LOCUS       sequence-180366-        9131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses estradiol-inducible synthetic transcription factor in 
            yeast under control of GAL1 promoter. It activates transcription of 
            promoters containing Zif268 binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9131)
  AUTHORS   Aranda-Diaz A, Mace K, Zuleta I, Harrigan P, El-Samad H
  TITLE     Robust Synthetic Circuits for Two-Dimensional Control of Gene 
            Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251.
  PUBMED    27930885
REFERENCE   2  (bases 1 to 9131)
  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: "ACS Synth 
            Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251."
COMMENT     Sequence Label: pHES873
FEATURES             Location/Qualifiers
     source          1..9131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..666
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     630..653
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     promoter        complement(3670..3688)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3709..3725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3709..3725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3722..3744)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3733..3749
                     /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(3757..3763,3764..3781,3782..3787))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3802..3823
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3940..3957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4111..4699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4191..4210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4870..5661,5662..5730))
                     /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     5493..5512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5731..5835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5920..5938
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5976..5998)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6098..6117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6473..6928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6610..6631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6822..6841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7054..7076
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7068..7085
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7069..7085
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7092..7111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7092..7110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7677..8020
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      8839..9036
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac
       61 atggcattgc caccatatac atatccatat acatatccat atctaatctt acttatatgt
      121 tgtggaaatg taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc
      181 agtaatacgc ttaactgctc attgctatat tgaagtacgg actagaagcc gccgagcggg
      241 cgacagccct ccgacggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc
      301 tgaaacgcag atgtgcctca cgccacactg ctccgaacaa taaagattct acaatactag
      361 cttttatagt tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta
      421 acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg
      481 ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc
      541 tgcataacca ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa
      601 tgtcataaaa gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga
      661 aaaaactata gctagcaata aaatgggtac ccgcccatat gcttgccctg tcgagtcctg
      721 cgatcgccgc ttttctcgct cggatgagct tacccgccat atccgcatcc ataccggtca
      781 gaagcccttc cagtgtcgaa tctgcatgcg taacttcagt cgtagtgacc accttaccac
      841 ccacatccgc acccacacag gcgagaagcc ttttgcctgt gacatttgtg ggaggaagtt
      901 tgccaggagt gatgaacgca agaggcatac caaaatccat acaggtagat ccggtgacgg
      961 tgctggttta aagttcaata aagtcagagt tgtgagagca ctggatgctg ttgctctccc
     1021 acagccattg ggcgttccaa atgaaagcca agccctaagc cagagattca ctttttcacc
     1081 aggtcaagac atacagttga ttccaccact gatcaacctg ttaatgagca ttgaaccaga
     1141 tgtgatctat gcaggacatg acaacacaaa acctgacacc tccagttctt tgctgacaag
     1201 tcttaatcaa ctaggcgaga ggcaacttct ttcagtagtc aagtggtcta aatcattgcc
     1261 aggttttcga aacttacata ttgatgacca gataactctc attcagtatt cttggatgag
     1321 cttaatggtg tttggtctag gatggagatc ctacaaacat gtcagtgggc agatgctgta
     1381 ttttgcacct gatctaatac taaatgaaca gcggatgaaa gaatcatcat tctattcatt
     1441 atgccttacc atgtggcaga tcccacagga gtttgtcaag cttcaagtta gccaagaaga
     1501 gttcctctgt atgaaagtat tgttacttct taatacaatt cctttggaag ggctacgaag
     1561 tcaaacccag tttgaggaga tgaggtcaag ctacattaga gagctcatca aggcaattgg
     1621 tttgaggcaa aaaggagttg tgtcgagctc acagcgtttc tatcaactta caaaacttct
     1681 tgataactcg catgatcttg tcaaacagct tcatctgtac tgcttgaata catttatcca
     1741 gtcccgggca ctgagtgttg aatttccaga aatgatgtct gaagttattg ctgcacaatt
     1801 acccaagata ttggcaggga tggtgaaacc ccttctcttt cataaaaagg cggccgcagg
     1861 tgacggtgct ggtttaatta acatgacggt cgaccatgat ttcaatagcg aagatatttt
     1921 attccccata gaaagcatga gtagtataca atacgtggag aataataacc caaataatat
     1981 taacaacgat gttatcccgt attctctaga tatcaaaaac actgtcttag atagtgcgga
     2041 tctcaatgac attcaaaatc aagaaacttc actgaatttg gggcttcctc cactatcttt
     2101 cgactctcca ctgcccgtaa cggaaacgat accatccact accgataaca gcttgcattt
     2161 gaaagctgat agcaacaaaa atcgcgatgc aagaactatt gaaaatgata gtgaaattaa
     2221 gagtactaat aatgctagtg gctctggggc aaatcaatac acaactctta cttcacctta
     2281 tcctatgaac gacattttgt acaacatgaa caatccgtta caatcaccgt caccttcatc
     2341 ggtacctcaa aatccgacta taaatcctcc cataaataca gcaagtaacg aaactaattt
     2401 atcgcctcaa acttcaaatg gtaatgaaac tcttatatct cctcgagccc aacaacatac
     2461 gtccattaaa gataatcgtc tgtccttacc taatggtgct aattcgaatc ttttcattga
     2521 cactaaccca aacaatttga acgaaaaact aagaaatcaa ttgaactcag atacaaattc
     2581 atattctaac tccatttcta attcaaactc caattctacg ggtaatttaa attccagtta
     2641 ttttaattca ctgaacatag actccatgct agatgattac gtttctagtg atctcttatt
     2701 gaatgatgat gatgatgaca ctaatttatc acgccgaaga tttagcgacg ttataacaaa
     2761 ccaagagctc taagcaaata gctaaattat atacgaatta atattatgat taagtgttta
     2821 cgtgagtgcg atatttttat tactatctta tacagttgta tatactctat aaaatgagtt
     2881 gtctattaat taacgcgatg aatgctttct gggtttacct ctccaacaac tctagtttac
     2941 ttctcaatac attcaattgt atttgatttg tcaatacttc atcattaatc aattctatag
     3001 ttttgttttt ctcgtttatt tccaaattta atgcatcaat tttattattc aatttgtcgt
     3061 tgattttggt taatgatttt atggtttgat ctctggcatt gattgtttgt gttagttttt
     3121 cattattgat aattaaatta tttaagttag ttatcaactc ggtgttttca agtttcaagt
     3181 tttcaatttc tttagagttt attagatttg tcaaagtttc tgaattgctt gattggtccg
     3241 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     3301 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     3361 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     3421 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     3481 tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat gtggtctcta
     3541 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     3601 tagagggtga acgttacaga aaagcaggct ggggtttaaa catttaaatg gccggcccag
     3661 cttttgttcc ctttagtgag ggttaattcc gagcttggcg taatcatggt catagctgtt
     3721 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3781 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3841 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3901 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcctcg gcgcgccgac
     5881 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5941 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6001 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6061 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6121 agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc
     6181 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     6241 agcacagact tagattggta tatatacgca tattgggtca gcagtacaga accgtcgacg
     6301 aagttgtaag tggtggatca gatatcatca ttgttggcag aggacttttc gccaagggta
     6361 gagatcctaa tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     6421 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     6481 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     6541 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     6601 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6661 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6721 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6781 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6841 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6901 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     6961 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     7021 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     7081 ccagtgaatt gtaatacgac tcactatagg gggccagcgt ggccgtttaa acagaacgaa
     7141 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     7201 attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa
     7261 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     7321 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     7381 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     7441 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     7501 tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata
     7561 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     7621 atgcacacgg tgtggtggtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca
     7681 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
     7741 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
     7801 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcggacc tgcgagcagg
     7861 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
     7921 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
     7981 taggatacag ttctcacatc acatccgaac ataaacaacc atgacagtca acactaagac
     8041 ctatagtgag agagcagaaa ctcatgcctc accagtagca caacgattat ttcgattaat
     8101 ggaactgaag aaaaccaatt tatgtgcatc aattgatgtt gataccacta aggaattcct
     8161 tgaattaatt gataaattgg gtccttatgt atgcttaatc aagactcata ttgatataat
     8221 caatgatttt tcctatgaat ccactattga accattatta gaactttcac gtaaacatca
     8281 atttatgatt tttgaagata gaaaatttgc tgatattggt aataccgtga agaaacaata
     8341 tattggtgga gtttataaaa ttagtagttg ggcagatatt actaatgctc atggtgtcac
     8401 tgggaatgga gtagttgaag gattaaaaca gggagctaaa gaaaccacca ccaaccaaga
     8461 gccaagaggg ttattgatgt tagctgaatt atcatcagtg ggatcattag catatggaga
     8521 atattctcaa aaaactgttg aaattgctaa atccgataag gaatttgtta ttggatttat
     8581 tgcccaacgt gatatgggtg gacaagaaga aggatttgat tggcttatta tgacacctgg
     8641 agttggatta gatgataaag gtgatggatt aggacaacaa tatagaactg ttgatgaagt
     8701 tgttagcact ggaactgata ttatcattgt tggtagagga ttgtttggta aaggaagaga
     8761 tccagatatt gaaggtaaaa ggtatagaga tgctggttgg aatgcttatt tgaaaaagac
     8821 tggccaatta taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     8881 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     8941 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     9001 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     9061 catccagtgt cgaaaacgac ctcgaattca tcgagtggat tcggctttgg gtaccgggcc
     9121 cttatattga a
//
