LOCUS       sequence-313198-        7109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert: PlexAtet-lexA-ER-B112-tCyc1. The transcription factor 
            LexA-ER-B112 (see addgene FRP880) is self-inducible upon addition of
            estradiol, and repressible by TetR. Backbone: pRG206MX. Marker: 
            Ura3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7109)
  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 7109)
  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: pCL28-decoder
FEATURES             Location/Qualifiers
     source          1..7109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(476..727,728..1081)
                     /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     1019..1040
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             1100..2035
                     /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             1148..2038
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      2502..2751
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2518..2536)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(2914..2932)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2953..2969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2953..2969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2966..2988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2977..2993
                     /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(3303..3320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3474..4062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3554..3573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4241..5032,5033..5101))
                     /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     4864..4883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5102..5206)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(5571..5768)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(6587..6930)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     7018..7040
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7032..7049
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7033..7049
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7056..7075
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7056..7074
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ataatataac cttgaagact atatttcttt tcgcggccgc ctatagggcg aattggagct
       61 ccctaggtgc tgtatatact cacagcataa ctgtatatac acccagggtc taggtgctgt
      121 atatactcac agcataactg tatatacacc cagggtctag gtgctgtata tactcacagc
      181 ataactgtat atacacccag ggtctaggtg ctgtatatac tcacagcata actgtatata
      241 cacccagggt ctagaaaagg ttgaaaccag ttccctgaaa ttattcccct atctatcatt
      301 gatagatata aaagtctatc attgatagag taattctgta aatctatttc ttaaacttct
      361 taaattctac ttttatagtt agtctttttt ttagttttaa aacaccaaga acttagtttc
      421 gaataaacac acataaacaa acaaaactag tggatccccc gggctgcagg aattcatgaa
      481 agcgttaacg gccaggcaac aagaggtgtt tgatctcatc cgtgatcaca tcagccagac
      541 aggtatgccg ccgacgcgtg cggaaatcgc gcagcgtttg gggttccgtt ccccaaacgc
      601 ggctgaagaa catctgaagg cgctggcacg caaaggcgtt attgaaattg tttccggcgc
      661 atcacgcggg attcgtctgt tgcaggaaga ggaagaaggg ttgccgctgg taggtcgtgt
      721 ggctgccggt gaaccacttc tggcgcaaca gcatattgaa ggtcattatc aggtcgatcc
      781 ttccttattc aagccgaatg ctgatttcct gctgcgcgtc agcgggatgt cgatgaaaga
      841 catcggcatt atggatggtg acttgctggc agtgcataaa actcaggatg tacgtaacgg
      901 tcaggtcgtt gtcgcacgta ttgatgacga ggttaccgtt aagcgcctga aaaaacaggg
      961 caataaagtc gaactgttgc cagaaaatag cgagtttaaa ccaattgtcg tagatcttcg
     1021 tcagcagagc ttcaccattg aagggctggc ggttggggtt attcgcaacg gcgactggct
     1081 gggttctaag gatatctctg ctggagacat gagagctgcc aacctttggc caagcccgct
     1141 catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg accagatggt
     1201 cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc ctaccagacc
     1261 cttcagtgaa gcttcgatga tgggcttact gaccaacctg gcagacaggg agctggttca
     1321 catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc atgatcaggt
     1381 ccaccttcta gaatgtgcct ggttagagat cctgatgatt ggtctcgtct ggcgctccat
     1441 ggagcaccca gtgaagctac tgtttgctcc taacttgctc ttggacagga accagggaaa
     1501 atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat ctcggttccg
     1561 catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt tgcttaattc
     1621 tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg accatatcca
     1681 ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg caggcctgac
     1741 cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc acatcaggca
     1801 catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg tggtgcccct
     1861 ctatgacctg ctgctggaga tgctggacgc ccaccgccta catgcgccca ctagccgtgg
     1921 aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct ctacttcatc
     1981 gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg ccacagtcga
     2041 tatcatgcag gatctgccgg gcaacgataa cagcaccgcg ggcgaatttc caggtattac
     2101 tttgagaatc caagaaactg atatgttgta taaaggtgat actttgtatt tggattggtt
     2161 ggaagatggt attgctgaat tggttttcga tgctccaggt tctgttaata aattggatac
     2221 tgcggttgct tctttgggtg aagctattgg tgttttggaa caacaatctg atttgatttg
     2281 ggaaactttg actgttaaag atgctaaagt taatttcgat tctggtttgg aaaaattcga
     2341 agaagctatt ccatctgctg atgatttcga tccagttgct gaaagaagat cctctggtga
     2401 atttagagct gaaagacatt ctggtggtac tgatttgtgt ttcaagcttt aaacgcgttt
     2461 aaagcttatc ctcgagacag gccccttttc ctttgtctat atcatgtaat tagttatgtc
     2521 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     2581 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     2641 tttatatttc aaatttttct tttttttctg tacaaacgcg tgtacgcatg taacattata
     2701 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caaggttcgc
     2761 agtttacact ctcatcgtcg ctctcatcat cgcttccgtt gttgttttcc ttagtagcgt
     2821 ctgcttggta cccagcttcg gccgtattta cagataacga tgtcgttatt atatatatat
     2881 atatatattg tcacttgtac ccagcttttg ttccctttag tgagggttaa tttcgagctt
     2941 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caacagctgg
     3001 atgaattgaa tagctttatg gaccctgaaa ccacagccac attaaccttc tttgatggtc
     3061 aaaacttatc cttcaccata aatatgcctc gcaaaaaagg taattaacat atatagaatt
     3121 acattattta tgaaatatca tcactatctc ttagcatctt taatcctttt ctacatcaga
     3181 taacttcggt ttgttatcat cgtctgtatt gtcatcaatt ggcgcgcctc tccctgcagg
     3241 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3301 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3361 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3661 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3901 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4081 tgtttaaacc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4681 cgatggttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5221 aagtgccacc tgacgtcgaa agaacgcggc attagcaaca tttgcgaagc gaacgatttt
     5281 gcgaactgct attaattaac tctggcgcgc cactactttt tggaactctt gttgttcttt
     5341 ggagttcaat gcgtccatct ttacagtcct gtcttattgt tcttgatttg tgccccgtaa
     5401 aatactgtta cttggttctg gcgaggtatt ggatagttcc tttttataaa ggccatgaag
     5461 gtttttcttt ccaatttttt ttttttcgtc attatagaaa tcattacgac cgagattccc
     5521 ggcatatgcg ttttcgacac tggatggcgg cgttagtatc gaatcgacag cagtatagcg
     5581 accagcattc acatacgatt gacgcatgat attactttct gcgcacttaa cttcgcatct
     5641 gggcagatga tgtcgaggcg aaaaaaaata taaatcacgc taacatttga ttaaaataga
     5701 acaactacaa tataaaaaaa ctatacaaat gacaagttct tgaaaacaag aatcttttta
     5761 ttgtcagtac tgattataat tggccagtct ttttcaaata agcattccaa ccagcatctc
     5821 tatacctttt accttcaata tctggatctc ttcctttacc aaacaatcct ctaccaacaa
     5881 tgataatatc agttccagtg ctaacaactt catcaacagt tctatattgt tgtcctaatc
     5941 catcaccttt atcatctaat ccaactccag gtgtcataat aagccaatca aatccttctt
     6001 cttgtccacc catatcacgt tgggcaataa atccaataac aaattcctta tcggatttag
     6061 caatttcaac agttttttga gaatattctc cataagctaa tgatcccact gatgataatt
     6121 cagctaacat caataaccct cttggctctt ggttggtggt ggtttcttta gctccctgtt
     6181 ttaatccttc aactactcca ttcccagtga caccatgagc attagtaata tctgcccaac
     6241 tactaatttt ataaactcca ccaatatatt gtttcttcac ggtattacca atatcagcaa
     6301 attttctatc ttcaaaaatc ataaattgat gtttacgtga aagttctaat aatggttcaa
     6361 tagtggattc ataggaaaaa tcattgatta tatcaatatg agtcttgatt aagcatacat
     6421 aaggacccaa tttatcaatt aattcaagaa attccttagt ggtatcaaca tcaattgatg
     6481 cacataaatt ggttttcttc agttccatta atcgaaataa tcgttgtgct actggtgagg
     6541 catgagtttc tgctctctca ctataggtct tagtgttgac tgtcatggtt gtttatgttc
     6601 ggatgtgatg tgagaactgt atcctagcaa gattttaaaa ggaagtatat gaaagaagaa
     6661 cctcagtggc aaatcctaac cttttatatt tctctacagg ggcgcggcgt ggggacaatt
     6721 caacgcgtct gtgaggggag cgtttccctg ctcgcaggtc cgcagcgagg agccgtaatt
     6781 tttgcttcgc gccgtgcggc catcaaaatg tatggatgca aatgattata catggggatg
     6841 tatgggctaa atgtacgggc gacagtcaca tcatgcccct gagctgcgca cgtcaagact
     6901 gtcaaggagg gtattctggg cctccatgtc gctggccggg tgacccggcg gggacgaggc
     6961 aagctaaacc gatcggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     7021 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
     7081 tggagaaaca gaataatata taaacctgt
//
