LOCUS       sequence-108558-        7398 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 7398)
  AUTHORS   Gnugge R, Liphardt T, Rudolf F
  TITLE     A shuttle vector series for precise genetic engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2015 Dec 9. doi: 10.1002/yea.3144.
  PUBMED    26647923
REFERENCE   2  (bases 1 to 7398)
  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: "Yeast. 2015
            Dec 9. doi: 10.1002/yea.3144."
COMMENT     Sequence Label: FRP1625
FEATURES             Location/Qualifiers
     source          1..7398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(354..1448)
                     /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"
     promoter        complement(1449..1854)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     2050..2072
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2064..2081
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2065..2081
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2088..2107
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2088..2106
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2796..3047,3048..3401)
                     /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     3339..3360
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             3420..4355
                     /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             3468..4358
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     CDS             4407..4643
                     /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(4676..4692)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4677..4693)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4720..4969
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4736..4754)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(5132..5150)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5171..5187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5171..5187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5184..5206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5195..5211
                     /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(5470..5487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5641..6229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5721..5740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6408..7199,7200..7268))
                     /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     7031..7050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7269..7373)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc taactttttg tgtggtgccc tcctccttgt caatattaat
      121 gttaaagtgc aattcttttt ccttatcacg ttgagccatt agtatcaatt tgcttacctg
      181 tattccttta catcctcctt tttctccttc ttgataaatg tatgtagatt gcgtatatag
      241 tttcgtctac cctatgaaca tattccattt tgtaatttcg tgtcgcatat gtttctatta
      301 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
      361 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
      421 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
      481 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
      541 gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
      601 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
      661 aaggaacctg ggataacgga ggcttcatcg gagatgatgt caccaaacat gttgctggtg
      721 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
      781 aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc cacagttttt
      841 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
      901 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
      961 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     1021 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     1081 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     1141 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actaccagta
     1201 ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc
     1261 agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga
     1321 ttttggaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     1381 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     1441 gcagacatta gaatggtata tccttgaaat atatatatat attgctgaaa tgtaaaaggt
     1501 aagaaaagtt agaaagtaag acgattgcta accacctatt ggaaaaaaca ataggtcctt
     1561 aaataatatt gtcaacttca agtattgtga tgcaagcatt tagtcatgaa cgcttctcta
     1621 ttctatatga aaagccggtt ccggcgctct cacctttcct ttttctccca atttttcagt
     1681 tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa
     1741 tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg
     1801 ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac agttaactgc
     1861 ggtcaagata tttcttgaat caggcgcctt agaccgctcg gccaaacaac caattacttg
     1921 ttgagaaata gagtataatt atcctataaa tataacgttt ttgaacacac atgaacaagg
     1981 aagtacagga caattgattt tcgatcggat gtgctgcaag gcgattaagt tgggtaacgc
     2041 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
     2101 tatagggcga attggagaaa cagaataata tataaacctg tataatataa ccttgaagac
     2161 tatatttctt ttcgcggccg cctatagggc gaattggagc tccaccgcgg tggcggccgc
     2221 tctagagaag cgggtaagct gccacagcaa ttaatgcaca acatttaacc tacattcttc
     2281 cttatcggat cctcaaaacc cttaaaaaca tatgcctcac cctaacatat tttccaatta
     2341 accctcaata tttctctgtc acccggcctc tattttccat tttcttcttt acccgccacg
     2401 cgtttttttc tttcaaattt ttttcttcct tcttcttttt cttccacgtc ctcttgcata
     2461 aataaataaa ccgttttgaa accaaactcg cctctctctc tcctttttga aatatttttg
     2521 ggtttgtttg atcctttcct tcccaatctc tcttgtttaa tatatattca tttatatcac
     2581 gctctctttt tatcttcctt tttttcctct ctcttgtatt cttccttccc ctttctactc
     2641 aaaccaagaa gaaaaagaaa aggtcaatct ttgttaaaga ataggatctt ctactacatc
     2701 agcttttaga tttttcacgc ttactgcttt tttcttccca agatcgaaaa tttactgaat
     2761 taacaactag tggatccccc gggctgcagg aattcatgaa agcgttaacg gccaggcaac
     2821 aagaggtgtt tgatctcatc cgtgatcaca tcagccagac aggtatgccg ccgacgcgtg
     2881 cggaaatcgc gcagcgtttg gggttccgtt ccccaaacgc ggctgaagaa catctgaagg
     2941 cgctggcacg caaaggcgtt attgaaattg tttccggcgc atcacgcggg attcgtctgt
     3001 tgcaggaaga ggaagaaggg ttgccgctgg taggtcgtgt ggctgccggt gaaccacttc
     3061 tggcgcaaca gcatattgaa ggtcattatc aggtcgatcc ttccttattc aagccgaatg
     3121 ctgatttcct gctgcgcgtc agcgggatgt cgatgaaaga catcggcatt atggatggtg
     3181 acttgctggc agtgcataaa actcaggatg tacgtaacgg tcaggtcgtt gtcgcacgta
     3241 ttgatgacga ggttaccgtt aagcgcctga aaaaacaggg caataaagtc gaactgttgc
     3301 cagaaaatag cgagtttaaa ccaattgtcg tagatcttcg tcagcagagc ttcaccattg
     3361 aagggctggc ggttggggtt attcgcaacg gcgactggct gggttctaag gatatctctg
     3421 ctggagacat gagagctgcc aacctttggc caagcccgct catgatcaaa cgctctaaga
     3481 agaacagcct ggccttgtcc ctgacggccg accagatggt cagtgccttg ttggatgctg
     3541 agccccccat actctattcc gagtatgatc ctaccagacc cttcagtgaa gcttcgatga
     3601 tgggcttact gaccaacctg gcagacaggg agctggttca catgatcaac tgggcgaaga
     3661 gggtgccagg ctttgtggat ttgaccctcc atgatcaggt ccaccttcta gaatgtgcct
     3721 ggttagagat cctgatgatt ggtctcgtct ggcgctccat ggagcaccca gtgaagctac
     3781 tgtttgctcc taacttgctc ttggacagga accagggaaa atgtgtagag ggcatggtgg
     3841 agatcttcga catgctgctg gctacatcat ctcggttccg catgatgaat ctgcagggag
     3901 aggagtttgt gtgcctcaaa tctattattt tgcttaattc tggagtgtac acatttctgt
     3961 ccagcaccct gaagtctctg gaagagaagg accatatcca ccgagtcctg gacaagatca
     4021 cagacacttt gatccacctg atggccaagg caggcctgac cctgcagcag cagcaccagc
     4081 ggctggccca gctcctcctc atcctctccc acatcaggca catgagtaac aaaggcatgg
     4141 agcatctgta cagcatgaag tgcaagaacg tggtgcccct ctatgacctg ctgctggaga
     4201 tgctggacgc ccaccgccta catgcgccca ctagccgtgg aggggcatcc gtggaggaga
     4261 cggaccaaag ccacttggcc actgcgggct ctacttcatc gcattccttg caaaagtatt
     4321 acatcacggg ggaggcagag ggtttccctg ccacagtcga tatcatgcag gatctgccgg
     4381 gcaacgataa cagcaccgcg ggcggtatca ataaagacat cgaggagtgc aatgccatca
     4441 ttgagcagtt tatcgactac ctgcgcaccg gacaggagat gccgatggaa atggcggatc
     4501 aggcgattaa cgtggtgccg ggcatgacgc cgaaaaccat tcttcacgcc gggccgccga
     4561 tccagcctga ctggctgaaa tcgaatggtt ttcatgaaat tgaagcggat gttaacgata
     4621 ccagcctctt gctgagtgga gatgcctcca agctttaaac gcgtttaaag cttatcgata
     4681 ccgtcgacct cgagacaggc cccttttcct ttgtctatat catgtaatta gttatgtcac
     4741 gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac
     4801 ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt
     4861 tatatttcaa atttttcttt tttttctgta caaacgcgtg tacgcatgta acattatact
     4921 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca aggttcgcag
     4981 tttacactct catcgtcgct ctcatcatcg cttccgttgt tgttttcctt agtagcgtct
     5041 gcttggtacc cagcttcggc cgtatttaca gataacgatg tcgttattat atatatatat
     5101 atatattgtc acttgtaccc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     5161 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca acagctgtaa
     5221 tatatagtag taacctgaaa aatagtcaat gcccgtggtt catttgccaa tagaacagtc
     5281 gatgtgatca atgtacacga aagtcaatat aaaatatcca aatagccaat ggtgtcagga
     5341 cggctaaggc aaatgcaact ttgaaatact tattcggttt gcatcttggc gcgcctcttc
     5401 ctgcaggcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5461 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5521 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5581 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5641 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5701 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5761 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5821 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5881 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5941 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6001 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6061 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6121 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6181 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6241 gatcttttgt ttaaacctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6361 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6421 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6481 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6541 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6601 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6661 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6721 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6781 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6841 ggtcctccga tggttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6901 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6961 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7021 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7081 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7141 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7201 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7261 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7321 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7381 ccccgaaaag tgccacct
//
