LOCUS       sequence-180361-        9279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses progesterone-inducible synthetic 
            transcription factor in yeast. It activates transcription of 
            promoters containing Gal4 binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9279)
  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 9279)
  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: pHES941
FEATURES             Location/Qualifiers
     source          1..9279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    13..31
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    34..52
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             105..545
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECCYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLGMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        complement(3281..3299)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3320..3336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3320..3336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3333..3355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3344..3360
                     /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(3368..3374,3375..3392,3393..3398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3413..3434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3551..3568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3722..4310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3802..3821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4481..5272,5273..5341))
                     /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     5104..5123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5342..5446)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5531..5549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5587..5609)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5709..5728
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6084..6539)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6221..6242)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6433..6452
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6665..6687
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6679..6696
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6680..6696
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6703..6722
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6703..6721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7288..7631
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      8450..8647
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 aatagacctg aatccctatc agtgatagag atctccctat cagtgataga gattcttcct
       61 tgtttctttt tctgcaggtc gacaaacaaa tgctcgacat aaaaatgaag ctactgtctt
      121 ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac
      181 cgaagtgcgc caagtgtctg aagaacaact gggagtgttg ctactctccc aaaaccaaaa
      241 ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa agactggaac
      301 agctatttct actgattttt cctcgagaag accttggcat gattttgaaa atggattctt
      361 tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg
      421 ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata
      481 gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg
      541 tatcgattga ctcggcagga tccggtgacg gtgctggttt aaagttcaat aaagtcagag
      601 ttgtgagagc actggatgct gttgctctcc cacagccatt gggcgttcca aatgaaagcc
      661 aagccctaag ccagagattc actttttcac caggtcaaga catacagttg attccaccac
      721 tgatcaacct gttaatgagc attgaaccag atgtgatcta tgcaggacat gacaacacaa
      781 aacctgacac ctccagttct ttgctgacaa gtcttaatca actaggcgag aggcaacttc
      841 tttcagtagt caagtggtct aaatcattgc caggttttcg aaacttacat attgatgacc
      901 agataactct cattcagtat tcttggatga gcttaatggt gtttggtcta ggatggagat
      961 cctacaaaca tgtcagtggg cagatgctgt attttgcacc tgatctaata ctaaatgaac
     1021 agcggatgaa agaatcatca ttctattcat tatgccttac catgtggcag atcccacagg
     1081 agtttgtcaa gcttcaagtt agccaagaag agttcctctg tatgaaagta ttgttacttc
     1141 ttaatacaat tcctttggaa gggctacgaa gtcaaaccca gtttgaggag atgaggtcaa
     1201 gctacattag agagctcatc aaggcaattg gtttgaggca aaaaggagtt gtgtcgagct
     1261 cacagcgttt ctatcaactt acaaaacttc ttgataactc gcatgatctt gtcaaacagc
     1321 ttcatctgta ctgcttgaat acatttatcc agtcccgggc actgagtgtt gaatttccag
     1381 aaatgatgtc tgaagttatt gctgcacaat tacccaagat attggcaggg atggtgaaac
     1441 cccttctctt tcataaaaag gcggccgcag gtgacggtgc tggtttaatt aacatgacgg
     1501 tcgaccatga tttcaatagc gaagatattt tattccccat agaaagcatg agtagtatac
     1561 aatacgtgga gaataataac ccaaataata ttaacaacga tgttatcccg tattctctag
     1621 atatcaaaaa cactgtctta gatagtgcgg atctcaatga cattcaaaat caagaaactt
     1681 cactgaattt ggggcttcct ccactatctt tcgactctcc actgcccgta acggaaacga
     1741 taccatccac taccgataac agcttgcatt tgaaagctga tagcaacaaa aatcgcgatg
     1801 caagaactat tgaaaatgat agtgaaatta agagtactaa taatgctagt ggctctgggg
     1861 caaatcaata cacaactctt acttcacctt atcctatgaa cgacattttg tacaacatga
     1921 acaatccgtt acaatcaccg tcaccttcat cggtacctca aaatccgact ataaatcctc
     1981 ccataaatac agcaagtaac gaaactaatt tatcgcctca aacttcaaat ggtaatgaaa
     2041 ctcttatatc tcctcgagcc caacaacata cgtccattaa agataatcgt ctgtccttac
     2101 ctaatggtgc taattcgaat cttttcattg acactaaccc aaacaatttg aacgaaaaac
     2161 taagaaatca attgaactca gatacaaatt catattctaa ctccatttct aattcaaact
     2221 ccaattctac gggtaattta aattccagtt attttaattc actgaacata gactccatgc
     2281 tagatgatta cgtttctagt gatctcttat tgaatgatga tgatgatgac actaatttat
     2341 cacgccgaag atttagcgac gttataacaa accaagagct ctaagcaaat agctaaatta
     2401 tatacgaatt aatattatga ttaagtgttt acgtgagtgc gatattttta ttactatctt
     2461 atacagttgt atatactcta taaaatgagt tgtctattaa ttaacgcgat gaatgctttc
     2521 tgggtttacc tctccaacaa ctctagttta cttctcaata cattcaattg tatttgattt
     2581 gtcaatactt catcattaat caattctata gttttgtttt tctcgtttat ttccaaattt
     2641 aatgcatcaa ttttattatt caatttgtcg ttgattttgg ttaatgattt tatggtttga
     2701 tctctggcat tgattgtttg tgttagtttt tcattattga taattaaatt atttaagtta
     2761 gttatcaact cggtgttttc aagtttcaag ttttcaattt ctttagagtt tattagattt
     2821 gtcaaagttt ctgaattgct tgattggtcc ggcccaggta ttgttagcgg tttgaagcag
     2881 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     2941 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     3001 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     3061 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa
     3121 cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga
     3181 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     3241 tggggtttaa acatttaaat ggccggccca gcttttgttc cctttagtga gggttaattc
     3301 cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3361 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3421 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3481 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5461 aagtgccacc tgacgtcctc ggcgcgccga cgtctaagaa accattatta tcatgacatt
     5521 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5581 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5641 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5701 taactatgcg gcatcagagc agattgtact gagagtgcac cataccacct tttcaattca
     5761 tcattttttt tttattcttt tttttgattt cggtttcctt gaaatttttt tgattcggta
     5821 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     5881 atattgggtc agcagtacag aaccgtcgac gaagttgtaa gtggtggatc agatatcatc
     5941 attgttggca gaggactttt cgccaagggt agagatccta atactaaact cacaaattag
     6001 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     6061 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     6121 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6181 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6241 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6301 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6361 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6421 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6481 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6541 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6601 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6661 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     6721 ggggccagcg tggccgttta aacagaacga aggaaggagc acagacttag attggtatat
     6781 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     6841 agaacaaaaa cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     6901 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     6961 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     7021 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     7081 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     7141 gaagacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     7201 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggt ttagcttgcc
     7261 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
     7321 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
     7381 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
     7441 tacggctcct cgctgcggac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
     7501 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
     7561 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
     7621 cataaacaac catgacagtc aacactaaga cctatagtga gagagcagaa actcatgcct
     7681 caccagtagc acaacgatta tttcgattaa tggaactgaa gaaaaccaat ttatgtgcat
     7741 caattgatgt tgataccact aaggaattcc ttgaattaat tgataaattg ggtccttatg
     7801 tatgcttaat caagactcat attgatataa tcaatgattt ttcctatgaa tccactattg
     7861 aaccattatt agaactttca cgtaaacatc aatttatgat ttttgaagat agaaaatttg
     7921 ctgatattgg taataccgtg aagaaacaat atattggtgg agtttataaa attagtagtt
     7981 gggcagatat tactaatgct catggtgtca ctgggaatgg agtagttgaa ggattaaaac
     8041 agggagctaa agaaaccacc accaaccaag agccaagagg gttattgatg ttagctgaat
     8101 tatcatcagt gggatcatta gcatatggag aatattctca aaaaactgtt gaaattgcta
     8161 aatccgataa ggaatttgtt attggattta ttgcccaacg tgatatgggt ggacaagaag
     8221 aaggatttga ttggcttatt atgacacctg gagttggatt agatgataaa ggtgatggat
     8281 taggacaaca atatagaact gttgatgaag ttgttagcac tggaactgat attatcattg
     8341 ttggtagagg attgtttggt aaaggaagag atccagatat tgaaggtaaa aggtatagag
     8401 atgctggttg gaatgcttat ttgaaaaaga ctggccaatt ataatcagta ctgacaataa
     8461 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     8521 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     8581 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     8641 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga cctcgaattc
     8701 atcgagtgga ttcggctttg ggtaccgggc ccggcaacca aacccataca tcgggattcc
     8761 tataatacct tcgttggtct ccctaacatg taggtggcgg aggggagata tacaatagaa
     8821 cagataccag acaagacata atgggctaaa caagactaca ccaattacac tgcctcattg
     8881 atggtggtac ataacgaact aatactgtag ccctagactt gatagccatc atcatatcga
     8941 agtttcacta ccctttttcc atttgccatc tattgaagta ataataggcg catgcaactt
     9001 cttttctttt tttttctttt ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca
     9061 aaaaaatgat ggaagacact aaaggaaaaa tttaacgaca aagacagcac caacagatgt
     9121 cgttgttcca gagctgatga ggggtatctc gaagcacacg aaactttttc cttccttcat
     9181 tcacgcacac tactctctaa tgagcaacgg tatacggcct tccttccagt tacttgaatt
     9241 tgaaataaaa aaaagtttgc tgtcttgcta tcaagtata
//
