LOCUS       sequence-115551-        9873 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmid for Paph-gusA integration downstream of B. diazoefficiens 
            (B. japonicum) scoI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9873)
  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 9873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-gusA
FEATURES             Location/Qualifiers
     source          1..9873
                     /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(4592..6403)
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             complement(8090..8461)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            8494..8603
                     /note="incP origin of transfer"
     promoter        join(9854..9859,9860..9873,1..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 cagcttcacc gaggctgtag
     4561 ccgacgatgg tgcgccagga gagttgttga ttcattgttt gcctccctgc tgcggttttt
     4621 caccgaagtt catgccagtc cagcgttttt gcagcagaaa agccgccgac ttcggtttgc
     4681 ggtcgcgagt gaagatccct ttcttgttac cgccaacgcg caatatgcct tgcgaggtcg
     4741 caaaatcggc gaaattccat acctgttcac cgacgacggc gctgacgcga tcaaagacgc
     4801 ggtgatacat atccagccat gcacactgat actcttcact ccacatgtcg gtgtacattg
     4861 agtgcagccc ggctaacgta tccacgccgt attcggtgat gataatcggc tgatgcagtt
     4921 tctcctgcca ggccagaagt tctttttcca gtaccttctc tgccgtttcc aaatcgccgc
     4981 tttggacata ccatccgtaa taacggttca ggcacagcac atcaaagaga tcgctgatgg
     5041 tatcggtgtg agcgtcgcag aacattacat tgacgcaggt gatcggacgc gtcgggtcga
     5101 gtttacgcgt tgcttccgcc agtggcgcga aatattcccg tgcaccttgc ggacgggtat
     5161 ccggttcgtt ggcaatactc cacatcacca cgcttgggtg gtttttgtca cgcgctatca
     5221 gctctttaat cgcctgtaag tgcgcttgct gagtttcccc gttgactgcc tcttcgctgt
     5281 acagttcttt cggcttgttg cccgcttcga aaccaatgcc taaagagagg ttaaagccga
     5341 cagcagcagt ttcatcaatc accacgatgc catgttcatc tgcccagtcg agcatctctt
     5401 cagcgtaagg gtaatgcgag gtacggtagg agttggcccc aatccagtcc attaatgcgt
     5461 ggtcgtgcac catcagcacg ttatcgaatc ctttgccacg caagtccgca tcttcatgac
     5521 gaccaaagcc agtaaagtag aacggtttgt ggttaatcag gaactgttcg cccttcactg
     5581 ccactgaccg gatgccgacg cgaagcgggt agatatcaca ctctgtctgg cttttggctg
     5641 tgacgcacag ttcatagaga taaccttcac ccggttgcca gaggtgcgga ttcaccactt
     5701 gcaaagtccc gctagtgcct tgtccagttg caaccacctg ttgatccgca tcacgcagtt
     5761 caacgctgac atcaccattg gccaccacct gccagtcaac agacgcgtgg ttacagtctt
     5821 gcgcgacatg cgtcaccacg gtgatatcgt ccacccaggt gttcggcgtg gtgtagagca
     5881 ttacgctgcg atggatcccg gcatagttaa agaaatcatg gaagtaagac tgctttttct
     5941 tgccgttttc gtcggtaatc accattcccg gcgggatagt ctgccagttc agttcgttgt
     6001 tcacacaaac ggtgatacgt acacttttcc cggcaataac atacggcgtg acatcggctt
     6061 caaatggcgt atagccgccc tgatgctcca tcacttcctg attattgacc cacactttgc
     6121 cgtaatgagt gaccgcatcg aaacgcagca cgatacgctg gcctgcccaa cctttcggta
     6181 taaagacttc gcgctgatac cagacgttgc ccgcataatt acgaatatct gcatcggcga
     6241 actgatcgtt aaaactgcct ggcacagcaa ttgcccggct ttcttgtaac gcgctttccc
     6301 accaacgctg atcaattcca cagttttcgc gatccagact gaatgcccac aggccgtcga
     6361 gttttttgat ttcacgggtt ggggtttcta caggacgtaa cataagggac tcctcattaa
     6421 gataataata ctggtccgga attctagtat cctgtctctt gatcagatct tgatcccctg
     6481 cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc
     6541 ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag
     6601 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     6661 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     6721 ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag
     6781 cgtgcgagct cgaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
     6841 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     6901 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     6961 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     7021 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     7081 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     7141 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     7201 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     7261 gcccttaaac gcctggttgc tacgcctgaa taagtgataa taagcggatg aatggcagaa
     7321 attcgaaagc aaattcgacc cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta
     7381 tgtctattgc tggtttaccg gtttattgac taccggaagc agtgtgaccg tgtgcttctc
     7441 aaatgcctga ggccagtttg ctcaggctct ccccgtggag gtaataattg acgatatgat
     7501 ccagccgacc aggctttcca cgcccgcgtg ccgctccatg tcgttcgcgc ggttctcgga
     7561 aacgcgctgc cgcgtttcgt gattgtcacg ctcaagcccg tagtcccgtt cgagcgtcgc
     7621 gcagaggtca gcgagggcgc ggtaggcccg atacggctca tggatggtgt ttcgggtcgg
     7681 gtgaatcttg ttgatggcga tatggatgtg caggttgtcg gtgtcgtgat gcacggcact
     7741 gacgcgctga tgctcggcga agccaagccc agcgcagatg cggtcctcaa tcgcgcgcaa
     7801 cgtctccgcg tcgggcttct ctcccgcgcg gaagctaacc agcaggtgat aggtcttgtc
     7861 ggcctcggaa cgggtgttgc cgtgctgggt cgccatcacc tcggccatga cagcgggcag
     7921 ggtgtttgcc tcgcagttcg tgacgcgcac gtgacccagg cgctcggtct tgccttgctc
     7981 gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
     8041 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     8101 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     8161 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     8221 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     8281 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     8341 aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     8401 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     8461 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     8521 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     8581 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     8641 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     8701 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt
     8761 ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc
     8821 gagagacgat gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg
     8881 ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg
     8941 aggcggcgtt agcgtccggc tatgcgctcg tcaccatttg ggagcacatg cgggaaacgg
     9001 ggaaggtcaa gttctcctac gagacgttcc gctcgcacgc caggcggcac atcaaggcca
     9061 agcccgccga tgtgcccgca ccgcaggcca aggctgcgga acccgcgccg gcacccaaga
     9121 cgccggagcc acggcggccg aagcaggggg gcaaggctga aaagccggcc cccgctgcgg
     9181 ccccgaccgg cttcaccttc aacccaacac cggacaaaaa ggatctactg taatggcgaa
     9241 aattcacatg gttttgcagg gcaagggcgg ggtcggcaag tcggccatcg ccgcgatcat
     9301 tgcgcagtac aagatggaca aggggcagac acccttgtgc atcgacaccg acccggtgaa
     9361 cgcgacgttc gagggctaca aggccctgaa cgtccgccgg ctgaacatca tggccggcga
     9421 cgaaattaac tcgcgcaact tcgacaccct ggtcgagctg attgcgccga ccaaggatga
     9481 cgtggtgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     9541 tgctggcgcc tatatcgccg acatcaccga tggggaagat ccttgaagct gtccctgatg
     9601 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     9661 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     9721 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     9781 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     9841 cgacaggcca tgtttgacag cttatcatcg ata
//
