LOCUS       sequence-115538-        8704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Paph-eBFP2 integration downstream of B. diazoefficiens 
            (B. japonicum) scoI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8704)
  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 8704)
  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-eBFP2
FEATURES             Location/Qualifiers
     source          1..8704
                     /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"
     CDS             complement(join(4548..5261,5262..5264,5265..5267))
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     CDS             complement(6921..7292)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7325..7434
                     /note="incP origin of transfer"
     promoter        join(8685..8690,8691..8704,1..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 cggtacctta cttgtacagc
     4561 tcgtccatgc cgagagtgat cccggcggcg gtgcggaact ccagcaggac catgtgatcg
     4621 cgcttctcgt tggggtcttt gctcagcacg gactgggtgc tcaggtagtg gctgtcgggc
     4681 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
     4741 ctgccgtcct ccacgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
     4801 acggccatga tatagatgtt gtggctgttg aagttgtact ccagcttgtg ccccaggatg
     4861 ttgccgtcct ccttgaagtc gacgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     4921 tcgaacttca cctcggcgcg ggtcttgtag gtgccgtcgt ccttgaagaa gatggtgcgc
     4981 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     5041 gggtagcggg cgaagcactg cacgccgtgg ctcagggtgg tcacgagggt gggccagggc
     5101 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg
     5161 ccctcgccct cgcccctcac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     5221 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccataat tatatctcct
     5281 tcttatttct agtcaagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc
     5341 catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc
     5401 cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca
     5461 agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     5521 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc gcttccttta
     5581 gcagcccttg cgccctgagt gcttgcggca gcgtgcgagc tcgaattccg tatggcaatg
     5641 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     5701 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     5761 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     5821 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     5881 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     5941 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     6001 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     6061 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg ctacgcctga
     6121 ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg
     6181 gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga
     6241 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc
     6301 tccccgtgga ggtaataatt gacgatatga tccagccgac caggctttcc acgcccgcgt
     6361 gccgctccat gtcgttcgcg cggttctcgg aaacgcgctg ccgcgtttcg tgattgtcac
     6421 gctcaagccc gtagtcccgt tcgagcgtcg cgcagaggtc agcgagggcg cggtaggccc
     6481 gatacggctc atggatggtg tttcgggtcg ggtgaatctt gttgatggcg atatggatgt
     6541 gcaggttgtc ggtgtcgtga tgcacggcac tgacgcgctg atgctcggcg aagccaagcc
     6601 cagcgcagat gcggtcctca atcgcgcgca acgtctccgc gtcgggcttc tctcccgcgc
     6661 ggaagctaac cagcaggtga taggtcttgt cggcctcgga acgggtgttg ccgtgctggg
     6721 tcgccatcac ctcggccatg acagcgggca gggtgtttgc ctcgcagttc gtgacgcgca
     6781 cgtgacccag gcgctcggtc ttgccttgct cgtcggtgat gtacttcacc agctccgcga
     6841 agtcgctctt cttgatggag cgcatgggga cgtgcttggc aatcacgcgc accccccggc
     6901 cgttttagcg gctaaaaaag tcatggctct gccctcgggc ggaccacgcc catcatgacc
     6961 ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca gggcgaggat cgtggcatca
     7021 ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg
     7081 cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac gacgcccgtg
     7141 attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc ggccgccgcc
     7201 gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat aggtgggctg
     7261 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     7321 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     7381 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     7441 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     7501 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     7561 gcggaaaagc gctgcttccc tgctgttttg tggaatatct accgactgga aacaggcaaa
     7621 tgcaggaaat tactgaactg aggggacagg cgagagacga tgccaaagag ctacaccgac
     7681 gagctggccg agtgggttga atcccgcgcg gccaagaagc gccggcgtga tgaggctgcg
     7741 gttgcgttcc tggcggtgag ggcggatgtc gaggcggcgt tagcgtccgg ctatgcgctc
     7801 gtcaccattt gggagcacat gcgggaaacg gggaaggtca agttctccta cgagacgttc
     7861 cgctcgcacg ccaggcggca catcaaggcc aagcccgccg atgtgcccgc accgcaggcc
     7921 aaggctgcgg aacccgcgcc ggcacccaag acgccggagc cacggcggcc gaagcagggg
     7981 ggcaaggctg aaaagccggc ccccgctgcg gccccgaccg gcttcacctt caacccaaca
     8041 ccggacaaaa aggatctact gtaatggcga aaattcacat ggttttgcag ggcaagggcg
     8101 gggtcggcaa gtcggccatc gccgcgatca ttgcgcagta caagatggac aaggggcaga
     8161 cacccttgtg catcgacacc gacccggtga acgcgacgtt cgagggctac aaggccctga
     8221 acgtccgccg gctgaacatc atggccggcg acgaaattaa ctcgcgcaac ttcgacaccc
     8281 tggtcgagct gattgcgccg accaaggatg acgtggtgat cctctacgcc ggacgcatcg
     8341 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     8401 atggggaaga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     8461 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     8521 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     8581 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     8641 cgagggcgtg caagattccg aataccgcaa gcgacaggcc atgtttgaca gcttatcatc
     8701 gata
//
