LOCUS       sequence-115548-        8700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Paph-HcRed integration downstream of B. diazoefficiens 
            (B. japonicum) scoI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8700)
  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 8700)
  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-HcRed
FEATURES             Location/Qualifiers
     source          1..8700
                     /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(4563..5249)
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     CDS             complement(6917..7288)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7321..7430
                     /note="incP origin of transfer"
     promoter        join(8681..8686,8687..8700,1..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 tgcaggacta gtcaagatct aagcttcaat
     4561 tctcagttgg ccttctcggg caggtcgctg taccgggcca cgctggcctc gtacagctcg
     4621 aagtactcgt ccttcttctt ccgcagcatc tggagccgga tgtcggtgaa gtggaagccg
     4681 ggcatggtca gggcgcgcac ggccttcttg ctccggtagc tggtgtagtg gtggcagatc
     4741 aggtgccggt cgcccacctt cagggccatc acgttccggc cgcacagcac gccgttctcg
     4801 gggtacacca cctcggtgct gggctcccag ccgccgctct tgttcttcat cacggggccg
     4861 tcggcgggga agttggtgcc gtgcaccttc accttgtaga tcaggcagtt gccctccagg
     4921 ctggtgtcct ggtgggcggt caggatgccg ccgtcctcgt aggtggtggt tctctcccag
     4981 gtgaagccct cggggaagct ctgcttgaag aagtcgggga tctcggcggt gtggtgcacg
     5041 aaggtcctgc tgccgtactc gcagcagggg gccaggatgt cgaaggcgaa gggcaggggg
     5101 gcgccctcgg tcacgtggat tctcatgctc tgggtgccgg cgaaggggtt gccgtcgccc
     5161 tcgccctcgc acttgaagta gtggccgttc acggtgccct ccatgtacat cttgatgcgc
     5221 atactctcct tcagcaggcc gctcaccata attatatctc cttcttattt ctagtatcct
     5281 gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     5341 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     5401 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     5461 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     5521 atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag
     5581 cccttgcgcc ctgagtgctt gcggcagcgt gcgagctcga attccgtatg gcaatgaaag
     5641 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     5701 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     5761 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     5821 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     5881 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     5941 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     6001 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     6061 cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa
     6121 gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc
     6181 agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac
     6241 cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc
     6301 cgtggaggta ataattgacg atatgatcca gccgaccagg ctttccacgc ccgcgtgccg
     6361 ctccatgtcg ttcgcgcggt tctcggaaac gcgctgccgc gtttcgtgat tgtcacgctc
     6421 aagcccgtag tcccgttcga gcgtcgcgca gaggtcagcg agggcgcggt aggcccgata
     6481 cggctcatgg atggtgtttc gggtcgggtg aatcttgttg atggcgatat ggatgtgcag
     6541 gttgtcggtg tcgtgatgca cggcactgac gcgctgatgc tcggcgaagc caagcccagc
     6601 gcagatgcgg tcctcaatcg cgcgcaacgt ctccgcgtcg ggcttctctc ccgcgcggaa
     6661 gctaaccagc aggtgatagg tcttgtcggc ctcggaacgg gtgttgccgt gctgggtcgc
     6721 catcacctcg gccatgacag cgggcagggt gtttgcctcg cagttcgtga cgcgcacgtg
     6781 acccaggcgc tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc
     6841 gctcttcttg atggagcgca tggggacgtg cttggcaatc acgcgcaccc cccggccgtt
     6901 ttagcggcta aaaaagtcat ggctctgccc tcgggcggac cacgcccatc atgaccttgc
     6961 caagctcgtc ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga
     7021 accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca
     7081 ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt
     7141 tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc gccgccgcct
     7201 tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt gggctgccct
     7261 tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag
     7321 ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
     7381 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
     7441 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
     7501 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
     7561 aaaagcgctg cttccctgct gttttgtgga atatctaccg actggaaaca ggcaaatgca
     7621 ggaaattact gaactgaggg gacaggcgag agacgatgcc aaagagctac accgacgagc
     7681 tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg gcgtgatgag gctgcggttg
     7741 cgttcctggc ggtgagggcg gatgtcgagg cggcgttagc gtccggctat gcgctcgtca
     7801 ccatttggga gcacatgcgg gaaacgggga aggtcaagtt ctcctacgag acgttccgct
     7861 cgcacgccag gcggcacatc aaggccaagc ccgccgatgt gcccgcaccg caggccaagg
     7921 ctgcggaacc cgcgccggca cccaagacgc cggagccacg gcggccgaag caggggggca
     7981 aggctgaaaa gccggccccc gctgcggccc cgaccggctt caccttcaac ccaacaccgg
     8041 acaaaaagga tctactgtaa tggcgaaaat tcacatggtt ttgcagggca agggcggggt
     8101 cggcaagtcg gccatcgccg cgatcattgc gcagtacaag atggacaagg ggcagacacc
     8161 cttgtgcatc gacaccgacc cggtgaacgc gacgttcgag ggctacaagg ccctgaacgt
     8221 ccgccggctg aacatcatgg ccggcgacga aattaactcg cgcaacttcg acaccctggt
     8281 cgagctgatt gcgccgacca aggatgacgt ggtgatcctc tacgccggac gcatcgtggc
     8341 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     8401 ggaagatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     8461 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     8521 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
     8581 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     8641 ggcgtgcaag attccgaata ccgcaagcga caggccatgt ttgacagctt atcatcgata
//
