LOCUS       sequence-501152-        6058 bp    DNA     circular SYN 25-FEB-2026
DEFINITION  Double-stranded RNA biomolecular fluorescence complementation sensor
            for bacteria, induced by arabinose..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  TITLE     Genetic biosensor for optimizing double-stranded RNA production by 
            bacterial symbionts
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6058)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 25, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBTK1211 (pDS-BiFC7)
FEATURES             Location/Qualifiers
     source          1..6058
                     /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(1812..2627)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1812..2627)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2535..2554
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2619..2638
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(2855..2949)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3147..4025)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4052..4336
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4259..4278
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4363..4385
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(4636..4638,4639..4641,4642..5100)
                     /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(4681..4702)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4942..4961)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             5395..5646
                     /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     5583..5604
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5858..5944
                     /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 cgggggcctt tctgcgttta taccgagcgg ccgcgtgtta caaccaatta
     1801 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1861 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1921 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1981 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     2041 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     2101 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2161 attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa
     2221 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     2281 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     2341 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     2401 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     2461 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     2521 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2581 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     2641 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     2701 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     2761 gagttgaagg atcagtcatg accaaaatcc cttaacaaaa gcccgctcat taggcgggct
     2821 actaatttaa atcgtaatta ttggggaccc ctggattctc accaataaaa aacgcccggc
     2881 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     2941 acaggagtcc aaggttctct tgcaggtggc acctgcttcg catctagact gatgagacgt
     3001 ggtagagcca caaacagccg gtacaagcaa cgatctccag gaccatctga atcatgcgcg
     3061 gatgacacga actcacgacg gcgatcacag acattaaccc acagtacaga cactgcgaca
     3121 acgtggcaat tcgtcgcaat acaacgttat gacaacttga cggctacatc attcactttt
     3181 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     3241 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     3301 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     3361 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     3421 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     3481 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     3541 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     3601 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     3661 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     3721 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     3781 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     3841 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     3901 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     3961 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     4021 gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga
     4081 cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt
     4141 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     4201 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
     4261 gccatagcat ttttatccat aagattagcg gatactacct gacgcttttt atcgcaactc
     4321 tctactgttt ctccataccc gtttttttgg gctagaaata attttgttta actttaagaa
     4381 ggagatatac atatgcgggg ttctatgccc tccaaacttg cccttatcca agaacttccc
     4441 gatcggattc aaaccgctgt tgaagcagct atgggcatgt cgtatcaagc cgcgcccaac
     4501 aatgtgcgtc gggaccttga caacctgcac gcctgcctta acaaggccaa acttactgtc
     4561 tcgcgtatgg tcacgagcct gcttgaaaag cctagcgtgg tagcatacct ggagggcaaa
     4621 gcgcccggca gatctatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4681 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4741 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     4801 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     4861 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4921 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4981 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     5041 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tctcaccgcc
     5101 tgattgtatt gcgacgaatt gccattctat acagaatact agagaaagag gggaaatact
     5161 agtatgccaa gcaaactcgc gctaatccag gaacttcccg accgcattca aacggcggtg
     5221 gaagcagcca tgggaatgag ctaccaagcc gcaccgaaca acgtgcgcag ggacctcgac
     5281 aacctgcacg cttgcctaaa caaggcaaaa ctaacggtaa gtcggatggt aacatcactg
     5341 ctggagaaac ccagcgtggt ggcataccta gagggaaagg ccccccgagg atccgacaag
     5401 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     5461 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5521 gacaaccact acctgagcta ccagtccaaa ctgagcaaag accccaacga gaagcgcgat
     5581 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5641 tacaagtaaa tccgctggaa atctgctcgt cagtggtgct cacactgacg aatcatgtac
     5701 agatcatacc gatgactgcc tggcgactca caactaagca agacagccgg aaccagcgcc
     5761 ggcgaacacc actgcatata tggcatatca caacagtcca cgtctcaagc agttacagag
     5821 atgttacgaa cgcgcatcac ctgcaaggcg aaggcatcaa ataaaacgaa aggctcagtc
     5881 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     5941 aaatccgccg ccctagacag ctgggcgcgc cccccctacg ggcttgctct ccgggcttcg
     6001 ccctgcgcgg tcgctgcgct cccttgccag cccgtggata tgtggacgat ggccgcga
//
