LOCUS       sequence-180365-        9075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses progesterone-inducible synthetic 
            transcription factor in yeast. 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 9075)
  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 9075)
  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: pHES830
FEATURES             Location/Qualifiers
     source          1..9075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    179..197
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    200..218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3243..3261)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3282..3298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3282..3298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3295..3317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3306..3322
                     /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(3330..3336,3337..3354,3355..3360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3375..3396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3513..3530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3684..4272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3764..3783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4443..5234,5235..5303))
                     /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     5066..5085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5304..5408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5493..5511
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5549..5571)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5671..5690
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6046..6501)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6183..6204)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6395..6414
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6627..6649
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6641..6658
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6642..6658
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6665..6684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6665..6683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7250..7593
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      8412..8609
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt
       61 ccttcattca cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac
      121 ttgaatttga aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgaatc
      181 cctatcagtg atagagatct ccctatcagt gatagagatt cttccttgtt tctttttctg
      241 caggctagca ataaaatggg tacccgccca tatgcttgcc ctgtcgagtc ctgcgatcgc
      301 cgcttttctc gctcggatga gcttacccgc catatccgca tccataccgg tcagaagccc
      361 ttccagtgtc gaatctgcat gcgtaacttc agtcgtagtg accaccttac cacccacatc
      421 cgcacccaca caggcgagaa gccttttgcc tgtgacattt gtgggaggaa gtttgccagg
      481 agtgatgaac gcaagaggca taccaaaatc catacaggta gatccggtga cggtgctggt
      541 ttaaagttca ataaagtcag agttgtgaga gcactggatg ctgttgctct cccacagcca
      601 ttgggcgttc caaatgaaag ccaagcccta agccagagat tcactttttc accaggtcaa
      661 gacatacagt tgattccacc actgatcaac ctgttaatga gcattgaacc agatgtgatc
      721 tatgcaggac atgacaacac aaaacctgac acctccagtt ctttgctgac aagtcttaat
      781 caactaggcg agaggcaact tctttcagta gtcaagtggt ctaaatcatt gccaggtttt
      841 cgaaacttac atattgatga ccagataact ctcattcagt attcttggat gagcttaatg
      901 gtgtttggtc taggatggag atcctacaaa catgtcagtg ggcagatgct gtattttgca
      961 cctgatctaa tactaaatga acagcggatg aaagaatcat cattctattc attatgcctt
     1021 accatgtggc agatcccaca ggagtttgtc aagcttcaag ttagccaaga agagttcctc
     1081 tgtatgaaag tattgttact tcttaataca attcctttgg aagggctacg aagtcaaacc
     1141 cagtttgagg agatgaggtc aagctacatt agagagctca tcaaggcaat tggtttgagg
     1201 caaaaaggag ttgtgtcgag ctcacagcgt ttctatcaac ttacaaaact tcttgataac
     1261 tcgcatgatc ttgtcaaaca gcttcatctg tactgcttga atacatttat ccagtcccgg
     1321 gcactgagtg ttgaatttcc agaaatgatg tctgaagtta ttgctgcaca attacccaag
     1381 atattggcag ggatggtgaa accccttctc tttcataaaa aggcggccgc aggtgacggt
     1441 gctggtttaa ttaacatgac ggtcgaccat gatttcaata gcgaagatat tttattcccc
     1501 atagaaagca tgagtagtat acaatacgtg gagaataata acccaaataa tattaacaac
     1561 gatgttatcc cgtattctct agatatcaaa aacactgtct tagatagtgc ggatctcaat
     1621 gacattcaaa atcaagaaac ttcactgaat ttggggcttc ctccactatc tttcgactct
     1681 ccactgcccg taacggaaac gataccatcc actaccgata acagcttgca tttgaaagct
     1741 gatagcaaca aaaatcgcga tgcaagaact attgaaaatg atagtgaaat taagagtact
     1801 aataatgcta gtggctctgg ggcaaatcaa tacacaactc ttacttcacc ttatcctatg
     1861 aacgacattt tgtacaacat gaacaatccg ttacaatcac cgtcaccttc atcggtacct
     1921 caaaatccga ctataaatcc tcccataaat acagcaagta acgaaactaa tttatcgcct
     1981 caaacttcaa atggtaatga aactcttata tctcctcgag cccaacaaca tacgtccatt
     2041 aaagataatc gtctgtcctt acctaatggt gctaattcga atcttttcat tgacactaac
     2101 ccaaacaatt tgaacgaaaa actaagaaat caattgaact cagatacaaa ttcatattct
     2161 aactccattt ctaattcaaa ctccaattct acgggtaatt taaattccag ttattttaat
     2221 tcactgaaca tagactccat gctagatgat tacgtttcta gtgatctctt attgaatgat
     2281 gatgatgatg acactaattt atcacgccga agatttagcg acgttataac aaaccaagag
     2341 ctctaagcaa atagctaaat tatatacgaa ttaatattat gattaagtgt ttacgtgagt
     2401 gcgatatttt tattactatc ttatacagtt gtatatactc tataaaatga gttgtctatt
     2461 aattaacgcg atgaatgctt tctgggttta cctctccaac aactctagtt tacttctcaa
     2521 tacattcaat tgtatttgat ttgtcaatac ttcatcatta atcaattcta tagttttgtt
     2581 tttctcgttt atttccaaat ttaatgcatc aattttatta ttcaatttgt cgttgatttt
     2641 ggttaatgat tttatggttt gatctctggc attgattgtt tgtgttagtt tttcattatt
     2701 gataattaaa ttatttaagt tagttatcaa ctcggtgttt tcaagtttca agttttcaat
     2761 ttctttagag tttattagat ttgtcaaagt ttctgaattg cttgattggt ccggcccagg
     2821 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     2881 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     2941 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     3001 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     3061 caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc
     3121 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     3181 tgaacgttac agaaaagcag gctggggttt aaacatttaa atggccggcc cagcttttgt
     3241 tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5401 gttccgcgca catttccccg aaaagtgcca cctgacgtcc tcggcgcgcc gacgtctaag
     5461 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5521 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5581 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5641 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5701 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
     5761 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     5821 acttagattg gtatatatac gcatattggg tcagcagtac agaaccgtcg acgaagttgt
     5881 aagtggtgga tcagatatca tcattgttgg cagaggactt ttcgccaagg gtagagatcc
     5941 taatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta ccctatgcgg
     6001 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     6061 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     6121 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     6181 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     6241 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     6301 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     6361 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     6421 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     6481 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6541 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6601 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     6661 attgtaatac gactcactat agggggccag cgtggccgtt taaacagaac gaaggaagga
     6721 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     6781 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc
     6841 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     6901 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     6961 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     7021 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     7081 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
     7141 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     7201 cggtgtggtg gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
     7261 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
     7321 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
     7381 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcgg acctgcgagc agggaaacgc
     7441 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
     7501 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
     7561 cagttctcac atcacatccg aacataaaca accatgacag tcaacactaa gacctatagt
     7621 gagagagcag aaactcatgc ctcaccagta gcacaacgat tatttcgatt aatggaactg
     7681 aagaaaacca atttatgtgc atcaattgat gttgatacca ctaaggaatt ccttgaatta
     7741 attgataaat tgggtcctta tgtatgctta atcaagactc atattgatat aatcaatgat
     7801 ttttcctatg aatccactat tgaaccatta ttagaacttt cacgtaaaca tcaatttatg
     7861 atttttgaag atagaaaatt tgctgatatt ggtaataccg tgaagaaaca atatattggt
     7921 ggagtttata aaattagtag ttgggcagat attactaatg ctcatggtgt cactgggaat
     7981 ggagtagttg aaggattaaa acagggagct aaagaaacca ccaccaacca agagccaaga
     8041 gggttattga tgttagctga attatcatca gtgggatcat tagcatatgg agaatattct
     8101 caaaaaactg ttgaaattgc taaatccgat aaggaatttg ttattggatt tattgcccaa
     8161 cgtgatatgg gtggacaaga agaaggattt gattggctta ttatgacacc tggagttgga
     8221 ttagatgata aaggtgatgg attaggacaa caatatagaa ctgttgatga agttgttagc
     8281 actggaactg atattatcat tgttggtaga ggattgtttg gtaaaggaag agatccagat
     8341 attgaaggta aaaggtatag agatgctggt tggaatgctt atttgaaaaa gactggccaa
     8401 ttataatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt
     8461 ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc
     8521 gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca
     8581 atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag
     8641 tgtcgaaaac gacctcgaat tcatcgagtg gattcggctt tgggtaccgg gcccggcaac
     8701 caaacccata catcgggatt cctataatac cttcgttggt ctccctaaca tgtaggtggc
     8761 ggaggggaga tatacaatag aacagatacc agacaagaca taatgggcta aacaagacta
     8821 caccaattac actgcctcat tgatggtggt acataacgaa ctaatactgt agccctagac
     8881 ttgatagcca tcatcatatc gaagtttcac tacccttttt ccatttgcca tctattgaag
     8941 taataatagg cgcatgcaac ttcttttctt tttttttctt ttctctctcc cccgttgttg
     9001 tctcaccata tccgcaatga caaaaaaatg atggaagaca ctaaaggaaa aatttaacga
     9061 caaagacagc accaa
//
