LOCUS       sequence-496326-        8698 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  PBT0779 biosensor circuit (pLacI120 version) in pNBU2 integrative 
            Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8698)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 8698)
  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: pEM663
FEATURES             Location/Qualifiers
     source          1..8698
                     /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             1483..2562
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLEET"
     primer_bind     complement(1502..1521)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      2663..2749
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2812..2828
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    2849..2865
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             join(2994..2996,2997..2999,3000..3713)
                     /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(3039..3060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      3734..3761
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(5741..6532,6533..6601))
                     /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     6364..6383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7893..8281
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            8452..8561
                     /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 attatggtga atgtgaaacc agtaacgtta
     1501 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     1561 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     1621 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     1681 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     1741 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     1801 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     1861 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     1921 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     1981 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     2041 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     2101 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     2161 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     2221 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     2281 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     2341 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     2401 ctctctcagg gccaggcggt gaagggcaat cagctgttgc cagtttcact ggtgaaaaga
     2461 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2521 atgcagctgg cacgacaggt ttcccgactg gaagagacgt aagtggagtg accttaaatc
     2581 aaggacgata ttaatcggaa ggagtattca acgtgatgaa gtcgcagggt tgacgtggga
     2641 cgtctctgat cggatctcca ggcatataaa acgaaaggct cagtcgaaag actgggcctt
     2701 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc ccaaagcact
     2761 gctcgctttg ttaagggtat tcgagcatta cctcatactt tggatttttt tttgtgagcg
     2821 gataacaagt taatctattg ttaaaatttt gtgagcggat aacaatttca aataatgttc
     2881 ttatatttgc agtgtcgaaa gaaacaaagt aggatcacac aacatttaaa aaataactta
     2941 tgaaagcact cgaaaagaga ttcaaaggta atattaacaa taatttattt tcaatggtga
     3001 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3061 taaacggcca caagttcagc gtccgcggcg agggcgaggg cgatgccacc aacggcaagc
     3121 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3181 ccaccttagg ctacggcgtg gcctgcttcg cccgctaccc cgaccacatg aagcagcacg
     3241 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc tctttcaagg
     3301 acgacggtac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3361 gcatcgtgct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3421 agtacaactt caacagccac aaggtctata tcacggccga caagcagaag aacggcatca
     3481 aggctaactt caagacccgc cacaacgttg aggacggcgg cgtgcagctc gccgaccact
     3541 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3601 gccatcagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgaagg
     3661 agagggtgac cgccgccggg attacacatg acatggacga gctgtacaag taatagcgtt
     3721 cagctcgact gtgagaaggc catcctgacg gatggccttt ttttttcgtt ataactagtt
     3781 gcgttcctag ccgctactag tgtaattgcc tatcttccag tgatggaaca gcatttgtgc
     3841 attggctgca acaatcagcc ttacttgtgc ctgttctatt tccgaaccga ccgcttgtat
     3901 gaatgcatca aaattcgttt tctctacgtt ggattccttg ttgctcatat tgtgatgata
     3961 atttctacaa atatagtcat tggtaactat ctatgaaact gtttgatact tttatagttg
     4021 attaaacttg ttcatggcat ttgccttaat atcatccgct atgtcaatgt agggtttcat
     4081 agctttgtag tcgctgtgtc ccgtccattt catgaccacc tgtgccggga ttccgagagc
     4141 cagcgcattg cagatgaatg tcctttttcc tgcatgggta ctgagcaaag cgtatttgtg
     4201 tgtgacttca tcaatacgtt catttccctt gtagtaggtt tcccgtacag gctcgttgat
     4261 ttctgccagt tcgcccagct ctttcaggta atcgttcatc ttctggttgc tgatgacggg
     4321 cagagccatg taattctcga aatggatgtc cttgtatttg tccagtatgg ctttgctgta
     4381 tttgttcagt tcaatcgtca ggctgtcggc agtcttgact gtggttattt cgatgtggtc
     4441 ggacttcaca tcgcttcttt tcagattgcg aacatccgaa taccgcaaac tcgtaaagca
     4501 gcagaacagg aaaacatcac gcacacgttc caggtattgc ttatccttgg gtatctggta
     4561 gtctttcagc ttgttcagtt catcccaagt caggaagatt acttttttcg aggtggtttt
     4621 cagtttcggt ttgaacgtat cgtatgcaat gttctgatga tgtcctttct tgaagctcca
     4681 gcgcaggaac catttgagga atcccatttg cttgccgatg gtgctgtttc tcatatcctt
     4741 ggtgtcacgc aggaagttga cgtattcgtt caatccaaac tcgttgaaat agttgaacgt
     4801 tgcatcctcc ttgaactctt tgaggtggtt cctcactgct gcaaattttt cataggtgga
     4861 tgccgtccag ttattctggt taccgcactc ttttacaaac tcatcgaaca cctcccaaaa
     4921 gctgacaggg gcttcttccg gctgttcttc actggtgtct ttcattctca tgttgaaagc
     4981 ttccttcaac tgttgggtcg ttggcatgac ctcctgcacc tcaaattcct tgaaaatatt
     5041 ctggatttcg gcatagtatt tcagcaagtc cgtattgatt tcggctgcac tttgctttag
     5101 cttgttggta catccgttct ttacccgctg cttatctgca tcccatttgg ctacgtcaat
     5161 ccggtagccc gttgtaaact cgatacgttg gctggcaaag atgacacgca tacggatggg
     5221 tacgttctct acgattggca caccgttctt tttccggctc tccaatgcaa aaatgatgtt
     5281 gcgcttgata ttcataattg ggtgcgtttg aaattctaca cccaaatata cacccaatta
     5341 ttgagatagc aaaagacatt tagaaacatt tacttttact ctatattgta atttacactt
     5401 gattatcagt cgtttgcagt tttatgatat tctgtgaaag tataagttcg agagcctgtc
     5461 tctccgcaaa aaacgctgaa aatcagcaga ttgcaaaaca aacaccctgt tttacaccca
     5521 agaatgtaaa gtcgggtgtt tttgttttat ttaagataat acaaccacta cataataaaa
     5581 gagtagcgat attaaaagaa tccgatgaga aaagactaat atttatctat ccattcagtt
     5641 tgatttctca ggactttaca tcgtcctgaa agtatttgtt ggtaccggta ccgaggacgc
     5701 gacagctgca gacgcgttat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5761 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5821 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5881 gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5941 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6001 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6061 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6121 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6181 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6241 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6301 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6361 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6421 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6481 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6541 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6601 tactcttcct ttttcaatca tggccgcggg attaaaagtc ggggattggt gaacaaaaag
     6661 gtgtttctct ctttaagaga aatatcgttt tgctaaacag ttgatattga ggtatcattt
     6721 tatcgtaaaa gacatttttg ctcaacaatt gcttgacgga aatcaacaaa ttttagcatt
     6781 ttgtaaaaaa gtcgctatat aatttggtga attggagtta ttttcatatt tttgcatccc
     6841 gaagagtttc tcttaaagag agaaacatct tttgcatacc ttttccgacc gaatttttat
     6901 gtcgtaaaga ggggctttgc agggggtgga ctcagaaaga tgagaataga tgactattgt
     6961 agttgaaaca catagaaagt tgctgatata cagaccgata cgcatatcgg gatgaaccat
     7021 gagtacgttc ttttctcaaa aaacataaat attcgaaaag agatgcaata aattaaggag
     7081 aggttataat gaacaaagta aatataaaag atagtcaaaa ttttattact tcaaaatatc
     7141 acatagaaaa aataatgaat tgcataagtt tagatgaaaa agataacatc tttgaaatag
     7201 gtgcagggaa aggtcatttt actgctggat tggtaaagag atgtaatttt gtaacggcga
     7261 tagaaattga ttctaaatta tgtgaggtaa ctcgtaataa gctcttaaat tatcctaact
     7321 atcaaatagt aaatgatgat atactgaaat ttacatttcc tagccacaat ccatataaaa
     7381 tatttggcaa cataccttac aacataagca caaatataat tcgaaaaatt gtttttgaaa
     7441 gttcagccac aataagttat ttaatagtgg aatatggttt tgctaaaatg ttattagata
     7501 caaacagatc actagcattg ctgttaatgg cagaggtaga tatttctata ttagcaaaaa
     7561 ttcctaggta ttatttccat ccaaaaccta aagtggatag cacattaatt gtattaaaaa
     7621 gaaagccagc aaaaatggca tttaaagaga gaaaaaaata tgaaactttt gtaatgaaat
     7681 gggttaacaa agagtacgaa aaactgttta caaaaaatca atttaataaa gctttaaaac
     7741 atgcgagaat atatgatata aacaatatta gtttcgaaca atttgtatcg ctatttaata
     7801 gttataaaat atttaacggc taaaaacaat aggccacatg caactgtaaa tgtttacgcg
     7861 ggtaccgaca ccgcggtgga ggggaattcc catgtcagcc gttaagtgtt cctgtgtcac
     7921 tcaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct ggcttgttgt
     7981 ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt ttcttataaa
     8041 acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt gttcaaacat
     8101 gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc ttagtacgtt
     8161 agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat gagagcttag
     8221 tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgctg atcttcagat
     8281 cctctacgcc ggacgcatcg tggccggatc aattccgttt tccgctgcat aaccctgctt
     8341 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     8401 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     8461 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     8521 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg gcgtaacaga
     8581 tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc cacctatcac
     8641 ggaattgatc cccctcgaat tggaattcta gcctggaata gggaagcacc acatcacc
//
