LOCUS       sequence-292685-        8281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dimeric LacI repressor (LacI-11) under Rhl promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8281)
  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 8281)
  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: RA35
FEATURES             Location/Qualifiers
     source          1..8281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1137..1155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1167..1185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(1360..1389)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    1590..1608
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2248..2266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2467..2496
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2650..2668
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2680..2698
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2801..2820)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      4019..4046
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(4183..4216)
                     /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        4273..4375
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4376..5035
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4442..4461)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(5646..5673)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(5737..5770)
                     /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      5819..5853
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      6355..6577
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6625..7575
                     /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(8113..8199)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    join(8234..8281,1..184)
                     /gene="attP"
                     /label=lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
ORIGIN
        1 ttgataagca atgctttttt ataatgccaa cttagtataa aaaagctgaa cgagaaacgt
       61 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
      121 actgtaaaac acaacatatg cagtcactat gaatcaacta cttagatggt attagtgacc
      181 tgtaacagag cattagcgca aggtgatttt tgtcttcttg cgctaatttt ttgtcatcaa
      241 acctgtcgcg cactctagga gctcttatta cgctgcaagg gcgtaatttt cgtcgttcgc
      301 tgctatatat tccgggaaca ctctacaact cttttcctgc tctatatcat gaccaaagaa
      361 aataattggt ttctcttttt tcacaacttc ttttaaacgt ttaattgaag ataaagctaa
      421 ttctggatca aatcctgcga acggcacttc atcttcaaaa ttctctttcg tgtacgatgc
      481 atcaatcgtt aataaaactg aaccggattg ctccgtctca atgaatagcg actgatggcc
      541 tggagaatga cctggcgtat acaataattg aacacctggt accacttcat aatccccttc
      601 aataattttg tagttcaaat gcggtaatat acattctttc atatattctt ctctatgaag
      661 tgctgcctca tattccgttc gctgcacaat aattggtgta tttgtaaaag caccgtttcc
      721 tcctgcatga tcaaaatgta agtgagaact aataatatat aaaaggtcgt ccggctcata
      781 ccctacacgc tttaatatat tcacgattct atcttcttca gtcattttcg gtaagatctg
      841 tccttcaaca aatgtaccgt taaaaagccc ttcattatta actgcacttt ctggcatacc
      901 tgtgtctact aaaataggac cttcttccgt ctccaaaaga taacaccaca ccggcaagtt
      961 taatagtttc cccggtgtta acgcactgtt aacagacgaa tgatccaaca tgcaacgacc
     1021 tgctgggatg aaataaagtt tctttactgt catactagta tttctcctta gcatgattaa
     1081 gatgtttcag tacgaaaatt gctttcattg ttgatctcct ttttaagtgg catgcatctc
     1141 tatcactgat agggatgtgc tcagtatctc tatcactgat agggaagtaa attacagatt
     1201 tttatctttc gattacgatt tttggtttat ttcgacagtt cataggtgat tgctcaggac
     1261 atttctgtta gaaggaatcg ttttccttac ttttccttac gcacaagagt tccgtagctg
     1321 ttcaagtttg tgtttcaact gttctcgtcg tttccgcaac aagtcctctt cagaaatgag
     1381 cttttgctcc tctgcttgga cggacaggat gtatgctgtg gcttttttaa ggataactac
     1441 cttgggggcc ttttcattgt tttccaactc cgggatctgg tcacgcaggg caaaaaagct
     1501 ccgttttagc tcgttcctcc tctggcgctc caagacgttg tgtgttcgcc tcttgacatt
     1561 ctcctcggtg tccgagggcc ctgtgtgaat ctctatcact gatagggatc cacacagacg
     1621 tcttaacttg cttcctagtg aaaccgttca ggacatctac gggattgttg ctaagaaagt
     1681 caacgagatt ctacaagcag acgcaatcaa tgggaccgat aacgaagtag ttaccgtgac
     1741 cgatgagaac actggtgaaa tctctgagaa agtcaagctg ggcactaagg cactggctgg
     1801 tcaatggctg gcttacggtg ttactcgcag tgtgactaag cgttcagtca tgacgctggc
     1861 ttacgggtcc aaagagttcg gcttccgtca acaagtgctg gaagatacca ttcagccagc
     1921 tattgattcc ggcaagggtc tgatgttcac tcagccgaat caggctgctg gatacatggc
     1981 taagctgatt tgggaatctg tgagcgtgac ggtggtagct gcggttgaag caatgaactg
     2041 gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa gataagaaga ctggagagat
     2101 tcttcgcaag cgttgcgctg tgcattgggt aactcctgat ggtttccctg tgtggcagga
     2161 atacaagaag cctattcaga cgcgcttctc gagcttccca accttaccag agggcgcccc
     2221 agctggcaat tccgacgtct gtgtggatcc ctatcagtga tagagattca cacagggccc
     2281 tcggacaccg aggagaatgt caagaggcga acacacaacg tcttggagcg ccagaggagg
     2341 aacgagctaa aacggagctt ttttgccctg cgtgaccaga tcccggagtt ggaaaacaat
     2401 gaaaaggccc ccaaggtagt tatccttaaa aaagccacag catacatcct gtccgtccaa
     2461 gcagaggagc aaaagctcat ttctgaagag gacttgttgc ggaaacgacg agaacagttg
     2521 aaacacaaac ttgaacagct acggaactct tgtgcgtaag gaaaagtaag gaaaacgatt
     2581 ccttctaaca gaaatgtcct gagcaatcac ctatgaactg tcgatcctgt gaaatctggc
     2641 agttttactt ccctatcagt gatagagata ctgagcacat ccctatcagt gatagagatg
     2701 catgccactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa catcttaatc
     2761 atgctaagga gaaatactag tatgaaacca gtaacgttat acgatgtcgc agagtatgcc
     2821 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     2881 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     2941 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     3001 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     3061 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     3121 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     3181 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     3241 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     3301 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     3361 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     3421 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     3481 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     3541 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     3601 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     3661 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     3721 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     3781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgac tagtgcagcg
     3841 aacgacgaaa attacgccct tgcagcgtga ccaagcttcc cctacgtgct gctgaagttg
     3901 cccgcaacag agagtggaac caaccggtga taccacgata ctatgactga gagtcaacgc
     3961 catgagcggc ctcatttctt attctgagtt acaacagtcc gcaccgctgt ctcacactgg
     4021 ctcaccttcg ggtgggcctt tctgcgttta tatactagag agagaatata aaaagccaga
     4081 ttattaatcc ggctttttta ttatttccta gctagcagaa tccctgcttc gtccatttga
     4141 caggcacatt atgcatcgat gataagctgt caaacatgag cagaagttcc tatactttct
     4201 agagaatagg aacttcggtg tccctgttga taccgggaag ccctgggcca acttttggcg
     4261 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     4321 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     4381 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     4441 tgaggcattt cagtcagttg ctcaatgtac ctataaccag actgttcagc tggatattac
     4501 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     4561 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     4621 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     4681 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     4741 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     4801 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     4861 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     4921 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     4981 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     5041 ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa
     5101 gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt
     5161 cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt
     5221 gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta
     5281 ataattgacg atatgatcat ttattctgcc tcccagagcc tgataaaaac ggttagcgct
     5341 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     5401 ggaacataat ggtgcagggc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     5461 ggggactgtt gggcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     5521 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctaccg cctgcggccg
     5581 ctaggaaata ataaaaaagc cggattaata atctggcttt ttatattctc tctctagtat
     5641 ataaacgcag aaaggcccac ccgaaggtga gccagtgtga gacagcggtg cggactgttg
     5701 taactcagaa taagaaatga ggccgctcat ggcgttgaag ttcctatact ttctagagaa
     5761 taggaacttc ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     5821 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagggacag
     5881 taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag ccagtctgaa
     5941 tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc aggaactgct
     6001 gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc cctgagaagc
     6061 ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat aaaaggcgcc
     6121 tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac
     6181 gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt cagcgatttg
     6241 cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt ttgtagagga
     6301 gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag tagtgagtta
     6361 tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat tctataaatt
     6421 ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt
     6481 ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat acccacaact
     6541 caaaggaaaa ggactagtaa ttatcattga ctagcccatc tcaattggta tagtgattaa
     6601 aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc aaatgaacta
     6661 gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg ctgtgtggca
     6721 ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc gttcacttat
     6781 aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt attagctaaa
     6841 gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt taaaggcttt
     6901 gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga attagttttt
     6961 agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata taatctggaa
     7021 catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt attaaaagaa
     7081 ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga atttaagttc
     7141 atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa
     7201 ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga taagcgaggc
     7261 cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga tctcgtaacc
     7321 gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac cattacatca
     7381 gattcctacc tacataacgg actaagaaaa acactacacg atgctttaac tgcaaaaatt
     7441 cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagta tgatctcaat
     7501 ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat actggctaaa
     7561 tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat caagactaac
     7621 aaacaaaagt agaacaactg ttcaccgtta catatcaaag ggaaaactgt ccatatgcac
     7681 agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt ttggtgcatt
     7741 caaagctgtt caccatgaac agatcgacaa tgtaacagat gaacagcatg taacacctaa
     7801 tagaacaggt gaaaccagta aaacaaagca actagaacat gaaattgaac acctgagaca
     7861 acttgttaca gctcaacagt cacacataga cagcctgaaa caggcgatgc tgcttatcga
     7921 atcaaagctg ccgacaacac gggagccagt gacgcctccc gtggggaaaa aatcatggca
     7981 attctggaag aaatagcgct ttcagccggc aaaccggctg aagccggatc tgcgattctg
     8041 ataacaaact agcaacacca gaacagcccg tttgcgggca gcaaaacccg taccctaggt
     8101 ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     8161 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctagg gggagtggga
     8221 caaaattgaa atcaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa
     8281 a
//
