LOCUS       sequence-313203-        6949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert: Pfus1tet-lexA-ER-B112-tCyc1.The transcription factor 
            LexA-ER-B112 (see addgene FRP880) is inducible by alpha factor, and 
            repressible by TetR. Backbone: pRG235 (addgene). Marker: Leu2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6949)
  AUTHORS   Lormeau C, Rudolf F, Stelling J
  TITLE     A rationally engineered decoder of transient intracellular signals.
  JOURNAL   Nat Commun. 2021 Mar 25;12(1):1886. doi: 10.1038/s41467-021-22190-4.
  PUBMED    33767179
REFERENCE   2  (bases 1 to 6949)
  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: "2021-03-25"; volume: "12(1):1886. doi"; pages: " 
            10.1038/s41467-021-22190-4"
COMMENT     Sequence Label: pCL38-decoder
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(27..818,819..887))
                     /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     650..669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(888..992)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1304..2398)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFQNHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     2476..2498
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2490..2507
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2491..2507
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2514..2533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2514..2532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3296..3547,3548..3901)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     3839..3860
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             3920..4855
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             3968..4858
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      5322..5571
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5338..5356)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(5732..5750)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5771..5787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5771..5787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5784..5806)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5795..5811
                     /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)."
     primer_bind     complement(6038..6055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6209..6797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6289..6308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
       61 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
      301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
      361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
      421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat ggttgtcaga
      481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
      541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
      601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
      661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
      721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
      841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1021 gtcgaaagaa cgcggcatta gcaacatttg cgaagcgaac gattttgcga actgctatta
     1081 attaactctg gcgcgccact atcgtctatc aactaatagt tatattatca atatattatc
     1141 atatacggtg ttaagatgat gacataagtt atgagaagct gtcatcgaag ttagaggaag
     1201 ctgaagtgca aggattgata atgtaatagg ataatcatat gtttctatta tgaatttcat
     1261 ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca aggattttct
     1321 taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca cctaaatcac
     1381 cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc ttaccttctt
     1441 caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg gcgatagggt
     1501 tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac aaaccaaatg
     1561 cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc aaggaacctg
     1621 ggataacgga ggcttcatcg gagatgatgt caccaaacat gttgctggtg attataatac
     1681 catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc aattgatgtt
     1741 gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc cacagttttt ctccataatc
     1801 ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt ggctcatgtt
     1861 gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg gtgtattgtt
     1921 cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag taaatacctc
     1981 ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc ttgattggag
     2041 ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg tacaattgaa
     2101 gttctttacg gatttttagt aaaccttgtt caggtctaac actaccagta ccccatttag
     2161 gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc agcgcctcat
     2221 ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga ttttggaaat
     2281 cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg gcttcggctg
     2341 tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg gcagacatta
     2401 gaatggtata tccttgaaat atatatacga tcggatgtgc tgcaaggcga ttaagttggg
     2461 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
     2521 actcactata gggcgaattg gagaaacaga ataatatata aacctgtata atataacctt
     2581 gaagactata tttcttttcg cggccgccta tagggcgaat tggagaaaca gaataatata
     2641 taaacctgta taatataacc ttgaagacta tatttctttt cgcggccgcc tatagggcga
     2701 attggagctc cctaggacga tgattcagtt cgccttctat cctttgttta cgtatttgtt
     2761 tatatatata actttatttt tttttattaa ttgggctgca agacaatttt gttgtcagtg
     2821 atgcctcaat ccttcttttg cttccatatt taccatgtgg accctttcaa aacagagttg
     2881 tatctctgca ggatgccctt tttgacgtat tgaatggcat aattgcactg tcacttttcg
     2941 cgctgtctca ttttggtgcg atgatgaaac aaacatgaaa cgtctgtaat ttgaaacaaa
     3001 taacgtaatt ctcgggattg gttttattta aatgacaatg taagagtggc tttgtaaggt
     3061 atgtgttgct ctagaaaagg ttgaaaccag ttccctgaaa ttattcccct atctatcatt
     3121 gatagatata aaagtctatc attgatagag taattctgta aatctatttc ttaaacttct
     3181 taaattctac ttttatagtt agtctttttt ttagttttaa aacaccaaga acttagtttc
     3241 gaataaacac acataaacaa acaaaactag tggatccccc gggctgcagg aattcatgaa
     3301 agcgttaacg gccaggcaac aagaggtgtt tgatctcatc cgtgatcaca tcagccagac
     3361 aggtatgccg ccgacgcgtg cggaaatcgc gcagcgtttg gggttccgtt ccccaaacgc
     3421 ggctgaagaa catctgaagg cgctggcacg caaaggcgtt attgaaattg tttccggcgc
     3481 atcacgcggg attcgtctgt tgcaggaaga ggaagaaggg ttgccgctgg taggtcgtgt
     3541 ggctgccggt gaaccacttc tggcgcaaca gcatattgaa ggtcattatc aggtcgatcc
     3601 ttccttattc aagccgaatg ctgatttcct gctgcgcgtc agcgggatgt cgatgaaaga
     3661 catcggcatt atggatggtg acttgctggc agtgcataaa actcaggatg tacgtaacgg
     3721 tcaggtcgtt gtcgcacgta ttgatgacga ggttaccgtt aagcgcctga aaaaacaggg
     3781 caataaagtc gaactgttgc cagaaaatag cgagtttaaa ccaattgtcg tagatcttcg
     3841 tcagcagagc ttcaccattg aagggctggc ggttggggtt attcgcaacg gcgactggct
     3901 gggttctaag gatatctctg ctggagacat gagagctgcc aacctttggc caagcccgct
     3961 catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg accagatggt
     4021 cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc ctaccagacc
     4081 cttcagtgaa gcttcgatga tgggcttact gaccaacctg gcagacaggg agctggttca
     4141 catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc atgatcaggt
     4201 ccaccttcta gaatgtgcct ggttagagat cctgatgatt ggtctcgtct ggcgctccat
     4261 ggagcaccca gtgaagctac tgtttgctcc taacttgctc ttggacagga accagggaaa
     4321 atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat ctcggttccg
     4381 catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt tgcttaattc
     4441 tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg accatatcca
     4501 ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg caggcctgac
     4561 cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc acatcaggca
     4621 catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg tggtgcccct
     4681 ctatgacctg ctgctggaga tgctggacgc ccaccgccta catgcgccca ctagccgtgg
     4741 aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct ctacttcatc
     4801 gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg ccacagtcga
     4861 tatcatgcag gatctgccgg gcaacgataa cagcaccgcg ggcgaatttc caggtattac
     4921 tttgagaatc caagaaactg atatgttgta taaaggtgat actttgtatt tggattggtt
     4981 ggaagatggt attgctgaat tggttttcga tgctccaggt tctgttaata aattggatac
     5041 tgctgttgct tctttgggtg aagctattgg tgttttggaa caacaatctg atttgatttg
     5101 ggaaactttg actgttaaag atgctaaagt taatttcgat tctggtttgg aaaaattcga
     5161 agaagctatt ccatctgctg atgatttcga tccagttgct gaaagaagat cctctggtga
     5221 atttagagct gaaagacatt ctggtggtac tgatttgtgt ttcaagcttt aaacgcgttt
     5281 aaagcttatc ctcgagacag gccccttttc ctttgtctat atcatgtaat tagttatgtc
     5341 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     5401 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     5461 tttatatttc aaatttttct tttttttctg tacaaacgcg tgtacgcatg taacattata
     5521 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caaggttcgc
     5581 agtttacact ctcatcgtcg ctctcatcat cgcttccgtt gttgttttcc ttagtagcgt
     5641 ctgcttggta cccagcttcg gccgtattta cagataacga tgtcgttatt atatatatat
     5701 atatattgtc acttgtaccc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     5761 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca acagctggaa
     5821 acatataaaa cggaatgagg aataatcgta atattagtat gtagaaatat agattccatt
     5881 ttgaggattc ctatatcctt gaggagaact tctagtatat tctgtatacc taatattata
     5941 gcctttatca acaatggcgc gcctctccct gcaggcatta atgaatcggc caacgcgcgg
     6001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6781 gaaaaaaagg atctcaagaa gatcctttga tcttttgttt aaacctacgg ggtctgacgc
     6841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagt
//
