LOCUS       sequence-500785-        5983 bp    DNA     circular SYN 23-FEB-2026
DEFINITION  Double-stranded RNA biomolecular fluorescence complementation sensor
            for bacteria, induced by arabinose. Variant of pDS-BiFC7 with AmpR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5983)
  TITLE     Genetic biosensor for optimizing double-stranded RNA production by 
            bacterial symbionts
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5983)
  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            "Unpublished"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: pBTK1212
FEATURES             Location/Qualifiers
     source          1..5983
                     /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"
     CDS             complement(3072..3950)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3977..4261
                     /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     4184..4203
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4288..4310
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(4561..4563,4564..4566,4567..5025)
                     /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(4606..4627)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4867..4886)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             5320..5571
                     /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     5508..5529
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5783..5869
                     /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 gttatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
     3121 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga
     3181 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
     3241 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
     3301 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
     3361 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
     3421 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
     3481 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
     3541 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
     3601 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
     3661 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     3721 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     3781 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     3841 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     3901 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     3961 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
     4021 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
     4081 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
     4141 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
     4201 tccataagat tagcggatac tacctgacgc tttttatcgc aactctctac tgtttctcca
     4261 tacccgtttt tttgggctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     4321 cggggttcta tgccctccaa acttgccctt atccaagaac ttcccgatcg gattcaaacc
     4381 gctgttgaag cagctatggg catgtcgtat caagccgcgc ccaacaatgt gcgtcgggac
     4441 cttgacaacc tgcacgcctg ccttaacaag gccaaactta ctgtctcgcg tatggtcacg
     4501 agcctgcttg aaaagcctag cgtggtagca tacctggagg gcaaagcgcc cggcagatct
     4561 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4621 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4681 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4741 ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     4801 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4861 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4921 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4981 aagctggagt acaactacaa cagccacaac gtctatctca ccgcctgatt gtattgcgac
     5041 gaattgccat tctatacaga atactagaga aagaggggaa atactagtat gccaagcaaa
     5101 ctcgcgctaa tccaggaact tcccgaccgc attcaaacgg cggtggaagc agccatggga
     5161 atgagctacc aagccgcacc gaacaacgtg cgcagggacc tcgacaacct gcacgcttgc
     5221 ctaaacaagg caaaactaac ggtaagtcgg atggtaacat cactgctgga gaaacccagc
     5281 gtggtggcat acctagaggg aaaggccccc cgaggatccg acaagcagaa gaacggcatc
     5341 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     5401 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     5461 agctaccagt ccaaactgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     5521 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaatccgc
     5581 tggaaatctg ctcgtcagtg gtgctcacac tgacgaatca tgtacagatc ataccgatga
     5641 ctgcctggcg actcacaact aagcaagaca gccggaacca gcgccggcga acaccactgc
     5701 atatatggca tatcacaaca gtccacgtct caagcagtta cagagatgtt acgaacgcgc
     5761 atcacctgca aggcgaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     5821 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
     5881 gacagctggg cgcgcccccc ctacgggctt gctctccggg cttcgccctg cgcggtcgct
     5941 gcgctccctt gccagcccgt ggatatgtgg acgatggccg cga
//
