LOCUS       sequence-181030-        8703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses STE4 under control of synthetic promoter containing Zif268
            binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8703)
  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 8703)
  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: pHES940
FEATURES             Location/Qualifiers
     source          1..8703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1086..1109
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(2415..2431)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2415..2431)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3750..3769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3751..3769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3776..3793)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3776..3792)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3785..3807)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3933..4388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4020..4039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4230..4251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4744..4763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4863..4885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4923..4941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5026..5130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5131..5199,5200..5991)
                     /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     complement(5349..5368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6162..6750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6651..6670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6904..6921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7038..7059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7074..7079,7080..7097,7098..7104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7112..7128
                     /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     7117..7139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7136..7152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7136..7152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7173..7191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7517..7704
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta ttcttgtaat
       61 atttggaata atagaaaacc aagactactg cctttctttt catattatag aggaagaaat
      121 acgcacgaac acgatataga ggtaaatttt ttgcaaaaag ataccaaact ataaaagaaa
      181 tagaagacta aactacaaaa aataaagaaa gtaatacata tatccaatat gactgagtta
      241 gcagagcaag tttccatgcc tgaagggcaa caaagacgtg gtggttacgt tccacctcac
      301 atccgtaacg gtggtggcag aagaggcgat tctgctcctc gtggtggttt caacaacggt
      361 ggcggccgtg gtggtttctt cagaaacaac ggtgaccgtg gtggattcgg ctttgggtac
      421 cgggccctta tattgaattt tcaaaaattc ttactttttt tttggatgga cgcaaagaag
      481 tttaataatc atattacatg gcattaccac catatacata tccatataca tatccatatc
      541 taatcttact tatatgttgt ggaaatgtaa agagccccat tatcttagcc taaaaaaacc
      601 ttctctttgg aactttcagt aatacgctta actgctcatt gctatattga agtgcggccg
      661 cgtgggcgtg cgtgggcggg cgtgggcgtg cgtgggcggg cgtgggcgtg cgtgggcgtc
      721 tagaccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctaatgcc
      781 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
      841 attggcagta acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg
      901 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
      961 tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt aactaatact
     1021 ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa
     1081 ttgttaatat acctctatac tttaacgtca aggagaaaaa actatactcg acaataaaat
     1141 ggcagcacat cagatggact cgataacgta ttctaataat gtcacccaac agtatataca
     1201 accacaaagt ctacaggata tctctgcagt ggaggatgaa attcaaaata aaatagaggc
     1261 cgccagacaa gagagtaaac agcttcatgc tcaaataaat aaagcaaaac acaagataca
     1321 agatgcaagc ttattccaga tggccaacaa agttacttcg ttgaccaaaa ataagatcaa
     1381 cttaaagcca aatatcgtgt tgaaaggcca taataataaa atctcagatt ttcggtggag
     1441 tcgagattca aaacgtattt tgagtgcaag tcaagatggc tttatgctta tatgggacag
     1501 tgcttcaggt ttaaaacaga acgctattcc attagattct caatgggttc tttcctgcgc
     1561 tatttcgcca tcgagtactt tggtagcaag cgcaggatta aacaataact gtaccattta
     1621 tagagtttcg aaagaaaaca gagtagcgca aaacgttgcg tcaattttca aaggacatac
     1681 ttgctatatt tctgacattg aatttacaga taacgcacat atattgacag caagtgggga
     1741 tatgacatgt gccttgtggg atataccgaa agcaaagagg gtgagagaat attctgacca
     1801 tttaggtgat gttttggcat tagctattcc tgaagagcca aactcagaaa attcttcgaa
     1861 cacattcgct agctgtggat cagacgggta tacttacata tgggatagca gatctccgtc
     1921 cgctgtacaa agcttttacg ttaacgatag tgatattaat gcacttcgtt ttttcaaaga
     1981 cgggatgtcg attgttgcag gaagtgacaa tggtgcgata aatatgtatg atttaaggtc
     2041 ggactgttct attgctactt tttctctttt tcgaggttat gaagaacgta cccctacccc
     2101 tacttatatg gcagctaaca tggagtacaa taccgcgcaa tcgccacaaa ctttaaaatc
     2161 aacaagctca agctatctag acaaccaagg cgttgtttct ttagatttta gtgcatctgg
     2221 aagattgatg tactcatgct atacagacat tggttgtgtt gtgtgggatg tattaaaagg
     2281 agagattgtt ggaaaattag aaggtcatgg tggcagagtc actggtgtgc gctcgagtcc
     2341 agatgggtta gctgtatgta caggttcatg ggactcaacc atgaaaatat ggtctccagg
     2401 ttatcaatag taaggatcca ctagttctag acacctgcaa catgcgtgag taaataagcg
     2461 gccgccaccg cggtggagct ctaagcaaat agctaaatta tatacgaatt aatattatga
     2521 ttaagtgttt acgtgagtgc gatattttta ttactatctt atacagttgt atatactcta
     2581 taaaatgagt tgtctattaa ttaacgcgat gaatgctttc tgggtttacc tctccaacaa
     2641 ctctagttta cttctcaata cattcaattg tatttgattt gtcaatactt catcattaat
     2701 caattctata gttttgtttt tctcgtttat ttccaaattt aatgcatcaa ttttattatt
     2761 caatttgtcg ttgattttgg ttaatgattt tatggtttga tctctggcat tgattgtttg
     2821 tgttagtttt tcattattga taattaaatt atttaagtta gttatcaact cggtgttttc
     2881 aagtttcaag ttttcaattt ctttagagtt tattagattt gtcaaagttt ctgaattgct
     2941 tgattggtcc tgtagaagag tatttgttgt tgtggataat tgattcaatt tttgagacaa
     3001 ttgctggaag gcgttgaaat atctagcatc aatctcatgg tttttttccc gagagtctcg
     3061 tagattcaat tgttttaata tatcttggga ccactcttga tttgaactca tggaaattaa
     3121 actgggtgtt gtgttgtggt gtaatgattg tacccccttt gcttataatt gtgtggtgac
     3181 accgattatt taaagctgca gcatacgata tatatacatg tgtatatatg tatacctatg
     3241 aatgtcagta agtatgtata cgaacagtat gatactgaag atgacaaggt aatgcatcat
     3301 tctatacgtg tcattctgaa cgaggcgcgc tttccttttt tctttttgct ttttcttttt
     3361 ttttctcttg aactccagaa aaaaaatata aaagagatgg aggaacggga aaaagttagt
     3421 tgtggtgata ggtggcaagt ggtattccgt aagaacaaca agaaaagcat ttcatattat
     3481 ggctgaactg agcgaacaag tgcaaaattt aagcatcaac gacaacaacg agaatggtta
     3541 tgttcctcct cacttaagag gaaaaccaag aagtgccaga aataacagta gcaactacaa
     3601 taacaacaac ggcggctaca acggtggccg tggcggtggc agcttcttta gcaacaaccg
     3661 tcgtggtggt tacggcaacg gtggtttctt cggtggaaac aacggtggca gcggccggcc
     3721 atttaaatgt ttaaacggcc acgctggccc cctatagtga gtcgtattac aattcactgg
     3781 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3841 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3901 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     3961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4021 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4081 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4141 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4201 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4261 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4321 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4381 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4441 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta ttaggatctc
     4501 tacccttggc gaaaagtcct ctgccaacaa tgatgatatc tgatccacca cttacaactt
     4561 cgtcgacggt tctgtactgc tgacccaata tgcgtatata taccaatcta agtctgtgct
     4621 ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc
     4681 gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct
     4741 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4801 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4861 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4921 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4981 gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5041 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5101 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5161 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5221 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5281 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5341 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5401 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5461 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5521 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5581 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5641 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5701 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5761 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5821 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5881 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5941 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6001 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6301 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6781 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6841 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6901 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6961 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7021 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7081 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7141 aacagctatg accatgatta cgccaagctc ggaattaacc ctcactaaag ggaacaaaag
     7201 ctgggccggc catttaaatg tttaaacggc cacgctggcc gctgcacggt cctgttccct
     7261 agcatgtacg tgagcgtatt tccttttaaa ccacgacgct ttgtcttcat tcaacgtttc
     7321 ccattgtttt tttctactat tgctttgctg tgggaaaaac ttatcgaaag atgacgactt
     7381 tttcttaatt ctcgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
     7441 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
     7501 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
     7561 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
     7621 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
     7681 caggcaagat aaacgaaggc aaagaggctc tatatgtata taaagctgcg gatgccttct
     7741 gatttaatct agttctagtt tacttcaggt ttacttaaag ttgagataat tgaagcaaag
     7801 agaagcatcg agcaaaggat ggcgtttgtt aagagggtta cgcaggagac gaatatacag
     7861 ctggcgctgg atcttgacgg tgggtctgtt tctgtacggg agagcatact gggcaaggaa
     7921 tatgctagtg gtgatgggca gaccatccat gtgcacactg gagttgggtt tttggaccac
     7981 atgttgactg cgctggcgaa gcatggcggg tggtctctga tcctggagtg tataggggac
     8041 ttgcacatcg atgaccacca cactgttgag gactgtggga ttgcgctggg ccaagcgttc
     8101 aaggaggcgc ttggctccgt gcgtggtatc aagaggttcg ggcatgggtt tgcaccactg
     8161 gacgaggcgc tgagccgcgc tgtggttgac ttctccaata ggcctttcgc cgtggtggag
     8221 ctgggcctga agcgagaacg cataggccag ctatccacag agatgatccc gcacttcttg
     8281 gagagtttcg ccactgaggc gcgtatcacc atgcatgtgg actgtctgcg gggcaccaac
     8341 gaccaccacc gctccgaatc agctttcaag gcgctcgcca tcgccatcag agaggcaaga
     8401 acacctacgg gtcgcgatga cgttccatcc actaagggtg tcctagcata gaacacagcc
     8461 cacagctacc accatcaaca atatttatat agataacgta cacatagaaa tcacacaaac
     8521 agagtattta ttcttaacta catgaactac catcagaccg tctgggccac tatataatgt
     8581 gctattcata aacgtgatca ctctacgtag caggcgaccc caggtgaaaa tttttcagcg
     8641 agctgccaga ttgtcaggtg aaaaactgaa aaaaacttct gggcgatgag cttgtggcgg
     8701 gaa
//
