LOCUS       sequence-496395-        8066 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  PBT0779 biosensor circuit (pTetR59 version) in pNBU2 integrative 
            Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 8066)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEM658
FEATURES             Location/Qualifiers
     source          1..8066
                     /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             1474..2097
                     /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(1547..1566)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    2213..2231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(2362..2364,2365..2367,2368..3081)
                     /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(2407..2428)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      3102..3129
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(5109..5900,5901..5969))
                     /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     5732..5751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7261..7649
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7820..7929
                     /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 aaccccgagg cagcatccat tgcctgaaaa ataaccagaa
     1021 ccgagtagcg gtcttgtgca taaagcatac tttgcagttc tgacagcttt tcgatatttt
     1081 gggctaactg ttctttggca tcctgtttgg acatatcagc agtaaatgag ggattgaaat
     1141 cggatacatg atgttccttc cccggtttgg ctgtaagctc ttttaagact tcttttttca
     1201 ttgttgtata attaggttgt tataggtata taacagttga ataaagaaat cagttcttgc
     1261 caaattaaga gcgttgaata gtaattcccc aattatgtat caacagattc tacataattc
     1321 cacactaatt ccacaattat atttatattg tctgctgttt atatgtttga ttatcagtta
     1381 tatatggcgg agtgaaacgt ttggcatctg ctttgctata tattaagtac aagaaacaat
     1441 agttctatta ttcacactta aaaaattacg attatgtcca gattagataa aagtaaagtg
     1501 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
     1561 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg
     1621 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct
     1681 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct
     1741 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag
     1801 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat
     1861 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa
     1921 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta
     1981 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc
     2041 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcttaatgg
     2101 ccacatcggt tcctgctaga aggcccatcc tgacggatgg ccttttggta ttcgagcatt
     2161 acctcatact ttggattttt gttttgtttg caatggttaa tctattgtta aatccctatc
     2221 agtgatagag actttcaaat aatgttctta tatttgcagt gtcgaaagaa acaaagtagg
     2281 atcacacaac atttaaaaaa taacattatg aaagcactcg aaaagagatt caaaggtaat
     2341 attaacaata atttattttc aatggtgagc aagggcgagg agctgttcac cggggtggtg
     2401 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt ccgcggcgag
     2461 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
     2521 ctgcccgtgc cctggcccac cctcgtgacc accttaggct acggcgtggc ctgcttcgcc
     2581 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2641 gtccaggagc gcaccatctc tttcaaggac gacggtacct acaagacccg cgccgaggtg
     2701 aagttcgagg gcgacaccct ggtgaaccgc atcgtgctga agggcatcga cttcaaggag
     2761 gacggcaaca tcctggggca caagctggag tacaacttca acagccacaa ggtctatatc
     2821 acggccgaca agcagaagaa cggcatcaag gctaacttca agacccgcca caacgttgag
     2881 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2941 gtgctgctgc ccgacaacca ctacctgagc catcagtcca agctgagcaa agaccccaac
     3001 gagaagcgcg atcacatggt cctgaaggag agggtgaccg ccgccgggat tacacatgac
     3061 atggacgagc tgtacaagta atagcgttca gctcgactgt gagaaggcca tcctgacgga
     3121 tggccttttt ttttcgttat aactagttgc gttcctagcc gctactagtg taattgccta
     3181 tcttccagtg atggaacagc atttgtgcat tggctgcaac aatcagcctt acttgtgcct
     3241 gttctatttc cgaaccgacc gcttgtatga atgcatcaaa attcgttttc tctacgttgg
     3301 attccttgtt gctcatattg tgatgataat ttctacaaat atagtcattg gtaactatct
     3361 atgaaactgt ttgatacttt tatagttgat taaacttgtt catggcattt gccttaatat
     3421 catccgctat gtcaatgtag ggtttcatag ctttgtagtc gctgtgtccc gtccatttca
     3481 tgaccacctg tgccgggatt ccgagagcca gcgcattgca gatgaatgtc ctttttcctg
     3541 catgggtact gagcaaagcg tatttgtgtg tgacttcatc aatacgttca tttcccttgt
     3601 agtaggtttc ccgtacaggc tcgttgattt ctgccagttc gcccagctct ttcaggtaat
     3661 cgttcatctt ctggttgctg atgacgggca gagccatgta attctcgaaa tggatgtcct
     3721 tgtatttgtc cagtatggct ttgctgtatt tgttcagttc aatcgtcagg ctgtcggcag
     3781 tcttgactgt ggttatttcg atgtggtcgg acttcacatc gcttcttttc agattgcgaa
     3841 catccgaata ccgcaaactc gtaaagcagc agaacaggaa aacatcacgc acacgttcca
     3901 ggtattgctt atccttgggt atctggtagt ctttcagctt gttcagttca tcccaagtca
     3961 ggaagattac ttttttcgag gtggttttca gtttcggttt gaacgtatcg tatgcaatgt
     4021 tctgatgatg tcctttcttg aagctccagc gcaggaacca tttgaggaat cccatttgct
     4081 tgccgatggt gctgtttctc atatccttgg tgtcacgcag gaagttgacg tattcgttca
     4141 atccaaactc gttgaaatag ttgaacgttg catcctcctt gaactctttg aggtggttcc
     4201 tcactgctgc aaatttttca taggtggatg ccgtccagtt attctggtta ccgcactctt
     4261 ttacaaactc atcgaacacc tcccaaaagc tgacaggggc ttcttccggc tgttcttcac
     4321 tggtgtcttt cattctcatg ttgaaagctt ccttcaactg ttgggtcgtt ggcatgacct
     4381 cctgcacctc aaattccttg aaaatattct ggatttcggc atagtatttc agcaagtccg
     4441 tattgatttc ggctgcactt tgctttagct tgttggtaca tccgttcttt acccgctgct
     4501 tatctgcatc ccatttggct acgtcaatcc ggtagcccgt tgtaaactcg atacgttggc
     4561 tggcaaagat gacacgcata cggatgggta cgttctctac gattggcaca ccgttctttt
     4621 tccggctctc caatgcaaaa atgatgttgc gcttgatatt cataattggg tgcgtttgaa
     4681 attctacacc caaatataca cccaattatt gagatagcaa aagacattta gaaacattta
     4741 cttttactct atattgtaat ttacacttga ttatcagtcg tttgcagttt tatgatattc
     4801 tgtgaaagta taagttcgag agcctgtctc tccgcaaaaa acgctgaaaa tcagcagatt
     4861 gcaaaacaaa caccctgttt tacacccaag aatgtaaagt cgggtgtttt tgttttattt
     4921 aagataatac aaccactaca taataaaaga gtagcgatat taaaagaatc cgatgagaaa
     4981 agactaatat ttatctatcc attcagtttg atttctcagg actttacatc gtcctgaaag
     5041 tatttgttgg taccggtacc gaggacgcga cagctgcaga cgcgttatga gtaaacttgg
     5101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5221 tctggcccca gtgctgcaat gataccgcgg gacccacgct caccggctcc agatttatca
     5281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatcatg gccgcgggat
     6001 taaaagtcgg ggattggtga acaaaaaggt gtttctctct ttaagagaaa tatcgttttg
     6061 ctaaacagtt gatattgagg tatcatttta tcgtaaaaga catttttgct caacaattgc
     6121 ttgacggaaa tcaacaaatt ttagcatttt gtaaaaaagt cgctatataa tttggtgaat
     6181 tggagttatt ttcatatttt tgcatcccga agagtttctc ttaaagagag aaacatcttt
     6241 tgcatacctt ttccgaccga atttttatgt cgtaaagagg ggctttgcag ggggtggact
     6301 cagaaagatg agaatagatg actattgtag ttgaaacaca tagaaagttg ctgatataca
     6361 gaccgatacg catatcggga tgaaccatga gtacgttctt ttctcaaaaa acataaatat
     6421 tcgaaaagag atgcaataaa ttaaggagag gttataatga acaaagtaaa tataaaagat
     6481 agtcaaaatt ttattacttc aaaatatcac atagaaaaaa taatgaattg cataagttta
     6541 gatgaaaaag ataacatctt tgaaataggt gcagggaaag gtcattttac tgctggattg
     6601 gtaaagagat gtaattttgt aacggcgata gaaattgatt ctaaattatg tgaggtaact
     6661 cgtaataagc tcttaaatta tcctaactat caaatagtaa atgatgatat actgaaattt
     6721 acatttccta gccacaatcc atataaaata tttggcaaca taccttacaa cataagcaca
     6781 aatataattc gaaaaattgt ttttgaaagt tcagccacaa taagttattt aatagtggaa
     6841 tatggttttg ctaaaatgtt attagataca aacagatcac tagcattgct gttaatggca
     6901 gaggtagata tttctatatt agcaaaaatt cctaggtatt atttccatcc aaaacctaaa
     6961 gtggatagca cattaattgt attaaaaaga aagccagcaa aaatggcatt taaagagaga
     7021 aaaaaatatg aaacttttgt aatgaaatgg gttaacaaag agtacgaaaa actgtttaca
     7081 aaaaatcaat ttaataaagc tttaaaacat gcgagaatat atgatataaa caatattagt
     7141 ttcgaacaat ttgtatcgct atttaatagt tataaaatat ttaacggcta aaaacaatag
     7201 gccacatgca actgtaaatg tttacgcggg taccgacacc gcggtggagg ggaattccca
     7261 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
     7321 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
     7381 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
     7441 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
     7501 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
     7561 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
     7621 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcaa
     7681 ttccgttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     7741 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     7801 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     7861 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     7921 gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgat gaaaccaagc
     7981 caaccaggaa gggcagccca cctatcacgg aattgatccc cctcgaattg gaattctagc
     8041 ctggaatagg gaagcaccac atcacc
//
