LOCUS       sequence-496758-        7859 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  PBT0463 biosensor circuit in pNBU2 integrative Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7859)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 7859)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAH108
FEATURES             Location/Qualifiers
     source          1..7859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             207..893
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGT
                     VSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     complement(337..355)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      919..946
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             1262..1885
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1335..1354)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    2050..2068
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(2155..2157,2158..2160,2161..2874)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(2200..2221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      2895..2922
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4902..5693,5694..5762))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5525..5544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7054..7442
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7613..7722
                     /note="incP origin of transfer"
ORIGIN
        1 caattgggct accttttttt tgttttgttt gcaatggtta atctattgtt aaaatttaaa
       61 gtttcacttg aactttcaaa taatgttctt atatttgcag tgtcgaaaga aacaaagtag
      121 gactgatcac acaacattta aaaaataaca ttatgaaagc actcgaaaag agattcaaag
      181 gtaatattaa caataattta ttttcaatgg atagcaccga ggcagtgatc aaggagttca
      241 tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
      301 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      361 gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct
      421 tcatcaagca ccccgccgac atccccgact actggaagca gtccttcccc gagggcttca
      481 agtgggagcg cgtgatgatc ttcgaggacg gcggcaccgt gtccgtgacc caggacacct
      541 ccctggagga cggcaccctg atctacaagg tgaagctccg cggcggcaac ttccctcctg
      601 acggccccgt aatgcagaag aggacaatgg gctgggaagc atccaccgag cggttgtacc
      661 ccgaggacgt cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc
      721 gctacctggc ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg
      781 ccttcaacat cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg
      841 aacagtacga acgctccgtg gcccgccact ccaccggcgg ctccggtggc tcctaataaa
      901 tcctagctga ttagctcgag aaggccatcc tgacggatgg cctttttttt gtatggggac
      961 tctgagaacc ctacctttca aaccccgact cacgcaatac aagaagagct attacgtagt
     1021 tgttacgtta gctactatcg gcgtcatgct acatattccc aagtggtatt gcgaaaaaat
     1081 caactaatca taacattacg atcatctgct caacaccttg ttgtcggaca cttcatgttt
     1141 gacctttaga actgtttttt taatctcgga aggtaatcta tttatttaat aaaatattca
     1201 tttttttgag tggaaggata tataccctgc cactcaaatc ttgagtacta ataacaaatc
     1261 aatgtccaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt
     1321 cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac
     1381 attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt
     1441 agataggcac catactcact tttgcccttt agaaggggaa agctggcaag attttttacg
     1501 taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca
     1561 tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt
     1621 atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg tggggcattt
     1681 tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac
     1741 acctactact gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca
     1801 aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca
     1861 acttaaatgt gaaagtgggt cttaatggcc acatcggttc ctgctagaag gcccatcctg
     1921 acggatggcc ttttggtatt cgagcattac ctcatacttt ggatttttgt tttgtttgca
     1981 atggttaatc tattgttaaa atttaaagtt tcacttgaac tttcaaataa tgttcttata
     2041 tttgcagtgt ccctatcagt gatagagagt aggatcacac aacatttaaa aaataacatt
     2101 atgaaagcac tcgaaaagag attcaaaggt aatattaaca ataatttatt ttcaatggtg
     2161 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2221 gtaaacggcc acaagttcag cgtccgcggc gagggcgagg gcgatgccac caacggcaag
     2281 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2341 accaccttag gctacggcgt ggcctgcttc gcccgctacc ccgaccacat gaagcagcac
     2401 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctctttcaag
     2461 gacgacggta cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2521 cgcatcgtgc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2581 gagtacaact tcaacagcca caaggtctat atcacggccg acaagcagaa gaacggcatc
     2641 aaggctaact tcaagacccg ccacaacgtt gaggacggcg gcgtgcagct cgccgaccac
     2701 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2761 agccatcagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgaag
     2821 gagagggtga ccgccgccgg gattacacat gacatggacg agctgtacaa gtaatagcgt
     2881 tcagctcgac tgtgagaagg ccatcctgac ggatggcctt tttttttcgt tataactagt
     2941 tgcgttccta gccgctacta gtgtaattgc ctatcttcca gtgatggaac agcatttgtg
     3001 cattggctgc aacaatcagc cttacttgtg cctgttctat ttccgaaccg accgcttgta
     3061 tgaatgcatc aaaattcgtt ttctctacgt tggattcctt gttgctcata ttgtgatgat
     3121 aatttctaca aatatagtca ttggtaacta tctatgaaac tgtttgatac ttttatagtt
     3181 gattaaactt gttcatggca tttgccttaa tatcatccgc tatgtcaatg tagggtttca
     3241 tagctttgta gtcgctgtgt cccgtccatt tcatgaccac ctgtgccggg attccgagag
     3301 ccagcgcatt gcagatgaat gtcctttttc ctgcatgggt actgagcaaa gcgtatttgt
     3361 gtgtgacttc atcaatacgt tcatttccct tgtagtaggt ttcccgtaca ggctcgttga
     3421 tttctgccag ttcgcccagc tctttcaggt aatcgttcat cttctggttg ctgatgacgg
     3481 gcagagccat gtaattctcg aaatggatgt ccttgtattt gtccagtatg gctttgctgt
     3541 atttgttcag ttcaatcgtc aggctgtcgg cagtcttgac tgtggttatt tcgatgtggt
     3601 cggacttcac atcgcttctt ttcagattgc gaacatccga ataccgcaaa ctcgtaaagc
     3661 agcagaacag gaaaacatca cgcacacgtt ccaggtattg cttatccttg ggtatctggt
     3721 agtctttcag cttgttcagt tcatcccaag tcaggaagat tacttttttc gaggtggttt
     3781 tcagtttcgg tttgaacgta tcgtatgcaa tgttctgatg atgtcctttc ttgaagctcc
     3841 agcgcaggaa ccatttgagg aatcccattt gcttgccgat ggtgctgttt ctcatatcct
     3901 tggtgtcacg caggaagttg acgtattcgt tcaatccaaa ctcgttgaaa tagttgaacg
     3961 ttgcatcctc cttgaactct ttgaggtggt tcctcactgc tgcaaatttt tcataggtgg
     4021 atgccgtcca gttattctgg ttaccgcact cttttacaaa ctcatcgaac acctcccaaa
     4081 agctgacagg ggcttcttcc ggctgttctt cactggtgtc tttcattctc atgttgaaag
     4141 cttccttcaa ctgttgggtc gttggcatga cctcctgcac ctcaaattcc ttgaaaatat
     4201 tctggatttc ggcatagtat ttcagcaagt ccgtattgat ttcggctgca ctttgcttta
     4261 gcttgttggt acatccgttc tttacccgct gcttatctgc atcccatttg gctacgtcaa
     4321 tccggtagcc cgttgtaaac tcgatacgtt ggctggcaaa gatgacacgc atacggatgg
     4381 gtacgttctc tacgattggc acaccgttct ttttccggct ctccaatgca aaaatgatgt
     4441 tgcgcttgat attcataatt gggtgcgttt gaaattctac acccaaatat acacccaatt
     4501 attgagatag caaaagacat ttagaaacat ttacttttac tctatattgt aatttacact
     4561 tgattatcag tcgtttgcag ttttatgata ttctgtgaaa gtataagttc gagagcctgt
     4621 ctctccgcaa aaaacgctga aaatcagcag attgcaaaac aaacaccctg ttttacaccc
     4681 aagaatgtaa agtcgggtgt ttttgtttta tttaagataa tacaaccact acataataaa
     4741 agagtagcga tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt
     4801 ttgatttctc aggactttac atcgtcctga aagtatttgt tggtaccggt accgaggacg
     4861 cgacagctgc agacgcgtta tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5041 cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5761 atactcttcc tttttcaatc atggccgcgg gattaaaagt cggggattgg tgaacaaaaa
     5821 ggtgtttctc tctttaagag aaatatcgtt ttgctaaaca gttgatattg aggtatcatt
     5881 ttatcgtaaa agacattttt gctcaacaat tgcttgacgg aaatcaacaa attttagcat
     5941 tttgtaaaaa agtcgctata taatttggtg aattggagtt attttcatat ttttgcatcc
     6001 cgaagagttt ctcttaaaga gagaaacatc ttttgcatac cttttccgac cgaattttta
     6061 tgtcgtaaag aggggctttg cagggggtgg actcagaaag atgagaatag atgactattg
     6121 tagttgaaac acatagaaag ttgctgatat acagaccgat acgcatatcg ggatgaacca
     6181 tgagtacgtt cttttctcaa aaaacataaa tattcgaaaa gagatgcaat aaattaagga
     6241 gaggttataa tgaacaaagt aaatataaaa gatagtcaaa attttattac ttcaaaatat
     6301 cacatagaaa aaataatgaa ttgcataagt ttagatgaaa aagataacat ctttgaaata
     6361 ggtgcaggga aaggtcattt tactgctgga ttggtaaaga gatgtaattt tgtaacggcg
     6421 atagaaattg attctaaatt atgtgaggta actcgtaata agctcttaaa ttatcctaac
     6481 tatcaaatag taaatgatga tatactgaaa tttacatttc ctagccacaa tccatataaa
     6541 atatttggca acatacctta caacataagc acaaatataa ttcgaaaaat tgtttttgaa
     6601 agttcagcca caataagtta tttaatagtg gaatatggtt ttgctaaaat gttattagat
     6661 acaaacagat cactagcatt gctgttaatg gcagaggtag atatttctat attagcaaaa
     6721 attcctaggt attatttcca tccaaaacct aaagtggata gcacattaat tgtattaaaa
     6781 agaaagccag caaaaatggc atttaaagag agaaaaaaat atgaaacttt tgtaatgaaa
     6841 tgggttaaca aagagtacga aaaactgttt acaaaaaatc aatttaataa agctttaaaa
     6901 catgcgagaa tatatgatat aaacaatatt agtttcgaac aatttgtatc gctatttaat
     6961 agttataaaa tatttaacgg ctaaaaacaa taggccacat gcaactgtaa atgtttacgc
     7021 gggtaccgac accgcggtgg aggggaattc ccatgtcagc cgttaagtgt tcctgtgtca
     7081 ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg
     7141 tccacaaccg ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa
     7201 aacttaaaac cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca
     7261 tgagagctta gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt
     7321 tagccatgag ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta
     7381 gtacgtgaaa catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga
     7441 tcctctacgc cggacgcatc gtggccggat caattccgtt ttccgctgca taaccctgct
     7501 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     7561 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     7621 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     7681 tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag
     7741 atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca
     7801 cggaattgat ccccctcgaa ttggaattct agcctggaat agggaagcac cacatcacc
//
