LOCUS       sequence-495377-        7385 bp    DNA     circular SYN 16-DEC-2025
DEFINITION  sfCFP fluorescent reporter for synthetic Rhizobium nifH promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  TITLE     Fluorescent reporters for rhizobium-legume symbiosis 
            [BIORXIV/2025/681774]
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7385)
  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            "Unpublished"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: pNDMS291
FEATURES             Location/Qualifiers
     source          1..7385
                     /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             4535..5248
                     /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
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5327..5421
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5430..6080)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6186..7385
                     /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 gagctcgaca
     4321 cgacattgtc gtcacctttg tcgggtccgc gacaagcttt ttgttcctcg atcccgtatg
     4381 ttttcgccta aatatctgaa acagcagcaa attatttttc gttagctcaa tcggcccact
     4441 tggcacgagt cttgaaattt attgcgaggc ggcggagctg ccggccgcat ccgtgcggca
     4501 taaccacggt tgtgaacaaa tgaaggaagg ccaaatgcgt aaaggcgaag agctgttcac
     4561 tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt
     4621 gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac
     4681 tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt ggggtgttca
     4741 gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc
     4801 ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg
     4861 tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga
     4921 ctttaaagaa gacggcaata tcctgggcca taagctggaa tacaattaca tcagcgacaa
     4981 tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca
     5041 caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg
     5101 tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa
     5161 agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat
     5221 cacgcatggt atggatgaac tgtacaaata aaggtctaac gcatgagaaa gcccccggaa
     5281 gatcaccttc cgggggcttt tttattgcgc ccttgagacg ctcttggact cctgttgata
     5341 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     5401 gcgtttttta ttggtgagaa tccaggcaat caatcgtcac cctttctcgg tccttcaacg
     5461 ttcctgacaa cgagcctcct tttcgccaat ccatcgacaa tcaccgcgag tccctgctcg
     5521 aacgctgcgt ccggaccggc ttcgtcgaag gcgtctatcg cggcccgcaa cagcggcgag
     5581 agcggagcct gttcaacggt gccgccgcgc tcgccggcat cgctgtcgcc ggcctgctcc
     5641 tcaagcacgg ccccaacagt gaagtagctg attgtcatca gcgcattgac ggcgtccccg
     5701 gccgaaaaac ccgcctcgca gaggaagcga agctgcgcgt cggccgtttc catctgcggt
     5761 gcgcccggtc gcgtgccggc atggatgcgc gcgccatcgc ggtaggcgag cagcgcctgc
     5821 ctgaagctgc gggcattccc gatcagaaat gagcgccagt cgtcgtcggc tctcggcacc
     5881 gaatgcgtat gattctccgc cagcatggct tcggccagtg cgtcgagcag cgcccgcttg
     5941 ttcctgaagt gccagtaaag cgccggctgc tgaaccccca accgttccgc cagtttgcgt
     6001 gtcgtcagac cgtctacgcc gacctcgttc aacaggtcca gggcggcacg gatcactgta
     6061 ttcggctgca actttgtcat gcttgacact ttatcactga taaacataat atgtccacca
     6121 acttatcagt gataaagaat ccgcgcgttc aatcggacca gcggaggctg gtccggaggc
     6181 cagacgtgaa acccaacata cccctgatcg taattctgag cactgtcgcg ctcgacgctg
     6241 tcggcatcgg cctgattatg ccggtgctgc cgggcctcct gcgcgatctg gttcactcga
     6301 acgacgtcac cgcccactat ggcattctgc tggcgctgta tgcgttggtg caatttgcct
     6361 gcgcacctgt gctgggcgcg ctgtcggatc gtttcgggcg gcggccaatc ttgctcgtgt
     6421 cgctggccgg cgccactgtc gactacgcca tcatggcgac agcgcctttc ctttgggttc
     6481 tctatatcgg gcggatcgtg gccggcatca ccggggcgac tggggcggta gccggcgctt
     6541 atattgccga tatcactgat ggcgatgagc gcgcgcggca cttcggcttc atgagcgcct
     6601 gtttcgggtt cgggatggtc gcgggacctg tgctcggtgg gctgatgggc ggtttctccc
     6661 cccacgctcc gttcttcgcc gcggcagcct tgaacggcct caatttcctg acgggctgtt
     6721 tccttttgcc ggagtcgcac aaaggcgaac gccggccgtt acgccgggag gctctcaacc
     6781 cgctcgcttc gttccggtgg gcccggggca tgaccgtcgt cgccgccctg atggcggtgt
     6841 tcttcatcat gcaacttgtc ggacaggtgc cggccgcgct ttgggtcatt ttcggcgagg
     6901 atcgctttca ctgggacgcg accacgatcg gcatttcgct tgccgcattt ggcattctgc
     6961 attcactcgc ccaggcaatg atcaccggcc ctgtagccgc ccggctcggc gaaaggcggg
     7021 cactcatgct cggaatgatt gccgacggca caggctacat cctgcttgcc ttcgcgacac
     7081 ggggatggat ggcgttcccg atcatggtcc tgcttgcttc gggtggcatc ggaatgccgg
     7141 cgctgcaagc aatgttgtcc aggcaggtgg atgaggaacg tcaggggcag ctgcaaggct
     7201 cactggcggc gctcaccagc ctgacctcga tcgtcggacc cctcctcttc acggcgatct
     7261 atgcggcttc tataacaacg tggaacgggt gggcatggat tgcaggcgct gccctctact
     7321 tgctctgcct gccggcgctg cgtcgcgggc tttggagcgg cgcagggcaa cgagccgatc
     7381 gctga
//
