LOCUS       sequence-298364-        8231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses chimeric repressor (RbsR-L) under Cin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  AUTHORS   Alnahhas RN, Sadeghpour M, Chen Y, Frey AA, Ott W, Josic K, Bennett 
            MR
  TITLE     Majority sensing in synthetic microbial consortia.
  JOURNAL   Nat Commun. 2020 Jul 21;11(1):3659. doi: 10.1038/s41467-020-17475-z.
  PUBMED    32694598
REFERENCE   2  (bases 1 to 8231)
  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"; journalName: "Nat 
            Commun."; date: "2020-07-21"; volume: "11(1):3659. doi"; pages: " 
            10.1038/s41467-020-17475-z"
COMMENT     Sequence Label: C335
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1062..1080
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1092..1110
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(1285..1314)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    1515..1533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2173..2191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2392..2421
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2575..2593
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2605..2623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2726..2745)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3894..3921
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(4058..4091)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        4148..4250
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4251..4910
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4317..4336)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(5521..5548)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(5612..5645)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     terminator      5694..5728
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      6230..6452
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6500..7450
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(7988..8074)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    join(8109..8231,1..109)
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
ORIGIN
        1 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
       61 atatgcagtc actatgaatc aactacttag atggtattag tgacctgtaa cagagcatta
      121 gcgcaaggtg atttttgtct tcttgcgcta attttttgtc atcaaacctg tcgcgcactc
      181 taggagctct tattacgctg caagggcgta attttcgtcg ttcgctgcta tatattccgg
      241 gaacactcta caactctttt cctgctctat atcatgacca aagaaaataa ttggtttctc
      301 ttttttcaca acttctttta aacgtttaat tgaagataaa gctaattctg gatcaaatcc
      361 tgcgaacggc acttcatctt caaaattctc tttcgtgtac gatgcatcaa tcgttaataa
      421 aactgaaccg gattgctccg tctcaatgaa tagcgactga tggcctggag aatgacctgg
      481 cgtatacaat aattgaacac ctggtaccac ttcataatcc ccttcaataa ttttgtagtt
      541 caaatgcggt aatatacatt ctttcatata ttcttctcta tgaagtgctg cctcatattc
      601 cgttcgctgc acaataattg gtgtatttgt aaaagcaccg tttcctcctg catgatcaaa
      661 atgtaagtga gaactaataa tatataaaag gtcgtccggc tcatacccta cacgctttaa
      721 tatattcacg attctatctt cttcagtcat tttcggtaag atctgtcctt caacaaatgt
      781 accgttaaaa agcccttcat tattaactgc actttctggc atacctgtgt ctactaaaat
      841 aggaccttct tccgtctcca aaagataaca ccacaccggc aagtttaata gtttccccgg
      901 tgttaacgca ctgttaacag acgaatgatc caacatgcaa cgacctgctg ggatgaaata
      961 aagtttcttt actgtcatac tagtatttct ccttagcatg attaagatgt ttcagtacga
     1021 aaattgcttt cattgttgat ctccttttta agtggcatgc atctctatca ctgataggga
     1081 tgtgctcagt atctctatca ctgataggga agtaaattac agatttttat ctttcgatta
     1141 cgatttttgg tttatttcga cagttcatag gtgattgctc aggacatttc tgttagaagg
     1201 aatcgttttc cttacttttc cttacgcaca agagttccgt agctgttcaa gtttgtgttt
     1261 caactgttct cgtcgtttcc gcaacaagtc ctcttcagaa atgagctttt gctcctctgc
     1321 ttggacggac aggatgtatg ctgtggcttt tttaaggata actaccttgg gggccttttc
     1381 attgttttcc aactccggga tctggtcacg cagggcaaaa aagctccgtt ttagctcgtt
     1441 cctcctctgg cgctccaaga cgttgtgtgt tcgcctcttg acattctcct cggtgtccga
     1501 gggccctgtg tgaatctcta tcactgatag ggatccacac agacgtctta acttgcttcc
     1561 tagtgaaacc gttcaggaca tctacgggat tgttgctaag aaagtcaacg agattctaca
     1621 agcagacgca atcaatggga ccgataacga agtagttacc gtgaccgatg agaacactgg
     1681 tgaaatctct gagaaagtca agctgggcac taaggcactg gctggtcaat ggctggctta
     1741 cggtgttact cgcagtgtga ctaagcgttc agtcatgacg ctggcttacg ggtccaaaga
     1801 gttcggcttc cgtcaacaag tgctggaaga taccattcag ccagctattg attccggcaa
     1861 gggtctgatg ttcactcagc cgaatcaggc tgctggatac atggctaagc tgatttggga
     1921 atctgtgagc gtgacggtgg tagctgcggt tgaagcaatg aactggctta agtctgctgc
     1981 taagctgctg gctgctgagg tcaaagataa gaagactgga gagattcttc gcaagcgttg
     2041 cgctgtgcat tgggtaactc ctgatggttt ccctgtgtgg caggaataca agaagcctat
     2101 tcagacgcgc ttctcgagct tcccaacctt accagagggc gccccagctg gcaattccga
     2161 cgtctgtgtg gatccctatc agtgatagag attcacacag ggccctcgga caccgaggag
     2221 aatgtcaaga ggcgaacaca caacgtcttg gagcgccaga ggaggaacga gctaaaacgg
     2281 agcttttttg ccctgcgtga ccagatcccg gagttggaaa acaatgaaaa ggcccccaag
     2341 gtagttatcc ttaaaaaagc cacagcatac atcctgtccg tccaagcaga ggagcaaaag
     2401 ctcatttctg aagaggactt gttgcggaaa cgacgagaac agttgaaaca caaacttgaa
     2461 cagctacgga actcttgtgc gtaaggaaaa gtaaggaaaa cgattccttc taacagaaat
     2521 gtcctgagca atcacctatg aactgtcgag ggggcctatc tgagggaatt tacttcccta
     2581 tcagtgatag agatactgag cacatcccta tcagtgatag agatgcatgc cacttaaaaa
     2641 ggagatcaac aatgaaagca attttcgtac tgaaacatct taatcatgct aaggagaaat
     2701 actagtatga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     2761 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     2821 gaagcggcga tggcggagct caattacatt cccaaccgcg tggcacaaca actggcgggc
     2881 aaagcgtcgc ataccattgg catgttgatc actgccagta ccaatccttt ctattcagaa
     2941 ctggtgcgtg gcgttgaacg cagctgcttc gaacgcggtt atagtctcgt cctttgcaat
     3001 accgaaggcg atgaacagcg gatgaatcgc aatctggaaa cgctgatgca aaaacgcgtt
     3061 gatggcttgc tgttactgtg caccgaaacg catcaacctt cgcgtgaaat catgcaacgt
     3121 tatccgacag tgcctactgt gatgatggac tgggctccgt tcgatggcga cagcgatctt
     3181 attcaggata actcgttgct gggcggagac ttagcaacgc aatatctgat cgataaaggt
     3241 catacccgta tcgcctgtat taccggcccg ctggataaaa ctccggcgcg cctgcggttg
     3301 gaaggttatc gggcggcgat gaaacgtgcg ggtctcaaca ttcctgatgg ctatgaagtc
     3361 actggtgatt ttgaatttaa cggcgggttt gacgctatgc gccaactgct atcacatccg
     3421 ctgcgtcctc aggccgtctt taccggaaat gacgctatgg ctgttggcgt ttaccaggcg
     3481 ttatatcagg cagagttaca ggttccgcag gatatcgcgg tgattggcta tgacgatatc
     3541 gaactggcaa gctttatgac gccaccatta accactatcc accaaccgaa agatgaactg
     3601 ggggagctgg cgattgatgt actcatccat cggataaccc agccgaccct tcagcaacaa
     3661 cgattacaac ttactccgat tctgatggaa cgcggttcgg ctacctccgc agcgaacgac
     3721 gaaaattacg cccttgcagc gtaataaaag cttcccctac gtgctgctga agttgcccgc
     3781 aacagagagt ggaaccaacc ggtgatacca cgatactatg actgagagtc aacgccatga
     3841 gcggcctcat ttcttattct gagttacaac agtccgcacc gctgtctcac actggctcac
     3901 cttcgggtgg gcctttctgc gtttatatac tagagagaga atataaaaag ccagattatt
     3961 aatccggctt ttttattatt tcctagctag cagaatccct gcttcgtcca tttgacaggc
     4021 acattatgca tcgatgataa gctgtcaaac atgagcagaa gttcctatac tttctagaga
     4081 ataggaactt cggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat
     4141 gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     4201 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     4261 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     4321 catttcagtc agttgctcaa tgtacctata accagactgt tcagctggat attacggcct
     4381 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     4441 cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga
     4501 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     4561 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     4621 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     4681 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     4741 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     4801 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     4861 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     4921 gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga
     4981 tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta
     5041 aatagccgct tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgac
     5101 cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat
     5161 tgacgatatg atcatttatt ctgcctccca gagcctgata aaaacggtta gcgcttcgtt
     5221 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     5281 ataatggtgc agggcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     5341 ctgttgggcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5401 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc taccgcctgc ggccgctagg
     5461 aaataataaa aaagccggat taataatctg gctttttata ttctctctct agtatataaa
     5521 cgcagaaagg cccacccgaa ggtgagccag tgtgagacag cggtgcggac tgttgtaact
     5581 cagaataaga aatgaggccg ctcatggcgt tgaagttcct atactttcta gagaatagga
     5641 acttcctcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     5701 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagg gacagtaaga
     5761 cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt ctgaatgacc
     5821 tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa ctgctgaaca
     5881 gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga gaagcccgtg
     5941 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
     6001 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
     6061 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
     6121 gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta gaggagcaaa
     6181 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
     6241 agggctggga tctattcttt ttatcttttt ttattctttc tttattctat aaattataac
     6301 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
     6361 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca caactcaaag
     6421 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
     6481 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
     6541 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
     6601 caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca cttataacca
     6661 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
     6721 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gctttgagat
     6781 tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
     6841 agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
     6901 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
     6961 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
     7021 aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
     7081 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
     7141 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
     7201 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
     7261 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
     7321 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
     7381 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
     7441 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
     7501 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
     7561 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
     7621 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
     7681 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
     7741 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
     7801 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
     7861 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
     7921 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtaccc taggtctagg
     7981 gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     8041 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ctagggggag tgggacaaaa
     8101 ttgaaatcaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     8161 taagcaatgc ttttttataa tgccaactta gtataaaaaa gctgaacgag aaacgtaaaa
     8221 tgatataaat a
//
