatgggcgcg tgataacgtg ggtcagagcg gcgcgaccat ttatcgtctg tatggcaaac
     1981 cggatgcgcc ggaactgttt ctgaaacatg gcaaaggcag cgtggcgaac gatgtgaccg
     2041 atgaaatggt gcgtctgaac tggctgaccg aatttatgcc gctgccgacc attaaacatt
     2101 ttattcgcac cccggatgat gcgtggctgc tgaccaccgc gattccgggc aaaaccgcgt
     2161 ttcaggtgct ggaagaatat ccggatagcg gcgaaaacat tgtggatgcg ctggccgtgt
     2221 ttctgcgtcg tctgcatagc attccggtgt gcaactgccc gtttaacagc gatcgtgtgt
     2281 ttcgtctggc ccaggcgcag agccgtatga acaacggcct ggtggatgcg agcgattttg
     2341 atgatgaacg taacggctgg ccggtggaac aggtgtggaa agaaatgcat aaactgctgc
     2401 cgtttagccc ggatagcgtg gtgacccacg gcgattttag cctggataac ctgattttcg
     2461 atgaaggcaa actgattggc tgcattgatg tgggccgtgt gggcattgcg gatcgttatc
     2521 aggatctggc cattctgtgg aactgcctgg gcgaatttag cccgagcctg caaaaacgtc
     2581 tgtttcagaa atatggcatt gataatccgg atatgaacaa actgcaattt catctgatgc
     2641 tggatgaatt tttctaataa ttaattgaac tacttttccg ctgcataacc ctgcttcggg
     2701 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
     2761 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
     2821 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     2881 gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccgcga tgcaggtggc
     2941 tgctgaaccc ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc
     3001 atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc
     3061 gacctgctcg cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt
     3121 ttccagcttg agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc
     3181 cgtcggcgac agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa
     3241 cagcacgacg atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc
     3301 caggacgcgg aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt
     3361 gaagcccatc gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg
     3421 gccattgatc gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc
     3481 gccgataggg gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc
     3541 ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac
     3601 cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg
     3661 ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga
     3721 aagctgcatt tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcttc
     3781 gctgacctgt tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc
     3841 tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc tcctgttcga gccgacgcga
     3901 acgctccacg gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg
     3961 ctcgatcttg gccgtagctt gctggactat cgagccgacg gactggaagg tttcgcgggg
     4021 cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc
     4081 gatcagttct tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg
     4141 gattgccccg gaattaattc cccggatcga tccgtcgatc ttgatcccct gcgccatcag
     4201 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     4261 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     4321 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     4381 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     4441 tacgtggctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag cccctggggg
     4501 gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac cccccttcgg
     4561 cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta taaatattgg
     4621 tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga aacccttgca
     4681 aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg cccctcatct
     4741 gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc
     4801 cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca tctgtgggaa
     4861 actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg
     4921 ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt
     4981 gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt atccacaacg
     5041 ccggcggccc tacatggctc tgctgtagtg agtgggttgc gctccggcag cggtcctgat
     5101 cccccgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacat tactgcatga
     5161 gcttgtggaa gtgtgctgag cataaatgat tctcctagct gttctttggg taagtacgcc
     5221 atcaggacgt tgtgagtggc gcgattttta gcggctgaaa tcagcccttg agcctgtcgg
     5281 caagtcgcgt catgaggtcc atgcgctcat gcaggatcgc cacgaccaac gcgggttcgc
     5341 ccgcacgcgg caggcaaaaa acgtagtggt gttcgcagcg ggccatccgc agcgcgggaa
     5401 agagttcgct catgtcctta aacgggcctt cgccggcggc aagcctggct atgccctgtt
     5461 ccagcttagc gatatagcgg cgcacctgcg ccgcgcccca ctcccggcgc gtgtagcgga
     5521 tgatgccgcg tagatcggct tcggcctcag ccgtgaggat gtaggccgtc aagcgcgatc
     5581 cccgctgagt tcttcatcaa gaatttcgcc gacgctcttg gtggacacct tgccggcaag
     5641 cccatcgttg atgcggttcc ccagcatggt tttcagttcc tgccatgcct gatcggcatc
     5701 agcgtcaccg gggaacagac gttcgagggc gtattgctta atggtcttgc cctgcaaggc
     5761 ggccagggct ttcaggctct ggtgctgctg gtccgtcatg tcgattgtca ggcggctcat
     5821 tggataacct ccataaaata cacgtaacca cattagcaca tatgtgggcg tgaggctaca
     5881 gcgcgaggcg cattaaggtc gggaaaatgc gctaggcgca tttaaattgc gtattgctgt
     5941 aatgcgccat gccggctaga ctaggcccaa atgggtatac ccaatttgac caagggggac
     6001 gcgatgaggg cggccaagca ctaccgacaa cttctatcca tcgacttcaa catcgaggcg
     6061 ctggccttcg tgcctggacc cgacggcaca cgcggccggc gcatccacgt cctggggcgc
     6121 gaggtccgcg accggcccgg cctggtcgag tacctttcgc cggcgttcgg ctcgcgggtg
     6181 gcgctggacg gctactgcaa ggccaatttc gatgcagtgc tgcacctggc gtaccccgat
     6241 catcagcaat ggggccacgc atgaagcgcc gaagctacgc catgctgcgc gccgctgccg
     6301 cgctggccgt cctggtcgtt gcctcgccgg catgggccga gctgcgcggc gaggtcgtgc
     6361 gcatcatcga cggcgacacc atcgacgtgc tggtagacaa gcagccggtg cgcgtgcgcc
     6421 tggtggacat tgacgcgccg gaaaagcggc aagccttcgg cgaacgtgcg cgccaggcgc
     6481 tggccggcat ggtgttccgc cggcacgtcc tggtcgacga gaaggacacc gaccgttacg
     6541 gccgcacgct gggcaccgtc tgggtcaaca tggagctggc cagccggccg ccgcagccgc
     6601 gcaacgtcaa cgccgcgatg gttcaccagg gcatggcgtg ggcctatcgc ttccacggcc
     6661 gcgcggccga ccctgaaatg ctgcggctcg aacaggaggc gcgaggcaag cgcgtcggcc
     6721 tctggtccga tccgcacgcc gtcgagccgt ggaaatggcg acgcgagagc aacaaccgga
     6781 gggacgaagg ttgaaggtcg cccgcatcta cctgcgcgcc agtacggacg agcagaatct
     6841 tgaacgccag gagagccttg tagcggccac gcgggccgcc gggtactacg tcgccggcat
     6901 ctaccgcgag aaggcgtccg gcgcacgcgc cgaccggccc gagctgctgc gcatgatcgc
     6961 ggacctgcaa cctggtgaag tcgtcgttgc ggagaagatc gaccgcatca gccgcttgcc
     7021 gttggccgag gccgagcgcc tggttgcgtc gatccgggcc aaaggggcca agctggccgt
     7081 gcctggcgtg gtggacctgt cggagctggc cgccgaggcg aacggagtgg cgaaaatcgt
     7141 tctggaatcc gtccaggaca tgcttttgaa gctcgccttg cagatggccc gcgacgacta
     7201 cgaggatcgg cgcgagcgtc aacgtcaggg tgtccagttg gcgaaggccg ccggccgcta
     7261 caccggccgc aaacgtgacg ccggcatgca cgaccgcatc atcacgcttc gctccggcgg
     7321 atcgagcatt gccaagacgg ccaagctggt cggatgcagc ccgagccagg tcaaacgagt
     7381 gtgggcggcc tggaacgcgc agcagcaaaa ataaagccgg gcagtgcccg gcttttctca
     7441 ccttttcgcg tcccgcaggg ccgctgcgag cgccctacct agatcctcgc tttccccctc
     7501 ggtgtagtcc ggccagggca cgaagggcgc ggatgcgaac ctgttgagca ggtcagatct
     7561 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     7621 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cc
//
