LOCUS       sequence-496328-        8067 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  PBT0779 biosensor circuit (pTetR85 version) in pNBU2 integrative 
            Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8067)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 8067)
  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: pEM660
FEATURES             Location/Qualifiers
     source          1..8067
                     /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    2223..2241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2262..2280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(2363..2365,2366..2368,2369..3082)
                     /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(2408..2429)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      3103..3130
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(5110..5901,5902..5970))
                     /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     5733..5752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7262..7650
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7821..7930
                     /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 aaatttaaag
     2221 tttccctatc agtgatagag aatgttctta tatttgcagt gtccctatca gtgatagaga
     2281 gatcacacaa catttaaaaa ataacattat gaaagcactc gaaaagagat tcaaaggtaa
     2341 tattaacaat aatttatttt caatggtgag caagggcgag gagctgttca ccggggtggt
     2401 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
     2461 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2521 gctgcccgtg ccctggccca ccctcgtgac caccttaggc tacggcgtgg cctgcttcgc
     2581 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2641 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt
     2701 gaagttcgag ggcgacaccc tggtgaaccg catcgtgctg aagggcatcg acttcaagga
     2761 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca aggtctatat
     2821 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagacccgcc acaacgttga
     2881 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2941 cgtgctgctg cccgacaacc actacctgag ccatcagtcc aagctgagca aagaccccaa
     3001 cgagaagcgc gatcacatgg tcctgaagga gagggtgacc gccgccggga ttacacatga
     3061 catggacgag ctgtacaagt aatagcgttc agctcgactg tgagaaggcc atcctgacgg
     3121 atggcctttt tttttcgtta taactagttg cgttcctagc cgctactagt gtaattgcct
     3181 atcttccagt gatggaacag catttgtgca ttggctgcaa caatcagcct tacttgtgcc
     3241 tgttctattt ccgaaccgac cgcttgtatg aatgcatcaa aattcgtttt ctctacgttg
     3301 gattccttgt tgctcatatt gtgatgataa tttctacaaa tatagtcatt ggtaactatc
     3361 tatgaaactg tttgatactt ttatagttga ttaaacttgt tcatggcatt tgccttaata
     3421 tcatccgcta tgtcaatgta gggtttcata gctttgtagt cgctgtgtcc cgtccatttc
     3481 atgaccacct gtgccgggat tccgagagcc agcgcattgc agatgaatgt cctttttcct
     3541 gcatgggtac tgagcaaagc gtatttgtgt gtgacttcat caatacgttc atttcccttg
     3601 tagtaggttt cccgtacagg ctcgttgatt tctgccagtt cgcccagctc tttcaggtaa
     3661 tcgttcatct tctggttgct gatgacgggc agagccatgt aattctcgaa atggatgtcc
     3721 ttgtatttgt ccagtatggc tttgctgtat ttgttcagtt caatcgtcag gctgtcggca
     3781 gtcttgactg tggttatttc gatgtggtcg gacttcacat cgcttctttt cagattgcga
     3841 acatccgaat accgcaaact cgtaaagcag cagaacagga aaacatcacg cacacgttcc
     3901 aggtattgct tatccttggg tatctggtag tctttcagct tgttcagttc atcccaagtc
     3961 aggaagatta cttttttcga ggtggttttc agtttcggtt tgaacgtatc gtatgcaatg
     4021 ttctgatgat gtcctttctt gaagctccag cgcaggaacc atttgaggaa tcccatttgc
     4081 ttgccgatgg tgctgtttct catatccttg gtgtcacgca ggaagttgac gtattcgttc
     4141 aatccaaact cgttgaaata gttgaacgtt gcatcctcct tgaactcttt gaggtggttc
     4201 ctcactgctg caaatttttc ataggtggat gccgtccagt tattctggtt accgcactct
     4261 tttacaaact catcgaacac ctcccaaaag ctgacagggg cttcttccgg ctgttcttca
     4321 ctggtgtctt tcattctcat gttgaaagct tccttcaact gttgggtcgt tggcatgacc
     4381 tcctgcacct caaattcctt gaaaatattc tggatttcgg catagtattt cagcaagtcc
     4441 gtattgattt cggctgcact ttgctttagc ttgttggtac atccgttctt tacccgctgc
     4501 ttatctgcat cccatttggc tacgtcaatc cggtagcccg ttgtaaactc gatacgttgg
     4561 ctggcaaaga tgacacgcat acggatgggt acgttctcta cgattggcac accgttcttt
     4621 ttccggctct ccaatgcaaa aatgatgttg cgcttgatat tcataattgg gtgcgtttga
     4681 aattctacac ccaaatatac acccaattat tgagatagca aaagacattt agaaacattt
     4741 acttttactc tatattgtaa tttacacttg attatcagtc gtttgcagtt ttatgatatt
     4801 ctgtgaaagt ataagttcga gagcctgtct ctccgcaaaa aacgctgaaa atcagcagat
     4861 tgcaaaacaa acaccctgtt ttacacccaa gaatgtaaag tcgggtgttt ttgttttatt
     4921 taagataata caaccactac ataataaaag agtagcgata ttaaaagaat ccgatgagaa
     4981 aagactaata tttatctatc cattcagttt gatttctcag gactttacat cgtcctgaaa
     5041 gtatttgttg gtaccggtac cgaggacgcg acagctgcag acgcgttatg agtaaacttg
     5101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5221 atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc cagatttatc
     5281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5401 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5941 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat ggccgcggga
     6001 ttaaaagtcg gggattggtg aacaaaaagg tgtttctctc tttaagagaa atatcgtttt
     6061 gctaaacagt tgatattgag gtatcatttt atcgtaaaag acatttttgc tcaacaattg
     6121 cttgacggaa atcaacaaat tttagcattt tgtaaaaaag tcgctatata atttggtgaa
     6181 ttggagttat tttcatattt ttgcatcccg aagagtttct cttaaagaga gaaacatctt
     6241 ttgcatacct tttccgaccg aatttttatg tcgtaaagag gggctttgca gggggtggac
     6301 tcagaaagat gagaatagat gactattgta gttgaaacac atagaaagtt gctgatatac
     6361 agaccgatac gcatatcggg atgaaccatg agtacgttct tttctcaaaa aacataaata
     6421 ttcgaaaaga gatgcaataa attaaggaga ggttataatg aacaaagtaa atataaaaga
     6481 tagtcaaaat tttattactt caaaatatca catagaaaaa ataatgaatt gcataagttt
     6541 agatgaaaaa gataacatct ttgaaatagg tgcagggaaa ggtcatttta ctgctggatt
     6601 ggtaaagaga tgtaattttg taacggcgat agaaattgat tctaaattat gtgaggtaac
     6661 tcgtaataag ctcttaaatt atcctaacta tcaaatagta aatgatgata tactgaaatt
     6721 tacatttcct agccacaatc catataaaat atttggcaac ataccttaca acataagcac
     6781 aaatataatt cgaaaaattg tttttgaaag ttcagccaca ataagttatt taatagtgga
     6841 atatggtttt gctaaaatgt tattagatac aaacagatca ctagcattgc tgttaatggc
     6901 agaggtagat atttctatat tagcaaaaat tcctaggtat tatttccatc caaaacctaa
     6961 agtggatagc acattaattg tattaaaaag aaagccagca aaaatggcat ttaaagagag
     7021 aaaaaaatat gaaacttttg taatgaaatg ggttaacaaa gagtacgaaa aactgtttac
     7081 aaaaaatcaa tttaataaag ctttaaaaca tgcgagaata tatgatataa acaatattag
     7141 tttcgaacaa tttgtatcgc tatttaatag ttataaaata tttaacggct aaaaacaata
     7201 ggccacatgc aactgtaaat gtttacgcgg gtaccgacac cgcggtggag gggaattccc
     7261 atgtcagccg ttaagtgttc ctgtgtcact caaaattgct ttgagaggct ctaagggctt
     7321 ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa
     7381 gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct
     7441 gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta
     7501 gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag
     7561 agcttagtac gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta
     7621 tcaacaggtt gaactgctga tcttcagatc ctctacgccg gacgcatcgt ggccggatca
     7681 attccgtttt ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat
     7741 ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg
     7801 tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt
     7861 tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga
     7921 ggctggccgg ctaccgccgg cgtaacagat gagggcaagc ggatggctga tgaaaccaag
     7981 ccaaccagga agggcagccc acctatcacg gaattgatcc ccctcgaatt ggaattctag
     8041 cctggaatag ggaagcacca catcacc
//
