LOCUS       sequence-500787-        5844 bp    DNA     circular SYN 23-FEB-2026
DEFINITION  Double-stranded RNA biomolecular fluorescence complementation sensor
            for bacteria, induced by vanillic acid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5844)
  TITLE     Genetic biosensor for optimizing double-stranded RNA production by 
            bacterial symbionts
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5844)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 23, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBTK1210 (pDS-BiFC6)
FEATURES             Location/Qualifiers
     source          1..5844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..772
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             773..1435
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     oriT            1446..1554
                     /note="incP origin of transfer"
     terminator      complement(1712..1751)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(1755..2546,2547..2615))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2616..2707)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2780..2874)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_RNA        3157..3206
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(3488..3490,3491..3493,3494..3952)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(3533..3554)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3794..3813)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4247..4498
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     4435..4456
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5644..5730
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
       61 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
      121 cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt
      181 aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac
      241 accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc
      301 gcctggaacg acccaagcct atgcgagtgg gggcagtcga agggcgaagc ccgcccgcct
      361 gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc
      421 aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg
      481 ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga
      541 actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac
      601 gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt
      661 aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct
      721 gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac
      781 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
      841 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
      901 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
      961 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     1021 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     1081 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     1141 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     1201 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     1261 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     1321 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     1381 ggacgatggc gagccgttgg agccgccgac acgggtaacg ctgccgcgcc ggtagggccg
     1441 gcctacggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     1501 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     1561 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     1621 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     1681 gcggaaaagg acaacgcgcg gaccggggtc gtccaaaaaa aaaggctcca aaaggagcct
     1741 ttaattgtat cggtttacca atgtttaatc aggctcgcgc caatttccgc aatctgacgg
     1801 ttacgttcat ccatcgtcgc ctggctgccg gtggtataaa tcaccacaat acggctcggt
     1861 ttaccatccg ggcccagcgc cgcaataatg ccacggctgc cacgttcgcc cgcaccgctt
     1921 ttatccgcaa taaaccagcc agccggcagc gcgctacgca gcagcggacc cgccacttta
     1981 tccgcttcca tccaatcaat cagttgctgg cggcttgcca gggtcagcag ctcacccgtc
     2041 agcagtttac gcagggtggt cgccattgct gccggcatgg tggtatcacg ttcatcgttc
     2101 ggaatcgctt cgttcagttc cggttcccaa cgatccagac gggtcacatg atcgcccatg
     2161 ttatgcagaa acgcggtcag ttctttcgga ccgccaatgg tcgtcagcag caggttcgcc
     2221 gcggtgttat cgctcatggt aatcgccgcg ctgcacagtt cacgcacggt catgccatcg
     2281 gtcagatgtt tttcggtcac cgggctatat tccaccagat cgttctggct ataatgaata
     2341 cgacggccca gttgttcctg gcccgcatcc acacggctca gaaccgcacc gcacagcagc
     2401 actttaaagg tgctcatcat cggaaaacgt tcttccggac gaaaagattc cagaattttg
     2461 ccgctgttca gatccagttc aatatagccc acgcgcgcac ccagttgatc ttccgcatct
     2521 ttcactttca ccagggtttc cggatgcgca aacaccggca ggcaaaacgc cgcaaaaaac
     2581 ggaatcagcg ccacacgaaa atgctgaatg ctcatactct tcctttttca atattattga
     2641 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2701 aaacaaatag gggaattaaa aaaaagcccg ctcattaggc gggctactaa tttaaatcgt
     2761 aattattggg gacccctgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     2821 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaaggt
     2881 tctcttgcag gtggcacctg cttcgcatct agactgatga gacgtggtag agccacaaac
     2941 agccggtaca agcaacgatc tccaggacca tctgaatcat gcgcggatga cacgaactca
     3001 cgacggcgat cacagacatt aacccacagt acagacactg cgacaacgtg gcaattcgtc
     3061 gcaatacaac ggtttctggt atcccgctta acgatcgttg gctgattgga tccaattgac
     3121 agctagctca gtcctaggta ccattggatc caatagctgt caccggatgt gctttccggt
     3181 ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaat actagagaaa
     3241 gaggggaaat actagtatgc cctccaaact tgcccttatc caagaacttc ccgatcggat
     3301 tcaaaccgct gttgaagcag ctatgggcat gtcgtatcaa gccgcgccca acaatgtgcg
     3361 tcgggacctt gacaacctgc acgcctgcct taacaaggcc aaacttactg tctcgcgtat
     3421 ggtcacgagc ctgcttgaaa agcctagcgt ggtagcatac ctggagggca aagcgcccgg
     3481 cagatctatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3541 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3601 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
     3661 gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca
     3721 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3781 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3841 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3901 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatctcaccg cctgattgta
     3961 ttgcgacgaa ttgccattct atacagaata ctagagaaag aggggaaata ctagtatgcc
     4021 aagcaaactc gcgctaatcc aggaacttcc cgaccgcatt caaacggcgg tggaagcagc
     4081 catgggaatg agctaccaag ccgcaccgaa caacgtgcgc agggacctcg acaacctgca
     4141 cgcttgccta aacaaggcaa aactaacggt aagtcggatg gtaacatcac tgctggagaa
     4201 acccagcgtg gtggcatacc tagagggaaa ggccccccga ggatccgaca agcagaagaa
     4261 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     4321 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4381 ctacctgagc taccagtcca aactgagcaa agaccccaac gagaagcgcg atcacatggt
     4441 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     4501 aatccctcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct tttttcgttt
     4561 tggtccaatg gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
     4621 gcccggaaga gagtcaattc aggggggtga ataatggaca tgcctcgtat taaaccgggt
     4681 cagcgtgtta tgatggcact gcgtaaaatg attgcaagcg gtgaaatcaa aagtggtgaa
     4741 cgtattgcag aaattccgac cgcagcagca ctgggtgtta gccgtatgcc ggttcgtatc
     4801 gcactgcgtt cactggaaca agaaggtctg gttgttcgtc tgggtgcacg tggttatgca
     4861 gcccgtggtg ttagcagcga tcagattcgt gatgcaattg aagttcgtgg tgttctggaa
     4921 ggttttgcag cacgtcgtct ggcagaacgt ggtatgaccg cagaaaccca tgcacgtttt
     4981 gttgtactga ttgcagaagg tgaagcactg tttgcagccg gtcgcctgaa tggtgaagat
     5041 ctggatcgtt atgccgcata taatcaggca tttcatgata ccctggttag cgcagcaggt
     5101 aatggtgcag ttgaaagcgc actggcacgt aatggttttg aaccgtttgc agcagccggt
     5161 gcactggccc tggatctgat ggacctgtct gccgaatatg aacatctgct ggcagcacat
     5221 cgtcagcatc aggcagttct ggatgcagtt agctgtggtg atgccgaagg tgcagaacgt
     5281 attatgcgtg atcatgcact ggcagcaatt cgtaatgcaa aagtttttga agcagcagca
     5341 agcgcaggcg caccgctggg tgcagcatgg tcaattcgtg cagattgata aggatcctaa
     5401 ttggtaacga atcagacaat tgacggctcg agggagtagg ctggaaatct gctcgtcagt
     5461 ggtgctcaca ctgacgaatc atgtacagat cataccgatg actgcctggc gactcacaac
     5521 taagcaagac agccggaacc agcgccggcg aacaccactg catatatggc atatcacaac
     5581 agtccacgtc tcaagcagtt acagagatgt tacgaacgcg catcacctgc aaggcgaagg
     5641 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     5701 tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacagctgg gcgcgccccc
     5761 cctacgggct tgctctccgg gcttcgccct gcgcggtcgc tgcgctccct tgccagcccg
     5821 tggatatgtg gacgatggcc gcga
//
