LOCUS       sequence-256025-        7327 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 7327)
  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 7327)
  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: pOPS0378
FEATURES             Location/Qualifiers
     source          1..7327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(439..525)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             930..1640
                     /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(1075..1093)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      1654..1740
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1833..1927
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2037..2852
                     /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            3003..3111
                     /note="incP origin of transfer"
     CDS             complement(3172..4320)
                     /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      4713..5244
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 aaaatcgttc tggaatccgt ccaggacatg cttttgaagc tcgccttgca gatggcccgc
       61 gacgactacg aggatcggcg cgagcgtcaa cgtcagggtg tccagttggc gaaggccgcc
      121 ggccgctaca ccggccgcaa acgtgacgcc ggcatgcacg accgcatcat cacgcttcgc
      181 tccggcggat cgagcattgc caagacggcc aagctggtcg gatgcagccc gagccaggtc
      241 aaacgagtgt gggcggcctg gaacgcgcag cagcaaaaat aaagccgggc agtgcccggc
      301 ttttctcacc ttttcgcgtc ccgcagggcc gctgcgagcg ccctacctag atcctcgctt
      361 tccccctcgg tgtagtccgg ccagggcacg aagggcgcgg atgcgaacct gttgagcagg
      421 tcagatctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
      481 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tgccgaattc
      541 ggatccggag aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg
      601 atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa
      661 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt
      721 attcaacaaa gcgacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
      781 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
      841 agccatattc aacgggaaac gtcttgctcg agatctatcg atgcatgcca tggtacccgg
      901 gagctcgaat tcaggcttgg aggatacgaa tggtgagcaa gggcgaggag gataacatgg
      961 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1021 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1081 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     1141 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     1201 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1261 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1321 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa aaccatgggc tgggaggcct
     1381 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     1441 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     1501 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     1561 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     1621 tggacgagct gtacaagtaa gcttcccagg catcaaataa aacgaaaggc tcagtcgaaa
     1681 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1741 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     1801 ccgccataaa ctgccaggga attccgctct tggactcctg ttgatagatc cagtaatgac
     1861 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     1921 tgagaatcca ggcaattgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     1981 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     2041 gccatattca gcgtgaaacg agctgtagcc gtccgcgtct gaacagcaac atggatgcgg
     2101 atctgtatgg ctataaatgg gcgcgtgata acgtgggtca gagcggcgcg accatttatc
     2161 gtctgtatgg caaaccggat gcgccggaac tgtttctgaa acatggcaaa ggcagcgtgg
     2221 cgaacgatgt gaccgatgaa atggtgcgtc tgaactggct gaccgaattt atgccgctgc
     2281 cgaccattaa acattttatt cgcaccccgg atgatgcgtg gctgctgacc accgcgattc
     2341 cgggcaaaac cgcgtttcag gtgctggaag aatatccgga tagcggcgaa aacattgtgg
     2401 atgcgctggc cgtgtttctg cgtcgtctgc atagcattcc ggtgtgcaac tgcccgttta
     2461 acagcgatcg tgtgtttcgt ctggcccagg cgcagagccg tatgaacaac ggcctggtgg
     2521 atgcgagcga ttttgatgat gaacgtaacg gctggccggt ggaacaggtg tggaaagaaa
     2581 tgcataaact gctgccgttt agcccggata gcgtggtgac ccacggcgat tttagcctgg
     2641 ataacctgat tttcgatgaa ggcaaactga ttggctgcat tgatgtgggc cgtgtgggca
     2701 ttgcggatcg ttatcaggat ctggccattc tgtggaactg cctgggcgaa tttagcccga
     2761 gcctgcaaaa acgtctgttt cagaaatatg gcattgataa tccggatatg aacaaactgc
     2821 aatttcatct gatgctggat gaatttttct aataattaat tgaactactt ttccgctgca
     2881 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
     2941 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
     3001 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
     3061 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc gtaggccggc
     3121 cgcgatgcag gtggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     3181 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     3241 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     3301 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     3361 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     3421 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     3481 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     3541 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     3601 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     3661 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     3721 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     3781 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     3841 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     3901 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     3961 ggcctgcttg gcttcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     4021 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     4081 ttcgagccga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     4141 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg actatcgagc cgacggactg
     4201 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     4261 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     4321 tcaccctcct tgcgggattg ccccggaatt aattccccgg atcgatccgt cgatcttgat
     4381 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
     4441 ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc
     4501 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
     4561 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     4621 tgcggactgg ctttctacgt ggctgccatt tttggggtga ggccgttcgc ggccgagggg
     4681 cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
     4741 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
     4801 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     4861 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     4921 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     4981 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     5041 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     5101 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     5161 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
     5221 gaggtatcca caacgccggc ggccctacat ggctctgctg tagtgagtgg gttgcgctcc
     5281 ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacattactg catgagcttg tggaagtgtg ctgagcataa atgattctcc tagctgttct
     5401 ttgggtaagt acgccatcag gacgttgtga gtggcgcgat ttttagcggc tgaaatcagc
     5461 ccttgagcct gtcggcaagt cgcgtcatga ggtccatgcg ctcatgcagg atcgccacga
     5521 ccaacgcggg ttcgcccgca cgcggcaggc aaaaaacgta gtggtgttcg cagcgggcca
     5581 tccgcagcgc gggaaagagt tcgctcatgt ccttaaacgg gccttcgccg gcggcaagcc
     5641 tggctatgcc ctgttccagc ttagcgatat agcggcgcac ctgcgccgcg ccccactccc
     5701 ggcgcgtgta gcggatgatg ccgcgtagat cggcttcggc ctcagccgtg aggatgtagg
     5761 ccgtcaagcg cgatccccgc tgagttcttc atcaagaatt tcgccgacgc tcttggtgga
     5821 caccttgccg gcaagcccat cgttgatgcg gttccccagc atggttttca gttcctgcca
     5881 tgcctgatcg gcatcagcgt caccggggaa cagacgttcg agggcgtatt gcttaatggt
     5941 cttgccctgc aaggcggcca gggctttcag gctctggtgc tgctggtccg tcatgtcgat
     6001 tgtcaggcgg ctcattggat aacctccata aaatacacgt aaccacatta gcacatatgt
     6061 gggcgtgagg ctacagcgcg aggcgcatta aggtcgggaa aatgcgctag gcgcatttaa
     6121 attgcgtatt gctgtaatgc gccatgccgg ctagactagg cccaaatggg tatacccaat
     6181 ttgaccaagg gggacgcgat gagggcggcc aagcactacc gacaacttct atccatcgac
     6241 ttcaacatcg aggcgctggc cttcgtgcct ggacccgacg gcacacgcgg ccggcgcatc
     6301 cacgtcctgg ggcgcgaggt ccgcgaccgg cccggcctgg tcgagtacct ttcgccggcg
     6361 ttcggctcgc gggtggcgct ggacggctac tgcaaggcca atttcgatgc agtgctgcac
     6421 ctggcgtacc ccgatcatca gcaatggggc cacgcatgaa gcgccgaagc tacgccatgc
     6481 tgcgcgccgc tgccgcgctg gccgtcctgg tcgttgcctc gccggcatgg gccgagctgc
     6541 gcggcgaggt cgtgcgcatc atcgacggcg acaccatcga cgtgctggta gacaagcagc
     6601 cggtgcgcgt gcgcctggtg gacattgacg cgccggaaaa gcggcaagcc ttcggcgaac
     6661 gtgcgcgcca ggcgctggcc ggcatggtgt tccgccggca cgtcctggtc gacgagaagg
     6721 acaccgaccg ttacggccgc acgctgggca ccgtctgggt caacatggag ctggccagcc
     6781 ggccgccgca gccgcgcaac gtcaacgccg cgatggttca ccagggcatg gcgtgggcct
     6841 atcgcttcca cggccgcgcg gccgaccctg aaatgctgcg gctcgaacag gaggcgcgag
     6901 gcaagcgcgt cggcctctgg tccgatccgc acgccgtcga gccgtggaaa tggcgacgcg
     6961 agagcaacaa ccggagggac gaaggttgaa ggtcgcccgc atctacctgc gcgccagtac
     7021 ggacgagcag aatcttgaac gccaggagag ccttgtagcg gccacgcggg ccgccgggta
     7081 ctacgtcgcc ggcatctacc gcgagaaggc gtccggcgca cgcgccgacc ggcccgagct
     7141 gctgcgcatg atcgcggacc tgcaacctgg tgaagtcgtc gttgcggaga agatcgaccg
     7201 catcagccgc ttgccgttgg ccgaggccga gcgcctggtt gcgtcgatcc gggccaaagg
     7261 ggccaagctg gccgtgcctg gcgtggtgga cctgtcggag ctggccgccg aggcgaacgg
     7321 agtggcg
//
