LOCUS       sequence-377793-        6450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Las activation by 3OC12-HSL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  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 6450)
  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::lasR_Plas::mNG
FEATURES             Location/Qualifiers
     source          1..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1087..1675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1167..1186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2629..2715
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2807..2834
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(2867..2872,2873..2890,2891..2897)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2905..2921
                     /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             3817..3828
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        4569..4670
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4671..5330
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4737..4756)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        5395..5472
                     /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             join(5473..6450,1..105)
                     /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(5492..5511)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
       61 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgatccca cagccgccag
      121 ttccgctggc ggcattttaa ctttctttaa taatctagtg acaagccggg gcagacgtga
      181 gccgtagtcc cgtcgccaga cgcgggtgcc cacgggcgtc gtcaggatgt ccgtaattga
      241 ctgccggagg tggtcaatgc ccttcagctc cttgccactg tcacggctca tgcctcgggt
      301 cattagtcgc ccgctccggt atcttcactg gcttcgatga ttgccgcccc gcagctgcag
      361 aggtcaccga tccgagcagt cggcctctgg ttggtaaaga ccgtgcgact gccggtgatg
      421 atcgtgttca agccatgcag ggggcaggcg tgaaggtcat cctttcgggc cacggggcgg
      481 ctgttgacga aggtgtcgtc gctcccggtg atgatgatcc cgccgtgatc ggtcacgtcg
      541 tttagtcgag cgatgcctgg cgtcgtagtc acgggtttag gtcaatacga cttgcggtca
      601 ctgtaacgtt gccctcggcg gtcacgttaa cgtcgccttg ggcttcgact tgcgcctcct
      661 gcacaaggat cacaatccgt ccttgggctg cggtgaggtc gatcttgtac tcatgcgctt
      721 cgcggtcgta ctggatgatt gagtcatcct cgaactgcgt cttttggatc gtttctttgt
      781 cctcgatctg ggggtagtca gtcgagaacg cgccgggcat cgcgaagccc tgactgatct
      841 cgccggaggg ggccatcacg acgacggcct caccgacctc gggcgcccac cagaaccgat
      901 ccttgcccgc tcgctgcgtg agccacggaa tccagtcagt gaggagcagc gcctcgccgc
      961 tctcctcgtc ttcctcgatc gcgacacgga tcagcccctt gggatagtca gcctcggcta
     1021 ccctgcctac gcggagcaag ttgccgtgac gccgactgtc tcgagtctag tcgcgttgct
     1081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1501 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1681 tcctttgatc ttttctacgg ggtctgacgc tcagtctagt tcagccagct cgtcgtgatg
     1741 tcgaaaccca agccaccctc agaggtgaag gccgcttcga gcacatcggg aagcgtgtcg
     1801 acgacggtgc tcggtgcctc gggtaagagc cagatagatg cggtgacgtt caccgtcgcg
     1861 atcgtggcgg gaaccacggt cactgcgtcc gttacaactc gcacgtcgtc ggccagcacg
     1921 actgactcga cggcttcgat tagcccggga gaggcaaggc catcaggctc ggtagacaga
     1981 atgctgatta acacctcgcc gggagcaggg ctgctcacag ccgcgtcctt cacccgtggg
     2041 tcagccgtca gcgcttgata gcggtaccag gccgcaccgc ctgcggtcga gctgcccttg
     2101 atccgctcga tggtgcgatc gcgaagctcg ccatccggct cattcggctg ccgcaccacg
     2161 ccgtagaacg cggcaaggtt gtcgaggtcg gggccgtttg agtagcgaag cagtgtcgca
     2221 agaagtgcgt cgttgatccg ctgacgcagg atcagctcgc gggctgcgca gacctccagc
     2281 agcttgatga ccgggtcgga ttcgaggatg gcggtgtagc tggcgtcacg cgatcgcagg
     2341 tcgtcgatca agtcctgcag gatcagttca aagtccagcg cttcgatgat ggtgggcgcg
     2401 ggaatcgtag caaagtcaag aacggtcatg agacgactaa gccctccagc gtgatacgcc
     2461 tgccctcggg gatgtagtag ccgatcaggt tcagctcgac ttgaccagct gcgctggctg
     2521 agacgatgcg aaccttctcc agcttcagcc gtggctccca gcgatcgagc gcttcagctg
     2581 tggccgccac caggtcaacg atgagggact ggttgatcgg ctagtcatca aataaaacga
     2641 aaggctcagt cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2701 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2761 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     2821 acggatggcc tttttgcgtt tctactctag tttcgaaacc ccaggcttga cactttatgc
     2881 ttccggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca aggaggaaaa
     2941 acatatgtct agaatggctt tagtagatgg tttcttggaa ctcgaacgta gtagtggaaa
     3001 gctagaatgg tccgctattt tacaaaagat ggcgtccgac ttgggctttt ccaaaatcct
     3061 attcgggtta ttacctaagg attcccagga ttacgaaaat gcattcattg taggcaacta
     3121 tcccgccgct tggcgagagc attatgatcg agcgggctat gcccgagtgg acccgacggt
     3181 ctctcattgc acccaaagtg tattgcctat cttttgggaa ccaagcatct atcagactcg
     3241 aaaacagcac gagttcttcg aggaagccag cgccgcaggg ttagtctatg gtttgacaat
     3301 gccgttacat ggggcgcgtg gagagttagg tgcattaagc ctatccgtag aagcagaaaa
     3361 ccgcgctgag gcgaaccgct ttattgaaag cgtgttaccc acactatgga tgttgaaaga
     3421 ttacgccttg cagtctggag ctgggttggc gttcgagcac cctgtgagca aaccagtagt
     3481 gctgacaagc cgtgaaaagg aagtattgca gtggtgtgca attggaaaaa cctcctggga
     3541 gattagcgta atttgcaact gtagcgaggc taatgttaat ttccacatgg gtaacatccg
     3601 gcgtaagttc ggcgtcacta gccgtcgtgt cgctgccatc atggccgtaa acctggggct
     3661 gattacgttg tgataactcg gtaccaaatt ccagaaaaga gacgctgaaa agcgtctttt
     3721 ttcgttttgg tccttcgagc ctagcaaggg tccgggttca ccgaaatcta tctcatttgc
     3781 tagttataaa attatgaaat ttgcgtaaat tcttcaaaag aggagaaagg taccatggtc
     3841 agcaaaggtg aagaagacaa tatggcgagc ctgccggcga cgcatgaact gcacattttc
     3901 ggtagcatca acggtgtgga cttcgatatg gttggccagg gtaccggcaa cccgaatgac
     3961 ggttatgaag aactgaatct gaaaagtacg aagggcgatc tgcaattttc cccgtggatt
     4021 ctggtcccgc atatcggtta cggcttccac cagtacctgc cgtatccgga tggtatgagc
     4081 ccgtttcaag cggccatggt ggacggttct ggctatcagg ttcatcgtac catgcaattc
     4141 gaagatggcg cgagtctgac cgtgaactat cgctacacgt atgaaggttc acacattaaa
     4201 ggcgaagcgc aggtcaaagg taccggtttt ccggcggatg gtccggtgat gaccaactca
     4261 ctgacggcag ctgattggtg ccgttcgaaa aagacctatc cgaatgacaa aaccattatc
     4321 agcacgttta agtggtctta caccacgggt aacggcaaac gttatcgcag caccgcacgt
     4381 accacgtaca cgttcgctaa accgatggcg gccaactacc tgaaaaatca gccgatgtat
     4441 gtttttcgca aaaccgaact gaaacattcc aagacggaac tgaacttcaa ggaatggcaa
     4501 aaggcgttta cggatgtgat gggtatggac gaactgtaca agtaaaagct tgcggccgcg
     4561 taggatccga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     4621 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     4681 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     4741 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     4801 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     4861 cccgcctgat gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga
     4921 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     4981 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     5041 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     5101 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     5161 acaacttctt cgcccccgtt ttcactatgg gcaaatatta tacgcaaggc gacaaggtgc
     5221 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     5281 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     5341 gcagttattg gtgcccttga attcctacta gtcgaagcgg catgcattta cgttgacacc
     5401 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     5461 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     5521 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     5581 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     5641 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     5701 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     5761 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     5821 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     5881 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     5941 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     6001 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     6061 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     6121 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     6181 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     6241 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     6301 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     6361 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     6421 cagctgttgc ccgtctcact ggtgaaaaga
//
