LOCUS       sequence-90258-e        7029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LexA-ER-VP16 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7029)
  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 7029)
  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: FRP467-PACT1(-1-520)-LexA-ER-haVP16
FEATURES             Location/Qualifiers
     source          1..7029
                     /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             4455..4688
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(4715..4731)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4716..4732)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4779..4797)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4818..4834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4818..4834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4831..4853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4842..4858
                     /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(4866..4872,4873..4890,4891..4896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5049..5066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5220..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5979..6770,6771..6839))
                     /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     6602..6621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6840..6944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7002..7020
                     /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 gcgggcgcgc
     4321 gtacgaaaaa caattacggg tctaccatcg agggcctgct cgatctcccg gacgacgacg
     4381 cccccgaaga ggcggggctg gcggctccgc gcctgtcctt tctccccgcg ggacacacgc
     4441 gcagactgtc gacggccccc ccgaccgatg tcagcctggg ggacgaattg cacttagacg
     4501 gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg
     4561 gggacgggga ttccccgggt ccgggattta ccccccacga ctccgccccc tacggcgctc
     4621 tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga attgacgagt
     4681 acggtgggaa gctttaaacg cgtttaaagc ttatcgatac cgtcgacctc gagggggggc
     4741 ccggtaccca gcttcggccg tgtacccagc ttttgttccc tttagtgagg gttaattccg
     4801 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4861 ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg
     4921 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4981 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5221 ttccataggc tcggcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5281 cgaaacccga caggactata aagataccag gcgttccccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact gcccgtcgtg
     6061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6361 aggcgagtta catgatcccc catgttgtga aaaaaagcgg ttagctcctt cggtcctccg
     6421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6961 gtgccacctg acgtcttatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7021 cctttcgtc
//
