LOCUS       sequence-115543-        8710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Paph-bjGFP integration downstream of B. diazoefficiens 
            (B. japonicum) scoI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8710)
  AUTHORS   Ledermann R, Bartsch I, Remus-Emsermann M, Vorholt JA, Fischer HM
  TITLE     Stable fluorescent and enzymatic tagging of Bradyrhizobium 
            diazoefficiens to analyze host-plant infection and colonization.
  JOURNAL   Mol Plant Microbe Interact. 2015 Jun 2.
  PUBMED    26035130
REFERENCE   2  (bases 1 to 8710)
  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"; journalName: "Mol Plant 
            Microbe Interact. 2015 Jun 2."
COMMENT     Sequence Label: pRJPaph-bjGFP
FEATURES             Location/Qualifiers
     source          1..8710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             57..1247
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             1888..2079
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2073..2095)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2181..2321
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2236..2255
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2336..2353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2507..3095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2587..2606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4951..4970
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(6927..7298)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7331..7440
                     /note="incP origin of transfer"
     promoter        join(8691..8696,8697..8710,1..3,4..9)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 agctttaatg cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga
       61 aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag
      121 gcttggttat gccggtactg ccgggcctct tgcgggatat cgtccattcc gacagcatcg
      181 ccagtcacta tggcgtgctg ctagcgctat atgcgttgat gcaatttcta tgcgcacccg
      241 ttctcggagc actgtccgac cgctttggcc gccgcccagt cctgctcgct tcgctacttg
      301 gagccactat cgactacgcg atcatggcga ccacacccgt cctgtggatc ctctacgccg
      361 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
      421 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
      481 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
      541 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
      601 aggagtcgca taagggagag cgtcgaccga tgcccttgag agccttcaac ccagtcagct
      661 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
      721 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
      781 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
      841 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
      901 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
      961 tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     1021 ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg
     1081 ctcttaccag cctaacttcg atcactggac cgctgatcgt cacggcgatt tatgccgcct
     1141 cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc
     1201 tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg
     1261 gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac
     1321 tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag
     1381 ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct
     1441 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     1501 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     1561 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     1621 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     1681 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     1741 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     1801 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     1861 aaattccccc ttacacggag gcatcaagtg accaaacagg aaaaaaccgc ccttaacatg
     1921 gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg
     1981 gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc
     2041 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2101 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2161 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     2221 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2281 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     2341 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2401 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2941 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3301 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3361 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3421 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3481 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3541 gttcgccagt taatagtttg cgcaacgttg ttgccattgc cgcggttcgg tgatgccacg
     3601 atcctcgccc tgctggcgaa gatcgaagag aagcaggacg agcttggcaa ggtcatgatg
     3661 ggcgtggtcc gcccgagggc agagccatga ctttttccac cgcggtggcg gccgctctag
     3721 acggtaatag ggatcggaga agtcgacgcg ggcccggagc tgcgggctca ccgccatcga
     3781 ggcgatgata gcatcgcccc ggttcgaggt gagcgcgtcg accagggtct cgaagcggcg
     3841 catctgcacc gtgcaggtga ccttgatctc ctcgcagagc gcgcgggcga gatcgacgtt
     3901 gaagccggcc ggattgccgt cggcgccggt gtagttgaac ggcggatagt cggtctcggt
     3961 caaaaagcgg atcacggtca ggcgcgacag gtccggccgc tccggccgcc gccgcgggtc
     4021 ccagaagccg ggcacggcct gcggggccgc ctggggcgcg acctgcgggg tgacctgggg
     4081 cgccgccgga ggctgcttgg ccggggcctg cgcctgcgcg ccgggcaagc cggcgaacag
     4141 gcaggcgccg acggccagcc ctgtcagcaa gccacgggca gatttggacg atttcgcctg
     4201 ttgcgatgga gccatcggcc ggaaatgaac gaatttcgtt gggatcggag ggccttatag
     4261 accatccgcc cgggaacgcg agcgccgatt gcggtcaggg tgaaggtgcg ctccctctcc
     4321 cgcttgcggg aggggggcgg ggagagggtg tctccgcgga gggacaatcc ccgagaggag
     4381 agagccctca cccggcgctc tgccatagcc gaggctttgc ctcggcgtct tagaggacgg
     4441 ccgccaaaag cggcctatac ctctcccgca agcgggagag gttacaccga ggacgcggtc
     4501 cgaccgatcc accaaactac agataacgcc tgcaagacgt cggtacccaa ttgtcatcac
     4561 ttgtacagct cgtccatgcc gtgggtgatg ccggcggcgg tgacgaactc cagcaggacc
     4621 atgtggtcgc gcttctcgtt cgggtccttc gacagggccg actgggtcga caggtagtgg
     4681 ttgtccggca gcaggaccgg gccgtcgccg atcggggtgt tctgctggta gtggtcggcc
     4741 agctggaccg agccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc
     4801 ttctgcttgt cggcggtgat gtagacgttg tgcgagttgt agttgtactc cagcttgtgg
     4861 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttgaccagg
     4921 gtgtcgccct cgaacttgac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaacgag
     4981 atggtgcgct cctggacgta gccctccggc atggccgact tgaagaagtc gtggcgcttc
     5041 atgtggtccg ggtagcgcga gaagcactgg acgccgtagg tcagggtggt gaccagggtc
     5101 ggccacggga ccggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     5161 gtggcgtcgc cctcgccctc gcccgagacc gagaacttgt ggccgttgac gtcgccgtcc
     5221 agctcgacca ggatcgggac gacgccggtg aacagctcct cgcccttcga cataattata
     5281 tctccttctt atttctagtc aagatcttga tcccctgcgc catcagatcc ttggcggcaa
     5341 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     5401 caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc
     5461 actgcaagct acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     5521 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt
     5581 cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt gcgagctcga attccgtatg
     5641 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     5701 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     5761 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     5821 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     5881 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     5941 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     6001 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     6061 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac
     6121 gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg
     6181 tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt
     6241 tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc
     6301 aggctctccc cgtggaggta ataattgacg atatgatcca gccgaccagg ctttccacgc
     6361 ccgcgtgccg ctccatgtcg ttcgcgcggt tctcggaaac gcgctgccgc gtttcgtgat
     6421 tgtcacgctc aagcccgtag tcccgttcga gcgtcgcgca gaggtcagcg agggcgcggt
     6481 aggcccgata cggctcatgg atggtgtttc gggtcgggtg aatcttgttg atggcgatat
     6541 ggatgtgcag gttgtcggtg tcgtgatgca cggcactgac gcgctgatgc tcggcgaagc
     6601 caagcccagc gcagatgcgg tcctcaatcg cgcgcaacgt ctccgcgtcg ggcttctctc
     6661 ccgcgcggaa gctaaccagc aggtgatagg tcttgtcggc ctcggaacgg gtgttgccgt
     6721 gctgggtcgc catcacctcg gccatgacag cgggcagggt gtttgcctcg cagttcgtga
     6781 cgcgcacgtg acccaggcgc tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct
     6841 ccgcgaagtc gctcttcttg atggagcgca tggggacgtg cttggcaatc acgcgcaccc
     6901 cccggccgtt ttagcggcta aaaaagtcat ggctctgccc tcgggcggac cacgcccatc
     6961 atgaccttgc caagctcgtc ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg
     7021 gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc
     7081 aggcggccca ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg
     7141 cccgtgattt tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc
     7201 gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt
     7261 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     7321 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     7381 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     7441 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     7501 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     7561 tatgcagcgg aaaagcgctg cttccctgct gttttgtgga atatctaccg actggaaaca
     7621 ggcaaatgca ggaaattact gaactgaggg gacaggcgag agacgatgcc aaagagctac
     7681 accgacgagc tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg gcgtgatgag
     7741 gctgcggttg cgttcctggc ggtgagggcg gatgtcgagg cggcgttagc gtccggctat
     7801 gcgctcgtca ccatttggga gcacatgcgg gaaacgggga aggtcaagtt ctcctacgag
     7861 acgttccgct cgcacgccag gcggcacatc aaggccaagc ccgccgatgt gcccgcaccg
     7921 caggccaagg ctgcggaacc cgcgccggca cccaagacgc cggagccacg gcggccgaag
     7981 caggggggca aggctgaaaa gccggccccc gctgcggccc cgaccggctt caccttcaac
     8041 ccaacaccgg acaaaaagga tctactgtaa tggcgaaaat tcacatggtt ttgcagggca
     8101 agggcggggt cggcaagtcg gccatcgccg cgatcattgc gcagtacaag atggacaagg
     8161 ggcagacacc cttgtgcatc gacaccgacc cggtgaacgc gacgttcgag ggctacaagg
     8221 ccctgaacgt ccgccggctg aacatcatgg ccggcgacga aattaactcg cgcaacttcg
     8281 acaccctggt cgagctgatt gcgccgacca aggatgacgt ggtgatcctc tacgccggac
     8341 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     8401 tcaccgatgg ggaagatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     8461 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     8521 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     8581 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     8641 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccatgt ttgacagctt
     8701 atcatcgata
//
