LOCUS       sequence-115547-        8798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Paph-mCherry integration downstream of B. diazoefficiens
            (B. japonicum) scoI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8798)
  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 8798)
  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-mChe
FEATURES             Location/Qualifiers
     source          1..8798
                     /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     4558..4574
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4559..4575
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4590..5297)
                     /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(4865..4884)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5134..5152
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(7015..7386)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7419..7528
                     /note="incP origin of transfer"
     promoter        join(8779..8784,8785..8798,1..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 cggtaccggg ccccccctcg
     4561 aggtcgacgg tatcgataag cttcaattcc ttgtacagct cgtccatgcc gccggtggag
     4621 tggcggccct cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg
     4681 atgtccaact tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg
     4741 taggtggtct tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg
     4801 atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc
     4861 atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc
     4921 ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg
     4981 tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag
     5041 tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac
     5101 aggatgtccc aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg
     5161 gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg
     5221 gagccctcca tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc
     5281 tcgcccttgc tcaccataat tatatctcct tcttatttct agtcaagatc taagcttgat
     5341 atcgaattcc tgcagcccgg gggatccact agtatcctgt ctcttgatca gatcttgatc
     5401 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     5461 caaccttacc agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg
     5521 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     5581 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     5641 gcggactggc tttctacgtg ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc
     5701 ggcagcgtgc gagctcgaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga
     5761 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     5821 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     5881 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     5941 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     6001 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     6061 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     6121 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     6181 ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg
     6241 cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc
     6301 gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt gaccgtgtgc
     6361 ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat aattgacgat
     6421 atgatccagc cgaccaggct ttccacgccc gcgtgccgct ccatgtcgtt cgcgcggttc
     6481 tcggaaacgc gctgccgcgt ttcgtgattg tcacgctcaa gcccgtagtc ccgttcgagc
     6541 gtcgcgcaga ggtcagcgag ggcgcggtag gcccgatacg gctcatggat ggtgtttcgg
     6601 gtcgggtgaa tcttgttgat ggcgatatgg atgtgcaggt tgtcggtgtc gtgatgcacg
     6661 gcactgacgc gctgatgctc ggcgaagcca agcccagcgc agatgcggtc ctcaatcgcg
     6721 cgcaacgtct ccgcgtcggg cttctctccc gcgcggaagc taaccagcag gtgataggtc
     6781 ttgtcggcct cggaacgggt gttgccgtgc tgggtcgcca tcacctcggc catgacagcg
     6841 ggcagggtgt ttgcctcgca gttcgtgacg cgcacgtgac ccaggcgctc ggtcttgcct
     6901 tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg
     6961 gggacgtgct tggcaatcac gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg
     7021 ctctgccctc gggcggacca cgcccatcat gaccttgcca agctcgtcct gcttctcttc
     7081 gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc
     7141 ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag
     7201 ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag
     7261 caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc
     7321 gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc
     7381 agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg
     7441 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
     7501 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
     7561 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
     7621 aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct tccctgctgt
     7681 tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga actgagggga
     7741 caggcgagag acgatgccaa agagctacac cgacgagctg gccgagtggg ttgaatcccg
     7801 cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg tgagggcgga
     7861 tgtcgaggcg gcgttagcgt ccggctatgc gctcgtcacc atttgggagc acatgcggga
     7921 aacggggaag gtcaagttct cctacgagac gttccgctcg cacgccaggc ggcacatcaa
     7981 ggccaagccc gccgatgtgc ccgcaccgca ggccaaggct gcggaacccg cgccggcacc
     8041 caagacgccg gagccacggc ggccgaagca ggggggcaag gctgaaaagc cggcccccgc
     8101 tgcggccccg accggcttca ccttcaaccc aacaccggac aaaaaggatc tactgtaatg
     8161 gcgaaaattc acatggtttt gcagggcaag ggcggggtcg gcaagtcggc catcgccgcg
     8221 atcattgcgc agtacaagat ggacaagggg cagacaccct tgtgcatcga caccgacccg
     8281 gtgaacgcga cgttcgaggg ctacaaggcc ctgaacgtcc gccggctgaa catcatggcc
     8341 ggcgacgaaa ttaactcgcg caacttcgac accctggtcg agctgattgc gccgaccaag
     8401 gatgacgtgg tgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     8461 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatccttg aagctgtccc
     8521 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     8581 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
     8641 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     8701 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     8761 gcaagcgaca ggccatgttt gacagcttat catcgata
//
