LOCUS       sequence-495374-        7462 bp    DNA     circular SYN 16-DEC-2025
DEFINITION  sfYFP fluorescent reporter for synthetic Mesorhizobium nifH 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  TITLE     Fluorescent reporters for rhizobium-legume symbiosis 
            [BIORXIV/2025/681774]
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7462)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pNDMS237
FEATURES             Location/Qualifiers
     source          1..7462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            140..248
                     /note="incP origin of transfer"
     CDS             complement(259..921)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      922..1693
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(4202..4288)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             4609..5322
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5404..5498
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5507..6157)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6263..7462
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 actacttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
       61 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
      121 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
      181 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
      241 gctggccgta ggccggccct accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg
      301 ctcgccatcg tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg
      361 ttcccattcc tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg
      421 ctggctgggc ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt
      481 cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac
      541 caccacggcg gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac
      601 gcggtcattg accacgtagg ccgacacggt gccggggccg ttgagcttca cgacggagat
      661 ccagcgctcg gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat
      721 gagcttggaa agagtctttt ggctgaccac cacggcgttc tggtggccca tctgcgcgac
      781 gaggtgatgc agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc
      841 atgcgctttg cgttccgttt ggacccagtg accgggcttg ttcttggctt gaatgccgat
      901 ttctctggac tgcgtggcca tgcttatctc catgcggtag gggtgccgca cggttgcggc
      961 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa
     1021 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc
     1081 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg
     1141 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct
     1201 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc
     1261 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg
     1321 aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg cccccactcg cataggcttg
     1381 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg
     1441 ccttcctctt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc
     1501 cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt
     1561 gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca
     1621 ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccgagc gaagcgagcc
     1681 agccggtggc cgctcgcggc catcgtccac atatccacgg gctggcaagg gagcgcagcg
     1741 accgcgcagg gcgaagcccg gagagcaagc ccgtaggggg ttactgcatg agcttgtgga
     1801 agtgtgctga gcataaatga ttctcctagc tgttctttgg gtaagtacgc catcaggacg
     1861 ttgtgagtgg cgcgattttt agcggctgaa atcagccctt gagcctgtcg gcaagtcgcg
     1921 tcatgaggtc catgcgctca tgcaggatcg ccacgaccaa cgcgggttcg cccgcacgcg
     1981 gcaggcaaaa aacgtagtgg tgttcgcagc gggccatccg cagcgcggga aagagttcgc
     2041 tcatgtcctt aaacgggcct tcgccggcgg caagcctggc tatgccctgt tccagcttag
     2101 cgatatagcg gcgcacctgc gccgcgcccc actcccggcg cgtgtagcgg atgatgccgc
     2161 gtagatcggc ttcggcctca gccgtgagga tgtaggccgt caagcgcgat ccccgctgag
     2221 ttcttcatca agaatttcgc cgacgctctt ggtggacacc ttgccggcaa gcccatcgtt
     2281 gatgcggttc cccagcatgg ttttcagttc ctgccatgcc tgatcggcat cagcgtcacc
     2341 ggggaacaga cgttcgaggg cgtattgctt aatggtcttg ccctgcaagg cggccagggc
     2401 tttcaggctc tggtgctgct ggtccgtcat gtcgattgtc aggcggctca ttggataacc
     2461 tccataaaat acacgtaacc acattagcac atatgtgggc gtgaggctac agcgcgaggc
     2521 gcattaaggt cgggaaaatg cgctaggcgc atttaaattg cgtattgctg taatgcgcca
     2581 tgccggctag actaggccca aatgggtata cccaatttga ccaaggggga cgcgatgagg
     2641 gcggccaagc actaccgaca acttctatcc atcgacttca acatcgaggc gctggccttc
     2701 gtgcctggac ccgacggcac acgcggccgg cgcatccacg tcctggggcg cgaggtccgc
     2761 gaccggcccg gcctggtcga gtacctttcg ccggcgttcg gctcgcgggt ggcgctggac
     2821 ggctactgca aggccaattt cgatgcagtg ctgcacctgg cgtaccccga tcatcagcaa
     2881 tggggccacg catgaagcgc cgaagctacg ccatgctgcg cgccgctgcc gcgctggccg
     2941 tcctggtcgt tgcctcgccg gcatgggccg agctgcgcgg cgaggtcgtg cgcatcatcg
     3001 acggcgacac catcgacgtg ctggtagaca agcagccggt gcgcgtgcgc ctggtggaca
     3061 ttgacgcgcc ggaaaagcgg caagccttcg gcgaacgtgc gcgccaggcg ctggccggca
     3121 tggtgttccg ccggcacgtc ctggtcgacg agaaggacac cgaccgttac ggccgcacgc
     3181 tgggcaccgt ctgggtcaac atggagctgg ccagccggcc gccgcagccg cgcaacgtca
     3241 acgccgcgat ggttcaccag ggcatggcgt gggcctatcg cttccacggc cgcgcggccg
     3301 accctgaaat gctgcggctc gaacaggagg cgcgaggcaa gcgcgtcggc ctctggtccg
     3361 atccgcacgc cgtcgagccg tggaaatggc gacgcgagag caacaaccgg agggacgaag
     3421 gttgaaggtc gcccgcatct acctgcgcgc cagtacggac gagcagaatc ttgaacgcca
     3481 ggagagcctt gtagcggcca cgcgggccgc cgggtactac gtcgccggca tctaccgcga
     3541 gaaggcgtcc ggcgcacgcg ccgaccggcc cgagctgctg cgcatgatcg cggacctgca
     3601 acctggtgaa gtcgtcgttg cggagaagat cgaccgcatc agccgcttgc cgttggccga
     3661 ggccgagcgc ctggttgcgt cgatccgggc caaaggggcc aagctggccg tgcctggcgt
     3721 ggtggacctg tcggagctgg ccgccgaggc gaacggagtg gcgaaaatcg ttctggaatc
     3781 cgtccaggac atgcttttga agctcgcctt gcagatggcc cgcgacgact acgaggatcg
     3841 gcgcgagcgt caacgtcagg gtgtccagtt ggcgaaggcc gccggccgct acaccggccg
     3901 caaacgtgac gccggcatgc acgaccgcat catcacgctt cgctccggcg gatcgagcat
     3961 tgccaagacg gccaagctgg tcggatgcag cccgagccag gtcaaacgag tgtgggcggc
     4021 ctggaacgcg cagcagcaaa aataaagccg ggcagtgccc ggcttttctc accttttcgc
     4081 gtcccgcagg gccgctgcga gcgccctacc tagatcctcg ctttccccct cggtgtagtc
     4141 cggccagggc acgaagggcg cggatgcgaa cctgttgagc aggtcagatc tagggcggcg
     4201 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     4261 ctttcgactg agcctttcgt tttatttgat gcctgccgaa ttcggatccg gagcgcagcc
     4321 aaaaacctgg tctgacttag gaccatccgc ttgagctcga ctctcgctgg atggcacccg
     4381 acgttttgtg tcggctttgt cgggtccgct acacaggctt ttatgggccc gatcacttgt
     4441 tttgccacaa gctaactagc tggaaagtaa tgaaaaaatt tccttttggt atcggccggt
     4501 caagttggca cgagtcttga gatccttggt gcgaggcggc aagagctgcc catcggcatt
     4561 tatgtcgcgg gcagccgcga tgataagaac aaggaaggaa aaactataat gcgtaaaggc
     4621 gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga tgtcaacggt
     4681 cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg
     4741 aagttcatct gtactactgg taaactgccg gtaccttggc cgactctggt aacgacgctg
     4801 acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca tgacttcttc
     4861 aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa ggatgacggc
     4921 acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa ccgcattgag
     4981 ctgaaaggca ttgactttaa agaagacggc aatatcctgg gccataagct ggaatacaat
     5041 tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat taaagcgaat
     5101 tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca ctaccagcaa
     5161 aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct gagctaccaa
     5221 agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct ggagttcgta
     5281 accgcagcgg gcatcacgca tggtatggat gaactgtaca aataatgaag gtctaacgca
     5341 tgagaaagcc cccggaagat caccttccgg gggctttttt attgcgccct tgagacgctc
     5401 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     5461 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggcaatcaa tcgtcaccct
     5521 ttctcggtcc ttcaacgttc ctgacaacga gcctcctttt cgccaatcca tcgacaatca
     5581 ccgcgagtcc ctgctcgaac gctgcgtccg gaccggcttc gtcgaaggcg tctatcgcgg
     5641 cccgcaacag cggcgagagc ggagcctgtt caacggtgcc gccgcgctcg ccggcatcgc
     5701 tgtcgccggc ctgctcctca agcacggccc caacagtgaa gtagctgatt gtcatcagcg
     5761 cattgacggc gtccccggcc gaaaaacccg cctcgcagag gaagcgaagc tgcgcgtcgg
     5821 ccgtttccat ctgcggtgcg cccggtcgcg tgccggcatg gatgcgcgcg ccatcgcggt
     5881 aggcgagcag cgcctgcctg aagctgcggg cattcccgat cagaaatgag cgccagtcgt
     5941 cgtcggctct cggcaccgaa tgcgtatgat tctccgccag catggcttcg gccagtgcgt
     6001 cgagcagcgc ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga acccccaacc
     6061 gttccgccag tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac aggtccaggg
     6121 cggcacggat cactgtattc ggctgcaact ttgtcatgct tgacacttta tcactgataa
     6181 acataatatg tccaccaact tatcagtgat aaagaatccg cgcgttcaat cggaccagcg
     6241 gaggctggtc cggaggccag acgtgaaacc caacataccc ctgatcgtaa ttctgagcac
     6301 tgtcgcgctc gacgctgtcg gcatcggcct gattatgccg gtgctgccgg gcctcctgcg
     6361 cgatctggtt cactcgaacg acgtcaccgc ccactatggc attctgctgg cgctgtatgc
     6421 gttggtgcaa tttgcctgcg cacctgtgct gggcgcgctg tcggatcgtt tcgggcggcg
     6481 gccaatcttg ctcgtgtcgc tggccggcgc cactgtcgac tacgccatca tggcgacagc
     6541 gcctttcctt tgggttctct atatcgggcg gatcgtggcc ggcatcaccg gggcgactgg
     6601 ggcggtagcc ggcgcttata ttgccgatat cactgatggc gatgagcgcg cgcggcactt
     6661 cggcttcatg agcgcctgtt tcgggttcgg gatggtcgcg ggacctgtgc tcggtgggct
     6721 gatgggcggt ttctcccccc acgctccgtt cttcgccgcg gcagccttga acggcctcaa
     6781 tttcctgacg ggctgtttcc ttttgccgga gtcgcacaaa ggcgaacgcc ggccgttacg
     6841 ccgggaggct ctcaacccgc tcgcttcgtt ccggtgggcc cggggcatga ccgtcgtcgc
     6901 cgccctgatg gcggtgttct tcatcatgca acttgtcgga caggtgccgg ccgcgctttg
     6961 ggtcattttc ggcgaggatc gctttcactg ggacgcgacc acgatcggca tttcgcttgc
     7021 cgcatttggc attctgcatt cactcgccca ggcaatgatc accggccctg tagccgcccg
     7081 gctcggcgaa aggcgggcac tcatgctcgg aatgattgcc gacggcacag gctacatcct
     7141 gcttgccttc gcgacacggg gatggatggc gttcccgatc atggtcctgc ttgcttcggg
     7201 tggcatcgga atgccggcgc tgcaagcaat gttgtccagg caggtggatg aggaacgtca
     7261 ggggcagctg caaggctcac tggcggcgct caccagcctg acctcgatcg tcggacccct
     7321 cctcttcacg gcgatctatg cggcttctat aacaacgtgg aacgggtggg catggattgc
     7381 aggcgctgcc ctctacttgc tctgcctgcc ggcgctgcgt cgcgggcttt ggagcggcgc
     7441 agggcaacga gccgatcgct ga
//
