LOCUS       sequence-463646-        6493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Lux activation by 3OC6-HSL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6493)
  AUTHORS   Kokarakis EJ, Rillema R, Ducat DC, Sakkos JK
  TITLE     Developing Cyanobacterial Quorum Sensing Toolkits: Toward 
            Interspecies Coordination in Mixed Autotroph/Heterotroph 
            Communities.
  JOURNAL   ACS Synth Biol. 2023 Jan 20;12(1):265-276. doi: 
            10.1021/acssynbio.2c00527. Epub 2022 Dec 27.
  PUBMED    36573789
REFERENCE   2  (bases 1 to 6493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.2c00527"; journalName: "ACS Synth Biol"; date: 
            "2023-01-20- 20"; volume: "12"; issue: "1"; pages: "265-276"
COMMENT     Sequence Label: pNS3_Ptrc::luxR _Plux::mNG
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      82..168
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      260..287
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(320..325,326..343,344..350)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    358..374
                     /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)."
     RBS             1313..1324
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        2065..2166
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2167..2826
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2233..2252)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        2891..2968
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2969..4051
                     /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
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2988..3007)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      complement(5033..5621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5113..5132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccgtggctc ccagcgatcg agcgcttcag ctgtggccgc caccaggtca acgatgaggg
       61 actggttgat cggctagtca tcaaataaaa cgaaaggctc agtcggaaga ctgggccttt
      121 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
      181 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
      241 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctactc
      301 tagtttcgaa accccaggct tgacacttta tgcttccggc tcgtataatg tgtggaattg
      361 tgagcggata acaatttcac acaaggagga aaaacatatg tctagaatga agaatattaa
      421 cgcagacgac acatatcgaa tcattaacaa gattaaggcg tgtcgcagta acaatgacat
      481 caatcagtgc ttgagtgata tgacgaaaat ggttcattgt gagtattact tactagccat
      541 tatttatccg cactccatgg tcaagagcga cattagcatt ctggacaatt acccaaagaa
      601 atggcggcaa tactacgacg acgcaaacct cattaagtat gacccaatcg tagattacag
      661 taattccaac cacagtccga ttaactggaa tatcttcgaa aataatgccg tgaacaaaaa
      721 atccccgaac gtaatcaagg aagctaagac aagtggtctc attacgggat ttagtttccc
      781 aatccacaca gcaaacaatg ggtttggaat gctgtctttt gctcatagtg agaaagataa
      841 ctatatcgac tccctgtttt tgcatgcttg tatgaacatt ccactcattg tgccatcttt
      901 agtagataac tatcgcaaga ttaatatcgc caataataaa agcaataatg atctcactaa
      961 acgggaaaag gagtgtctag cttgggcgtg tgaaggaaaa tcttcttggg acatttctaa
     1021 gattctcggt tgttctgaac gcacggttac gtttcattta acgaatgcgc agatgaaact
     1081 gaatactaca aaccgttgtc agagtatctc caaagcgatc ctgaccgggg ccattgactg
     1141 tccttacttt aagaattgat aactcggtac caaattccag aaaagagacg ctgaaaagcg
     1201 tcttttttcg ttttggtcca ctattgtatc gctgggaata caattactta acataagcac
     1261 ctgtaggatc gtacaggttt acgcaagaaa atggtttgtt atagtcgaat ataaagagga
     1321 gaaaggtacc atggtcagca aaggtgaaga agacaatatg gcgagcctgc cggcgacgca
     1381 tgaactgcac attttcggta gcatcaacgg tgtggacttc gatatggttg gccagggtac
     1441 cggcaacccg aatgacggtt atgaagaact gaatctgaaa agtacgaagg gcgatctgca
     1501 attttccccg tggattctgg tcccgcatat cggttacggc ttccaccagt acctgccgta
     1561 tccggatggt atgagcccgt ttcaagcggc catggtggac ggttctggct atcaggttca
     1621 tcgtaccatg caattcgaag atggcgcgag tctgaccgtg aactatcgct acacgtatga
     1681 aggttcacac attaaaggcg aagcgcaggt caaaggtacc ggttttccgg cggatggtcc
     1741 ggtgatgacc aactcactga cggcagctga ttggtgccgt tcgaaaaaga cctatccgaa
     1801 tgacaaaacc attatcagca cgtttaagtg gtcttacacc acgggtaacg gcaaacgtta
     1861 tcgcagcacc gcacgtacca cgtacacgtt cgctaaaccg atggcggcca actacctgaa
     1921 aaatcagccg atgtatgttt ttcgcaaaac cgaactgaaa cattccaaga cggaactgaa
     1981 cttcaaggaa tggcaaaagg cgtttacgga tgtgatgggt atggacgaac tgtacaagta
     2041 aaagcttgcg gccgcgtagg atccgatcgg cacgtaagag gttccaactt tcaccataat
     2101 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     2161 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     2221 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     2281 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     2341 tttattcaca ttcttgcccg cctgatgaat gctcatccgg agttccgtat ggcaatgaaa
     2401 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     2461 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     2521 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     2581 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     2641 aacgtggcca atatggacaa cttcttcgcc cccgttttca ctatgggcaa atattatacg
     2701 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     2761 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     2821 gcgtaatttt tttaaggcag ttattggtgc ccttgaattc ctactagtcg aagcggcatg
     2881 catttacgtt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
     2941 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     3001 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     3061 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     3121 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     3181 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     3241 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     3301 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     3361 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     3421 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     3481 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     3541 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     3601 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     3661 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     3721 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     3781 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     3841 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     3901 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     3961 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4021 acaggtttcc cgactggaaa gcgggcagtg atcccacagc cgccagttcc gctggcggca
     4081 ttttaacttt ctttaataat ctagtgacaa gccggggcag acgtgagccg tagtcccgtc
     4141 gccagacgcg ggtgcccacg ggcgtcgtca ggatgtccgt aattgactgc cggaggtggt
     4201 caatgccctt cagctccttg ccactgtcac ggctcatgcc tcgggtcatt agtcgcccgc
     4261 tccggtatct tcactggctt cgatgattgc cgccccgcag ctgcagaggt caccgatccg
     4321 agcagtcggc ctctggttgg taaagaccgt gcgactgccg gtgatgatcg tgttcaagcc
     4381 atgcaggggg caggcgtgaa ggtcatcctt tcgggccacg gggcggctgt tgacgaaggt
     4441 gtcgtcgctc ccggtgatga tgatcccgcc gtgatcggtc acgtcgttta gtcgagcgat
     4501 gcctggcgtc gtagtcacgg gtttaggtca atacgacttg cggtcactgt aacgttgccc
     4561 tcggcggtca cgttaacgtc gccttgggct tcgacttgcg cctcctgcac aaggatcaca
     4621 atccgtcctt gggctgcggt gaggtcgatc ttgtactcat gcgcttcgcg gtcgtactgg
     4681 atgattgagt catcctcgaa ctgcgtcttt tggatcgttt ctttgtcctc gatctggggg
     4741 tagtcagtcg agaacgcgcc gggcatcgcg aagccctgac tgatctcgcc ggagggggcc
     4801 atcacgacga cggcctcacc gacctcgggc gcccaccaga accgatcctt gcccgctcgc
     4861 tgcgtgagcc acggaatcca gtcagtgagg agcagcgcct cgccgctctc ctcgtcttcc
     4921 tcgatcgcga cacggatcag ccccttggga tagtcagcct cggctaccct gcctacgcgg
     4981 agcaagttgc cgtgacgccg actgtctcga gtctagtcgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5641 ctacggggtc tgacgctcag tctagttcag ccagctcgtc gtgatgtcga aacccaagcc
     5701 accctcagag gtgaaggccg cttcgagcac atcgggaagc gtgtcgacga cggtgctcgg
     5761 tgcctcgggt aagagccaga tagatgcggt gacgttcacc gtcgcgatcg tggcgggaac
     5821 cacggtcact gcgtccgtta caactcgcac gtcgtcggcc agcacgactg actcgacggc
     5881 ttcgattagc ccgggagagg caaggccatc aggctcggta gacagaatgc tgattaacac
     5941 ctcgccggga gcagggctgc tcacagccgc gtccttcacc cgtgggtcag ccgtcagcgc
     6001 ttgatagcgg taccaggccg caccgcctgc ggtcgagctg cccttgatcc gctcgatggt
     6061 gcgatcgcga agctcgccat ccggctcatt cggctgccgc accacgccgt agaacgcggc
     6121 aaggttgtcg aggtcggggc cgtttgagta gcgaagcagt gtcgcaagaa gtgcgtcgtt
     6181 gatccgctga cgcaggatca gctcgcgggc tgcgcagacc tccagcagct tgatgaccgg
     6241 gtcggattcg aggatggcgg tgtagctggc gtcacgcgat cgcaggtcgt cgatcaagtc
     6301 ctgcaggatc agttcaaagt ccagcgcttc gatgatggtg ggcgcgggaa tcgtagcaaa
     6361 gtcaagaacg gtcatgagac gactaagccc tccagcgtga tacgcctgcc ctcggggatg
     6421 tagtagccga tcaggttcag ctcgacttga ccagctgcgc tggctgagac gatgcgaacc
     6481 ttctccagct tca
//
