LOCUS       sequence-90259-e        7244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LexA-ER-B42 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7244)
  AUTHORS   Ottoz DS, Rudolf F, Stelling J
  TITLE     Inducible, tightly regulated and growth condition-independent 
            transcription factor in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2014 Jul 17. pii: gku616.
  PUBMED    25034689
REFERENCE   2  (bases 1 to 7244)
  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: "Nucleic 
            Acids Res. 2014 Jul 17. pii: gku616."
COMMENT     Sequence Label: FRP718_PACT1(-1-520)-LexA-ER-haB42-TCYC1
FEATURES             Location/Qualifiers
     source          1..7244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             509..1168
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1429..1884)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1566..1587)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1778..1797
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2014..2036
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2028..2045
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2029..2045
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2052..2071
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2052..2070
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2709..2960,2961..3314)
                     /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     3252..3273
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             3333..4268
                     /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             3381..4271
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     CDS             4320..4556
                     /codon_start=1
                     /product="yeast transcriptional activator cretaed from E. 
                     coli genomic DNA fragments (Ma and Ptashne, 1987)"
                     /label=B42 transcriptional activator
                     /translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI
                     LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD"
     primer_bind     complement(4589..4605)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4590..4606)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4633..4882
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4649..4667)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(4994..5012)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5033..5049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5033..5049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5046..5068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5057..5073
                     /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)."
     promoter        complement(join(5081..5087,5088..5105,5106..5111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5264..5281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5435..6023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6194..6985,6986..7054))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6817..6836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7055..7159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7217..7235
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta tcgtttgaac
      241 acggcattag tcagggaagt cataacacag tcctttcccg caattttctt tttctattac
      301 tcttggcctc ctctagtaca ctctatattt ttttatgcct cttaattaag gtaatgattt
      361 tcattttttt ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc
      421 acataatgaa ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa
      481 tgagcaggca agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat
      541 tacaaatgaa accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga
      601 gcactcgatc ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca
      661 agtgattaac gtccacacag gtatagggtt tctggaccac atgatacatg ctctggccaa
      721 gcattccggc tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca
      781 caccactgaa gactgcggga ttgctctcgg tcaggctttt aaagaggccc tactggcgcg
      841 tggagtaaaa aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt
      901 ggtagatctt tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt
      961 aggagatctc tcttgcgaga tgatcccgca ttttcttgaa agttttgcag aggctagcag
     1021 aattaccctc cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc
     1081 gttcaaggct cttgcggttg ccataagaga agccacctcg cccaatggaa ccaacgatgt
     1141 tccctccacc aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga
     1201 tatatataca tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt
     1261 atgatactga agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc
     1321 gctttccttt tttctttttg ctttttcttt ttttttctct tgaactcgac ggatcatatg
     1381 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg
     1441 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1501 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1561 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1621 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1681 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1741 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1801 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1861 ttaatgcgcc gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg
     1921 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg
     1981 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2041 ccagtgaatt gtaatacgac tcactatagg gcgaattgga gaaacagcgg ccgcctatag
     2101 ggcgaattgg agctccaccg cggtggcggc cgctctagag aagcgggtaa gctgccacag
     2161 caattaatgc acaacattta acctacattc ttccttatcg gatcctcaaa acccttaaaa
     2221 acatatgcct caccctaaca tattttccaa ttaaccctca atatttctct gtcacccggc
     2281 ctctattttc cattttcttc tttacccgcc acgcgttttt ttctttcaaa tttttttctt
     2341 ccttcttctt tttcttccac gtcctcttgc ataaataaat aaaccgtttt gaaaccaaac
     2401 tcgcctctct ctctcctttt tgaaatattt ttgggtttgt ttgatccttt ccttcccaat
     2461 ctctcttgtt taatatatat tcatttatat cacgctctct ttttatcttc ctttttttcc
     2521 tctctcttgt attcttcctt cccctttcta ctcaaaccaa gaagaaaaag aaaaggtcaa
     2581 tctttgttaa agaataggat cttctactac atcagctttt agatttttca cgcttactgc
     2641 ttttttcttc ccaagatcga aaatttactg aattaacaac tagtggatcc cccgggctgc
     2701 aggaattcat gaaagcgtta acggccaggc aacaagaggt gtttgatctc atccgtgatc
     2761 acatcagcca gacaggtatg ccgccgacgc gtgcggaaat cgcgcagcgt ttggggttcc
     2821 gttccccaaa cgcggctgaa gaacatctga aggcgctggc acgcaaaggc gttattgaaa
     2881 ttgtttccgg cgcatcacgc gggattcgtc tgttgcagga agaggaagaa gggttgccgc
     2941 tggtaggtcg tgtggctgcc ggtgaaccac ttctggcgca acagcatatt gaaggtcatt
     3001 atcaggtcga tccttcctta ttcaagccga atgctgattt cctgctgcgc gtcagcggga
     3061 tgtcgatgaa agacatcggc attatggatg gtgacttgct ggcagtgcat aaaactcagg
     3121 atgtacgtaa cggtcaggtc gttgtcgcac gtattgatga cgaggttacc gttaagcgcc
     3181 tgaaaaaaca gggcaataaa gtcgaactgt tgccagaaaa tagcgagttt aaaccaattg
     3241 tcgtagatct tcgtcagcag agcttcacca ttgaagggct ggcggttggg gttattcgca
     3301 acggcgactg gctgggttct aaggatatct ctgctggaga catgagagct gccaaccttt
     3361 ggccaagccc gctcatgatc aaacgctcta agaagaacag cctggccttg tccctgacgg
     3421 ccgaccagat ggtcagtgcc ttgttggatg ctgagccccc catactctat tccgagtatg
     3481 atcctaccag acccttcagt gaagcttcga tgatgggctt actgaccaac ctggcagaca
     3541 gggagctggt tcacatgatc aactgggcga agagggtgcc aggctttgtg gatttgaccc
     3601 tccatgatca ggtccacctt ctagaatgtg cctggttaga gatcctgatg attggtctcg
     3661 tctggcgctc catggagcac ccagtgaagc tactgtttgc tcctaacttg ctcttggaca
     3721 ggaaccaggg aaaatgtgta gagggcatgg tggagatctt cgacatgctg ctggctacat
     3781 catctcggtt ccgcatgatg aatctgcagg gagaggagtt tgtgtgcctc aaatctatta
     3841 ttttgcttaa ttctggagtg tacacatttc tgtccagcac cctgaagtct ctggaagaga
     3901 aggaccatat ccaccgagtc ctggacaaga tcacagacac tttgatccac ctgatggcca
     3961 aggcaggcct gaccctgcag cagcagcacc agcggctggc ccagctcctc ctcatcctct
     4021 cccacatcag gcacatgagt aacaaaggca tggagcatct gtacagcatg aagtgcaaga
     4081 acgtggtgcc cctctatgac ctgctgctgg agatgctgga cgcccaccgc ctacatgcgc
     4141 ccactagccg tggaggggca tccgtggagg agacggacca aagccacttg gccactgcgg
     4201 gctctacttc atcgcattcc ttgcaaaagt attacatcac gggggaggca gagggtttcc
     4261 ctgccacagt cgatatcatg caggatctgc cgggcaacga taacagcacc gcgggcggta
     4321 tcaataaaga catcgaggag tgcaatgcca tcattgagca gtttatcgac tacctgcgca
     4381 ccggacagga gatgccgatg gaaatggcgg atcaggcgat taacgtggtg ccgggcatga
     4441 cgccgaaaac cattcttcac gccgggccgc cgatccagcc tgactggctg aaatcgaatg
     4501 gttttcatga aattgaagcg gatgttaacg ataccagcct cttgctgagt ggagatgcct
     4561 ccaagcttta aacgcgttta aagcttatcg ataccgtcga cctcgagaca ggcccctttt
     4621 cctttgtcta tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat
     4681 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     4741 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     4801 gtacaaacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     4861 acgctcgaag gctttaattt gcaaggttcg cagtttacac tctcatcgtc gctctcatca
     4921 tcgcttccgt tgttgttttc cttagtagcg tctgcttggt acccagcttc ggccgtgtac
     4981 ccagcttttg ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc
     5041 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatagga gccggaagca
     5101 taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt gcgttgcgct
     5161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctcggc ccccctgacg
     5461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5521 accaggcgtt cccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     5641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6241 ttcgttcatc catagttgcc tgactgcccg tcgtgtagat aactacgata cgggagggct
     6301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6601 tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc ttattatcat
     7201 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
