LOCUS       sequence-496765-        8682 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  PBT0779+10xTetO biosensor cicuit in pNBU2 integrative Bacteroides 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8682)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 8682)
  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: pJB163
FEATURES             Location/Qualifiers
     source          1..8682
                     /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"
     protein_bind    1023..1041
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1070..1088
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1119..1137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1151..1169
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1193..1211
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1239..1257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1281..1299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1329..1347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1378..1396
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1420..1438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1473..1544
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1560..1587
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2085..2708
                     /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(2158..2177)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    2873..2891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(2978..2980,2981..2983,2984..3697)
                     /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(3023..3044)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      3718..3745
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(5725..6516,6517..6585))
                     /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     6348..6367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7877..8265
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            8436..8545
                     /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 aaccccgagc ggccgcttct agagatcgat tttagagctg
     1021 cctccctatc agtgatagag attgctacaa ctgcaaggta gtcccggtat ccctatcagt
     1081 gatagagact gatagctacg agtctgagat cctacttgtc cctatcagtg atagagacag
     1141 taccgagttt tccctatcag tgatagagag ctcatactgc gacagagtct tttccctatc
     1201 agtgatagag ataggctcct ttgtcaaaag tgatcgcttc cctatcagtg atagagactt
     1261 cacgggataa tgtgttctac tccctatcag tgatagagag cttaagtcaa tccaatgcgt
     1321 tcaggtgctc cctatcagtg atagagaaac tccgtcatcg tggatgtcaa tactgactcc
     1381 ctatcagtga tagagattga tttcaccgct ggcgtcaatt ccctatcagt gatagagact
     1441 taggtgtccc tagaattact agagccaggc atcaaataaa acgaaaggct cagtcgaaag
     1501 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     1561 gctcaccttc gggtgggcct ttctgcgttt atatatgatg gcagcatcca ttgcctgaaa
     1621 aataaccaga accgagtagc ggtcttgtgc ataaagcata ctttgcagtt ctgacagctt
     1681 ttcgatattt tgggctaact gttctttggc atcctgtttg gacatatcag cagtaaatga
     1741 gggattgaaa tcggatacat gatgttcctt ccccggtttg gctgtaagct cttttaagac
     1801 ttcttttttc attgttgtat aattaggttg ttataggtat ataacagttg aataaagaaa
     1861 tcagttcttg ccaaattaag agcgttgaat agtaattccc caattatgta tcaacagatt
     1921 ctacataatt ccacactaat tccacaatta tatttatatt gtctgctgtt tatatgtttg
     1981 attatcagtt atatatggcg gagtgaaacg tttggcatct gctttgctat atattaagta
     2041 caagaaacaa tagttctatt attcacactt aaaaaattac gattatgtcc agattagata
     2101 aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa
     2161 caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa
     2221 aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc
     2281 acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt
     2341 ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta
     2401 cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt
     2461 cactagagaa tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat
     2521 tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta
     2581 tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct
     2641 tcttattcgg ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg
     2701 ggtcttaatg gccacatcgg ttcctgctag aaggcccatc ctgacggatg gccttttggt
     2761 attcgagcat tacctcatac tttggatttt tgttttgttt gcaatggtta atctattgtt
     2821 aaaatttaaa gtttcacttg aactttcaaa taatgttctt atatttgcag tgtccctatc
     2881 agtgatagag agtaggatca cacaacattt aaaaaataac attatgaaag cactcgaaaa
     2941 gagattcaaa ggtaatatta acaataattt attttcaatg gtgagcaagg gcgaggagct
     3001 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3061 cagcgtccgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
     3121 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct taggctacgg
     3181 cgtggcctgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3241 catgcccgaa ggctacgtcc aggagcgcac catctctttc aaggacgacg gtacctacaa
     3301 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg tgctgaaggg
     3361 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag
     3421 ccacaaggtc tatatcacgg ccgacaagca gaagaacggc atcaaggcta acttcaagac
     3481 ccgccacaac gttgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     3541 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagccatc agtccaagct
     3601 gagcaaagac cccaacgaga agcgcgatca catggtcctg aaggagaggg tgaccgccgc
     3661 cgggattaca catgacatgg acgagctgta caagtaatag cgttcagctc gactgtgaga
     3721 aggccatcct gacggatggc cttttttttt cgttataact agttgcgttc ctagccgcta
     3781 ctagtgtaat tgcctatctt ccagtgatgg aacagcattt gtgcattggc tgcaacaatc
     3841 agccttactt gtgcctgttc tatttccgaa ccgaccgctt gtatgaatgc atcaaaattc
     3901 gttttctcta cgttggattc cttgttgctc atattgtgat gataatttct acaaatatag
     3961 tcattggtaa ctatctatga aactgtttga tacttttata gttgattaaa cttgttcatg
     4021 gcatttgcct taatatcatc cgctatgtca atgtagggtt tcatagcttt gtagtcgctg
     4081 tgtcccgtcc atttcatgac cacctgtgcc gggattccga gagccagcgc attgcagatg
     4141 aatgtccttt ttcctgcatg ggtactgagc aaagcgtatt tgtgtgtgac ttcatcaata
     4201 cgttcatttc ccttgtagta ggtttcccgt acaggctcgt tgatttctgc cagttcgccc
     4261 agctctttca ggtaatcgtt catcttctgg ttgctgatga cgggcagagc catgtaattc
     4321 tcgaaatgga tgtccttgta tttgtccagt atggctttgc tgtatttgtt cagttcaatc
     4381 gtcaggctgt cggcagtctt gactgtggtt atttcgatgt ggtcggactt cacatcgctt
     4441 cttttcagat tgcgaacatc cgaataccgc aaactcgtaa agcagcagaa caggaaaaca
     4501 tcacgcacac gttccaggta ttgcttatcc ttgggtatct ggtagtcttt cagcttgttc
     4561 agttcatccc aagtcaggaa gattactttt ttcgaggtgg ttttcagttt cggtttgaac
     4621 gtatcgtatg caatgttctg atgatgtcct ttcttgaagc tccagcgcag gaaccatttg
     4681 aggaatccca tttgcttgcc gatggtgctg tttctcatat ccttggtgtc acgcaggaag
     4741 ttgacgtatt cgttcaatcc aaactcgttg aaatagttga acgttgcatc ctccttgaac
     4801 tctttgaggt ggttcctcac tgctgcaaat ttttcatagg tggatgccgt ccagttattc
     4861 tggttaccgc actcttttac aaactcatcg aacacctccc aaaagctgac aggggcttct
     4921 tccggctgtt cttcactggt gtctttcatt ctcatgttga aagcttcctt caactgttgg
     4981 gtcgttggca tgacctcctg cacctcaaat tccttgaaaa tattctggat ttcggcatag
     5041 tatttcagca agtccgtatt gatttcggct gcactttgct ttagcttgtt ggtacatccg
     5101 ttctttaccc gctgcttatc tgcatcccat ttggctacgt caatccggta gcccgttgta
     5161 aactcgatac gttggctggc aaagatgaca cgcatacgga tgggtacgtt ctctacgatt
     5221 ggcacaccgt tctttttccg gctctccaat gcaaaaatga tgttgcgctt gatattcata
     5281 attgggtgcg tttgaaattc tacacccaaa tatacaccca attattgaga tagcaaaaga
     5341 catttagaaa catttacttt tactctatat tgtaatttac acttgattat cagtcgtttg
     5401 cagttttatg atattctgtg aaagtataag ttcgagagcc tgtctctccg caaaaaacgc
     5461 tgaaaatcag cagattgcaa aacaaacacc ctgttttaca cccaagaatg taaagtcggg
     5521 tgtttttgtt ttatttaaga taatacaacc actacataat aaaagagtag cgatattaaa
     5581 agaatccgat gagaaaagac taatatttat ctatccattc agtttgattt ctcaggactt
     5641 tacatcgtcc tgaaagtatt tgttggtacc ggtaccgagg acgcgacagc tgcagacgcg
     5701 ttatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5761 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5821 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc
     5881 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5941 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6001 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6061 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6121 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6181 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6241 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6301 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6361 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6421 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6481 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6541 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6601 atcatggccg cgggattaaa agtcggggat tggtgaacaa aaaggtgttt ctctctttaa
     6661 gagaaatatc gttttgctaa acagttgata ttgaggtatc attttatcgt aaaagacatt
     6721 tttgctcaac aattgcttga cggaaatcaa caaattttag cattttgtaa aaaagtcgct
     6781 atataatttg gtgaattgga gttattttca tatttttgca tcccgaagag tttctcttaa
     6841 agagagaaac atcttttgca taccttttcc gaccgaattt ttatgtcgta aagaggggct
     6901 ttgcaggggg tggactcaga aagatgagaa tagatgacta ttgtagttga aacacataga
     6961 aagttgctga tatacagacc gatacgcata tcgggatgaa ccatgagtac gttcttttct
     7021 caaaaaacat aaatattcga aaagagatgc aataaattaa ggagaggtta taatgaacaa
     7081 agtaaatata aaagatagtc aaaattttat tacttcaaaa tatcacatag aaaaaataat
     7141 gaattgcata agtttagatg aaaaagataa catctttgaa ataggtgcag ggaaaggtca
     7201 ttttactgct ggattggtaa agagatgtaa ttttgtaacg gcgatagaaa ttgattctaa
     7261 attatgtgag gtaactcgta ataagctctt aaattatcct aactatcaaa tagtaaatga
     7321 tgatatactg aaatttacat ttcctagcca caatccatat aaaatatttg gcaacatacc
     7381 ttacaacata agcacaaata taattcgaaa aattgttttt gaaagttcag ccacaataag
     7441 ttatttaata gtggaatatg gttttgctaa aatgttatta gatacaaaca gatcactagc
     7501 attgctgtta atggcagagg tagatatttc tatattagca aaaattccta ggtattattt
     7561 ccatccaaaa cctaaagtgg atagcacatt aattgtatta aaaagaaagc cagcaaaaat
     7621 ggcatttaaa gagagaaaaa aatatgaaac ttttgtaatg aaatgggtta acaaagagta
     7681 cgaaaaactg tttacaaaaa atcaatttaa taaagcttta aaacatgcga gaatatatga
     7741 tataaacaat attagtttcg aacaatttgt atcgctattt aatagttata aaatatttaa
     7801 cggctaaaaa caataggcca catgcaactg taaatgttta cgcgggtacc gacaccgcgg
     7861 tggaggggaa ttcccatgtc agccgttaag tgttcctgtg tcactcaaaa ttgctttgag
     7921 aggctctaag ggcttctcag tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc
     7981 ttaaaagctt taaaagcctt atatattctt ttttttctta taaaacttaa aaccttagag
     8041 gctatttaag ttgctgattt atattaattt tattgttcaa acatgagagc ttagtacgtg
     8101 aaacatgaga gcttagtacg ttagccatga gagcttagta cgttagccat gagggtttag
     8161 ttcgttaaac atgagagctt agtacgttaa acatgagagc ttagtacgtg aaacatgaga
     8221 gcttagtacg tactatcaac aggttgaact gctgatcttc agatcctcta cgccggacgc
     8281 atcgtggccg gatcaattcc gttttccgct gcataaccct gcttcggggt cattatagcg
     8341 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
     8401 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
     8461 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
     8521 aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg
     8581 gctgatgaaa ccaagccaac caggaagggc agcccaccta tcacggaatt gatccccctc
     8641 gaattggaat tctagcctgg aatagggaag caccacatca cc
//
