LOCUS       sequence-496325-        8699 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  PBT0779 biosensor circuit (pLacI69 version) in pNBU2 integrative 
            Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8699)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 8699)
  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: pEM665
FEATURES             Location/Qualifiers
     source          1..8699
                     /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    2850..2866
                     /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    2887..2903
                     /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(2995..2997,2998..3000,3001..3714)
                     /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(3040..3061)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      3735..3762
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(5742..6533,6534..6602))
                     /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     6365..6384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7894..8282
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            8453..8562
                     /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 tttgttttgt
     2821 ttgcaatggt taatctattg ttaaaatttt tgtgagcgga taacaattca aataatgttc
     2881 ttatatttgt gagcggataa caaacaaagt aggatcacac aacatttaaa aaataacatt
     2941 atgaaagcac tcgaaaagag attcaaaggt aatattaaca ataatttatt ttcaatggtg
     3001 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3061 gtaaacggcc acaagttcag cgtccgcggc gagggcgagg gcgatgccac caacggcaag
     3121 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3181 accaccttag gctacggcgt ggcctgcttc gcccgctacc ccgaccacat gaagcagcac
     3241 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctctttcaag
     3301 gacgacggta cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3361 cgcatcgtgc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3421 gagtacaact tcaacagcca caaggtctat atcacggccg acaagcagaa gaacggcatc
     3481 aaggctaact tcaagacccg ccacaacgtt gaggacggcg gcgtgcagct cgccgaccac
     3541 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3601 agccatcagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgaag
     3661 gagagggtga ccgccgccgg gattacacat gacatggacg agctgtacaa gtaatagcgt
     3721 tcagctcgac tgtgagaagg ccatcctgac ggatggcctt tttttttcgt tataactagt
     3781 tgcgttccta gccgctacta gtgtaattgc ctatcttcca gtgatggaac agcatttgtg
     3841 cattggctgc aacaatcagc cttacttgtg cctgttctat ttccgaaccg accgcttgta
     3901 tgaatgcatc aaaattcgtt ttctctacgt tggattcctt gttgctcata ttgtgatgat
     3961 aatttctaca aatatagtca ttggtaacta tctatgaaac tgtttgatac ttttatagtt
     4021 gattaaactt gttcatggca tttgccttaa tatcatccgc tatgtcaatg tagggtttca
     4081 tagctttgta gtcgctgtgt cccgtccatt tcatgaccac ctgtgccggg attccgagag
     4141 ccagcgcatt gcagatgaat gtcctttttc ctgcatgggt actgagcaaa gcgtatttgt
     4201 gtgtgacttc atcaatacgt tcatttccct tgtagtaggt ttcccgtaca ggctcgttga
     4261 tttctgccag ttcgcccagc tctttcaggt aatcgttcat cttctggttg ctgatgacgg
     4321 gcagagccat gtaattctcg aaatggatgt ccttgtattt gtccagtatg gctttgctgt
     4381 atttgttcag ttcaatcgtc aggctgtcgg cagtcttgac tgtggttatt tcgatgtggt
     4441 cggacttcac atcgcttctt ttcagattgc gaacatccga ataccgcaaa ctcgtaaagc
     4501 agcagaacag gaaaacatca cgcacacgtt ccaggtattg cttatccttg ggtatctggt
     4561 agtctttcag cttgttcagt tcatcccaag tcaggaagat tacttttttc gaggtggttt
     4621 tcagtttcgg tttgaacgta tcgtatgcaa tgttctgatg atgtcctttc ttgaagctcc
     4681 agcgcaggaa ccatttgagg aatcccattt gcttgccgat ggtgctgttt ctcatatcct
     4741 tggtgtcacg caggaagttg acgtattcgt tcaatccaaa ctcgttgaaa tagttgaacg
     4801 ttgcatcctc cttgaactct ttgaggtggt tcctcactgc tgcaaatttt tcataggtgg
     4861 atgccgtcca gttattctgg ttaccgcact cttttacaaa ctcatcgaac acctcccaaa
     4921 agctgacagg ggcttcttcc ggctgttctt cactggtgtc tttcattctc atgttgaaag
     4981 cttccttcaa ctgttgggtc gttggcatga cctcctgcac ctcaaattcc ttgaaaatat
     5041 tctggatttc ggcatagtat ttcagcaagt ccgtattgat ttcggctgca ctttgcttta
     5101 gcttgttggt acatccgttc tttacccgct gcttatctgc atcccatttg gctacgtcaa
     5161 tccggtagcc cgttgtaaac tcgatacgtt ggctggcaaa gatgacacgc atacggatgg
     5221 gtacgttctc tacgattggc acaccgttct ttttccggct ctccaatgca aaaatgatgt
     5281 tgcgcttgat attcataatt gggtgcgttt gaaattctac acccaaatat acacccaatt
     5341 attgagatag caaaagacat ttagaaacat ttacttttac tctatattgt aatttacact
     5401 tgattatcag tcgtttgcag ttttatgata ttctgtgaaa gtataagttc gagagcctgt
     5461 ctctccgcaa aaaacgctga aaatcagcag attgcaaaac aaacaccctg ttttacaccc
     5521 aagaatgtaa agtcgggtgt ttttgtttta tttaagataa tacaaccact acataataaa
     5581 agagtagcga tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt
     5641 ttgatttctc aggactttac atcgtcctga aagtatttgt tggtaccggt accgaggacg
     5701 cgacagctgc agacgcgtta tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5881 cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6601 atactcttcc tttttcaatc atggccgcgg gattaaaagt cggggattgg tgaacaaaaa
     6661 ggtgtttctc tctttaagag aaatatcgtt ttgctaaaca gttgatattg aggtatcatt
     6721 ttatcgtaaa agacattttt gctcaacaat tgcttgacgg aaatcaacaa attttagcat
     6781 tttgtaaaaa agtcgctata taatttggtg aattggagtt attttcatat ttttgcatcc
     6841 cgaagagttt ctcttaaaga gagaaacatc ttttgcatac cttttccgac cgaattttta
     6901 tgtcgtaaag aggggctttg cagggggtgg actcagaaag atgagaatag atgactattg
     6961 tagttgaaac acatagaaag ttgctgatat acagaccgat acgcatatcg ggatgaacca
     7021 tgagtacgtt cttttctcaa aaaacataaa tattcgaaaa gagatgcaat aaattaagga
     7081 gaggttataa tgaacaaagt aaatataaaa gatagtcaaa attttattac ttcaaaatat
     7141 cacatagaaa aaataatgaa ttgcataagt ttagatgaaa aagataacat ctttgaaata
     7201 ggtgcaggga aaggtcattt tactgctgga ttggtaaaga gatgtaattt tgtaacggcg
     7261 atagaaattg attctaaatt atgtgaggta actcgtaata agctcttaaa ttatcctaac
     7321 tatcaaatag taaatgatga tatactgaaa tttacatttc ctagccacaa tccatataaa
     7381 atatttggca acatacctta caacataagc acaaatataa ttcgaaaaat tgtttttgaa
     7441 agttcagcca caataagtta tttaatagtg gaatatggtt ttgctaaaat gttattagat
     7501 acaaacagat cactagcatt gctgttaatg gcagaggtag atatttctat attagcaaaa
     7561 attcctaggt attatttcca tccaaaacct aaagtggata gcacattaat tgtattaaaa
     7621 agaaagccag caaaaatggc atttaaagag agaaaaaaat atgaaacttt tgtaatgaaa
     7681 tgggttaaca aagagtacga aaaactgttt acaaaaaatc aatttaataa agctttaaaa
     7741 catgcgagaa tatatgatat aaacaatatt agtttcgaac aatttgtatc gctatttaat
     7801 agttataaaa tatttaacgg ctaaaaacaa taggccacat gcaactgtaa atgtttacgc
     7861 gggtaccgac accgcggtgg aggggaattc ccatgtcagc cgttaagtgt tcctgtgtca
     7921 ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg
     7981 tccacaaccg ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa
     8041 aacttaaaac cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca
     8101 tgagagctta gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt
     8161 tagccatgag ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta
     8221 gtacgtgaaa catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga
     8281 tcctctacgc cggacgcatc gtggccggat caattccgtt ttccgctgca taaccctgct
     8341 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     8401 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     8461 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     8521 tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag
     8581 atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca
     8641 cggaattgat ccccctcgaa ttggaattct agcctggaat agggaagcac cacatcacc
//
