LOCUS       sequence-313196-        8380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert: Pact1-lexA-ER-B112-phosphodegron-tCyc1. The transcription 
            factor LexA-ER-B112 (see addgene FRP880) is degraded upon addition 
            of alpha factor. Backbone: pRG201 (addGene). Marker: Met15..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8380)
  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 8380)
  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: pCL21-decoder
FEATURES             Location/Qualifiers
     source          1..8380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(168..756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(248..267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(935..1726,1727..1795))
                     /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     1558..1577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1796..1900)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2041..2391
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             2392..3726
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     primer_bind     4399..4421
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4413..4430
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4414..4430
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4437..4456
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4437..4455
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5145..5396,5397..5750)
                     /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     5688..5709
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             5769..6704
                     /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             5817..6707
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      7333..7582
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7349..7367)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(7745..7763)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7784..7800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7784..7800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7797..7819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7808..7824
                     /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(join(8377..8380,1..14))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
       61 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttgttta
      781 aacctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatgg
     1381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1921 cacmtgacgt cgaaagaacg cggcattagc aacatttgcg aagcgaacga ttttgcgaac
     1981 tgctattaat taaacacaca ccataccttc ccatatgtcc tggcgcgccc ttagctctca
     2041 ttattttttg ctttttctct tgaggtcaca tgatcgcaaa atggcaaatg gcacgtgaag
     2101 ctgtcgatat tggggaactg tggtggttgg caaatgacta attaagttag tcaaggcgcc
     2161 atcctcatga aaactgtgta acataataac cgaagtgtcg aaaaggtggc accttgtcca
     2221 attgaacacg ctcgatgaaa aaaataagat atatataagg ttaagtaaag cgtctgttag
     2281 aaaggaagtt tttccttttt cttgctctct tgtcttttca tctactattt ccttcgtgta
     2341 atacagggtc gtcagataca tagatacaat tctattaccc ccatccatac aatgccatct
     2401 catttcgata ctgttcaact acacgccggt caagagaacc ctggtgacaa tgctcacaga
     2461 tccagagctg taccaattta cgccaccact tcttatgttt tcgaaaactc taagcatggt
     2521 tcgcaattgt ttggtttaga agttccaggt tacgtctatt cccgtttcca aaacccaacc
     2581 agtaatgttt tggaagaaag aattgctgct ttagaaggtg gtgctgctgc tttggctgtt
     2641 tcctccggtc aagccgctca aacccttgcc atccaaggtt tggcacacac tggtgacaac
     2701 atcgtttcca cttcttactt atacggtggt acttataacc agttcaaaat ctcgttcaaa
     2761 agatttggta tcgaggctag atttgttgaa ggtgacaatc cagaagaatt tgaaaaggtc
     2821 tttgatgaaa gaaccaaggc tgtttatttg gaaaccattg gtaatccaaa gtacaatgtt
     2881 ccggattttg aaaaaattgt tgcaattgct cacaaacacg gtattccagt tgtcgttgac
     2941 aacacatttg gtgccggtgg ttacttctgt cagccaatta aatacggtgc tgatattgta
     3001 acacattctg ctaccaaatg gattggtggt catggtacta ctatcggtgg tattattgtt
     3061 gactctggta agttcccatg gaaggactac ccagaaaagt tccctcaatt ctctcaacct
     3121 gccgaaggtt atcacggtac tatctacaat gaagcctacg gtaacttggc atacatcgtt
     3181 catgttagaa ctgaactatt aagagatttg ggtccattga tgaacccatt tgcctctttc
     3241 ttgctactac aaggtgttga aacattatct ttgagagctg aaagacacgg tgaaaatgca
     3301 ttgaagttag ccaaatggtt agaacaatcc ccatacgtat cttgggtttc ataccctggt
     3361 ttagcatctc attctcatca tgaaaatgct aagaagtatc tatctaacgg tttcggtggt
     3421 gtcttatctt tcggtgtaaa agacttacca aatgccgaca aggaaactga cccattcaaa
     3481 ctttctggtg ctcaagttgt tgacaattta aagctagcct ctaacttggc caatgttggt
     3541 gatgccaaga ccttagtcat tgctccatac ttcactaccc acaaacaatt aaatgacaaa
     3601 gaaaagttgg catctggtgt taccaaggac ttaattcgtg tctctgttgg tatcgaattt
     3661 attgatgaca ttattgcaga cttccagcaa tcttttgaaa ctgttttcgc tggccaaaaa
     3721 ccatgagtgt gcgtaatgag ttgtaaaatt atgtataaac ctactttctc tcacaagtac
     3781 tatactttta taaaacgaac tttattgaaa tgaatatcct ttttttccct tgttacatgt
     3841 cgtgactcgt actttgaacc taaattgttc taacatcaaa gaacagtgtt aattcgcagt
     3901 cgagaagaaa aatatggtga acaagactca tctacttcat gagactactt tacgcctcct
     3961 ataaagctgt cacactggat aaatttattg taggaccaag ttacaaaaga ggatgatgga
     4021 ggtttcttta caataaagaa gcacatgtgt gttaacgttt ttagtatttg cttgttatgt
     4081 aaatcaggaa aacttcgcgg gatttggttg gatgctactt tccatacaat aaatattata
     4141 gatcaaaaaa gccaaattac aagtaaagat tagtaaagct gttgcaattc catcgttgat
     4201 aaaaatgtta gttattaaat ataaaagtca gaataggtga acttggattt aattgttggc
     4261 atttcgttgc tgctagaggc cataatatta gatagccagg acatactagg tctcctcgtg
     4321 gtataggaat ccataaaatg gaattggtga cgatcggatg tgctgcaagg cgattaagtt
     4381 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
     4441 acgactcact atagggcgaa ttggagaaac agaataatat ataaacctgt ataatataac
     4501 cttgaagact atatttcttt tcgcggccgc ctatagggcg aattggagct ccaccgcggt
     4561 ggcggccgct ctagagaagc gggtaagctg ccacagcaat taatgcacaa catttaacct
     4621 acattcttcc ttatcggatc ctcaaaaccc ttaaaaacat atgcctcacc ctaacatatt
     4681 ttccaattaa ccctcaatat ttctctgtca cccggcctct attttccatt ttcttcttta
     4741 cccgccacgc gtttttttct ttcaaatttt tttcttcctt cttctttttc ttccacgtcc
     4801 tcttgcataa ataaataaac cgttttgaaa ccaaactcgc ctctctctct cctttttgaa
     4861 atatttttgg gtttgtttga tcctttcctt cccaatctct cttgtttaat atatattcat
     4921 ttatatcacg ctctcttttt atcttccttt ttttcctctc tcttgtattc ttccttcccc
     4981 tttctactca aaccaagaag aaaaagaaaa ggtcaatctt tgttaaagaa taggatcttc
     5041 tactacatca gcttttagat ttttcacgct tactgctttt ttcttcccaa gatcgaaaat
     5101 ttactgaatt aacaactagt ggatcccccg ggctgcagga attcatgaaa gcgttaacgg
     5161 ccaggcaaca agaggtgttt gatctcatcc gtgatcacat cagccagaca ggtatgccgc
     5221 cgacgcgtgc ggaaatcgcg cagcgtttgg ggttccgttc cccaaacgcg gctgaagaac
     5281 atctgaaggc gctggcacgc aaaggcgtta ttgaaattgt ttccggcgca tcacgcggga
     5341 ttcgtctgtt gcaggaagag gaagaagggt tgccgctggt aggtcgtgtg gctgccggtg
     5401 aaccacttct ggcgcaacag catattgaag gtcattatca ggtcgatcct tccttattca
     5461 agccgaatgc tgatttcctg ctgcgcgtca gcgggatgtc gatgaaagac atcggcatta
     5521 tggatggtga cttgctggca gtgcataaaa ctcaggatgt acgtaacggt caggtcgttg
     5581 tcgcacgtat tgatgacgag gttaccgtta agcgcctgaa aaaacagggc aataaagtcg
     5641 aactgttgcc agaaaatagc gagtttaaac caattgtcgt agatcttcgt cagcagagct
     5701 tcaccattga agggctggcg gttggggtta ttcgcaacgg cgactggctg ggttctaagg
     5761 atatctctgc tggagacatg agagctgcca acctttggcc aagcccgctc atgatcaaac
     5821 gctctaagaa gaacagcctg gccttgtccc tgacggccga ccagatggtc agtgccttgt
     5881 tggatgctga gccccccata ctctattccg agtatgatcc taccagaccc ttcagtgaag
     5941 cttcgatgat gggcttactg accaacctgg cagacaggga gctggttcac atgatcaact
     6001 gggcgaagag ggtgccaggc tttgtggatt tgaccctcca tgatcaggtc caccttctag
     6061 aatgtgcctg gttagagatc ctgatgattg gtctcgtctg gcgctccatg gagcacccag
     6121 tgaagctact gtttgctcct aacttgctct tggacaggaa ccagggaaaa tgtgtagagg
     6181 gcatggtgga gatcttcgac atgctgctgg ctacatcatc tcggttccgc atgatgaatc
     6241 tgcagggaga ggagtttgtg tgcctcaaat ctattatttt gcttaattct ggagtgtaca
     6301 catttctgtc cagcaccctg aagtctctgg aagagaagga ccatatccac cgagtcctgg
     6361 acaagatcac agacactttg atccacctga tggccaaggc aggcctgacc ctgcagcagc
     6421 agcaccagcg gctggcccag ctcctcctca tcctctccca catcaggcac atgagtaaca
     6481 aaggcatgga gcatctgtac agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc
     6541 tgctggagat gctggacgcc caccgcctac atgcgcccac tagccgtgga ggggcatccg
     6601 tggaggagac ggaccaaagc cacttggcca ctgcgggctc tacttcatcg cattccttgc
     6661 aaaagtatta catcacgggg gaggcagagg gtttccctgc cacagtcgat atcatgcagg
     6721 atctgccggg caacgataac agcaccgcgg gcgaatttcc aggtattact ttgagaatcc
     6781 aagaaactga tatgttgtat aaaggtgata ctttgtattt ggattggttg gaagatggta
     6841 ttgctgaatt ggttttcgat gctccaggtt ctgttaataa attggatact gctgttgctt
     6901 ctttgggtga agctattggt gttttggaac aacaatctga tttgatttgg gaaactttga
     6961 ctgttaaaga tgctaaagtt aatttcgatt ctggtttgga aaaattcgaa gaagctattc
     7021 catctgctga tgatttcgat ccagttgctg aaagaagatc ctctggtgaa tttagagctg
     7081 aaagacattc tggtggtact gatttgtgtt tcaagcttgg aagcgattca aaaaggggca
     7141 atatccctaa accgttaaac ttgagcaaac ccatccctag agagtttagt ggtttactta
     7201 cacccatctc cgcgtccaat gaaaaaaaaa ttgagaactt tattaaaacg aatgctgcgt
     7261 cccaggcaaa aactcctttg taaacgcgtt taaagcttat cctcgagaca ggcccctttt
     7321 cctttgtcta tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat
     7381 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     7441 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     7501 gtacaaacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     7561 acgctcgaag gctttaattt gcaaggttcg cagtttacac tctcatcgtc gctctcatca
     7621 tcgcttccgt tgttgttttc cttagtagcg tctgcttggt acccagcttc ggccgtattt
     7681 acagataacg atgtcgttat tatatatata tatatatatt gtcacttgta cccagctttt
     7741 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
     7801 tgtgaaattg ttatccgctc acaacagctg tcagatatag tcggattgcc cttttaagca
     7861 atcaatagtg ttttatttgc aacaatgtcg tcatagttta atatgtccta taagatgtta
     7921 acttgctcaa cattcaacaa agtttggtct cttggcccag ttggttaagg caccgtgcta
     7981 ataacgcggg gatcagcggt tcgatcccgc tagagaccat ttatttttaa attcgaccgt
     8041 ctgacagacg gtgcacttat tacttaattt tttttttttt ttaatcctcg ttaaactgag
     8101 tacataatca gcaacaatat aatataaaag ttttgctttt ttatgtacgt acaatagggg
     8161 tacttaaata aataaataaa caataaaaca atcaataaat aaataatata cgatcataaa
     8221 ttacgagata tatttcatag tcttgacttc tccagccgaa cgtcattaaa tttaaggcag
     8281 aaaagataaa caagggcgcg ccagagcctg caggcattaa tgaatcggcc aacgcgcggg
     8341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
//
