LOCUS       sequence-224838-        8387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast mating pheromone response pathway reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8387)
  AUTHORS   Harrigan P, Madhani HD, El-Samad H
  TITLE     Real-Time Genetic Compensation Defines the Dynamic Demands of 
            Feedback Control.
  JOURNAL   Cell. 2018 Oct 18;175(3):877-886.e10. doi: 
            10.1016/j.cell.2018.09.044.
  PUBMED    30340045
REFERENCE   2  (bases 1 to 8387)
  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"; doi: 
            "10.1016/j.cell.2018.09"; journalName: "Cell"; date: "2018-10-18- 
            18"; volume: "175"; issue: "3"; pages: "877-886.e10"
COMMENT     Sequence Label: pHES943
FEATURES             Location/Qualifiers
     source          1..8387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    91..112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(127..132,133..150,151..157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    165..181
                     /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     170..192
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     189..205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     189..205
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        226..244
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        501..715
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2110..2826
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2828..2844)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2828..2844)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        3409..3752
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4571..4768
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5190..5209)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5191..5209)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5216..5233)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5216..5232)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5225..5247)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5373..5828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5460..5479)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5670..5691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6184..6203)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6303..6325
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6363..6381)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6466..6570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6571..6639,6640..7431)
                     /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(6789..6808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7602..8190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8091..8110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8344..8361
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
       61 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
      121 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
      181 atttcacaca ggaaacagct atgaccatga ttacgccaag ctcggaatta accctcacta
      241 aagggaacaa aagctgggcc ggccatttaa atgtttaaac ctactgcgcc aattgatgac
      301 aatacagacg atgataacaa accgaagtta tctgatgtag aaaaggatta aagatgctaa
      361 gagatagtga tgatatttca taaataatgt aattctatat atgttaatta ccttttttgc
      421 gaggcatatt tatggtgaag gataagtttt gaccatcaaa gaaggttaat gtggctgtgg
      481 tttcagggtc cataaagctt ttcaattcat cttttttttt tgttcttttt tttgattccg
      541 gtttctttga aatttttttg attcggtaat ctccgagcag aaggaagaac gaaggaagga
      601 gcacagactt agattggtat atatacgcat atgtggtgtt gaagaaacat gagattgccc
      661 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc
      721 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
      781 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
      841 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
      901 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
      961 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
     1021 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     1081 cggtgtggtg ggccccctcc aatattctac atgaatgaca cttccaagcg tatcatcagt
     1141 tggtgccaca ccattaatca gttttacgga gaaacaatcg ttgcatacac gtttgatgca
     1201 ggtccaaatg ctgtgttgta ctacttagct gaaaatgagt cgaaactctt tgcatttatc
     1261 tataaattgt ttggctctgt tcctggatgg gacaagaaat ttactactga gcagcttgag
     1321 gctttcaacc atcaatttga atcatctaac tttactgcac gtgaattgga tcttgagttg
     1381 caaaaggatg ttgccagagt gattttaact caagtcggtt caggcccaca agaaacaaac
     1441 gaatctttga ttgacgcaaa gactggtcta ccaaaggaat aagatcaatt cgatatgtaa
     1501 catttttctt ttcttttctt ttcctttttt tacaatagct aatttacgtt tccctacggt
     1561 attggtcgga acgaccaagc ttcaatttat aaatatctta attttaacag cagttaccac
     1621 ttgaatgaga aactttttga tgcctactct tttgcttttt catatcatga cctccctaca
     1681 tgcttcggaa tttccttatt cgtagctgtc taacagcacc acttagcaca agtacgtcca
     1741 cccgtttcat acttaaagat ctcagcagcc acccaatcta catttagggt acctgtcaca
     1801 tatcttctca aattttaggt tagctttcaa ttgtaaagtc tctttttgga tattgcacgt
     1861 ttcaagtcaa aacgcggctg taacataatt ttctgaaaca atattaaaac atagtaatga
     1921 cttaaaacaa gcgactatcg caaataagca gttcccagca agcaattaaa acgtatataa
     1981 gcgcagaata atgatctatc atcaataatt atcgtgaact tttccagtat ttacgttcat
     2041 ctttgaaaaa cggcgagcat taacaataaa agctaaaaaa accaaaaaat ttcagctcga
     2101 gaattaataa tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa
     2161 ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct
     2221 acttacggta aattgacctt aaaattgatt tgtactactg gtaaattgcc agttccatgg
     2281 ccaaccttag tcactacttt aggttatggt ttgcaatgtt ttgctagata cccagatcat
     2341 atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact
     2401 atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat
     2461 accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta
     2521 ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcactgc tgacaaacaa
     2581 aagaatggta tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg tggtgttcaa
     2641 ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac
     2701 aaccattact tatcctatca atctgcctta tccaaagatc caaacgaaaa gagagaccac
     2761 atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattgtac
     2821 aaataaggat ccactagttc tagacacctg caacatgcgt gagtaaataa gcggccgcca
     2881 ccgcggtgga gctctaagca aatagctaaa ttatatacga attaatatta tgattaagtg
     2941 tttacgtgag tgcgatattt ttattactat cttatacagt tgtatatact ctataaaatg
     3001 agttgtctat taattaacgc gatgaatgct ttctgggttt acctctccaa caactctagt
     3061 ttacttctca atacattcaa ttgtatttga tttgtcaata cttcatcatt aatcaattct
     3121 atagttttgt ttttctcgtt tatttccaaa tttaatgcat caattttatt attcaatttg
     3181 tcgttgattt tggttaatga ttttatggtt tgatctctgg cattgattgt ttgtgttagt
     3241 ttttcattat tgataattaa attatttaag ttagttatca actcggtgtt ttcaagtttc
     3301 aagttttcaa tttctttaga gtttattaga tttgtcaaag tttctgaatt gcttgattgg
     3361 tccagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     3421 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     3481 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     3541 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcgga cctgcgagca gggaaacgct
     3601 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
     3661 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
     3721 agttctcaca tcacatccga acataaacaa ccatgacagt caacactaag acctatagtg
     3781 ggagagcaga aactcatgcc tcaccagtag cacaacgatt atttcgatta atggaactga
     3841 agaaaaccaa tttatgtgca tcaattgatg ttgataccac taaggaattc cttgaattaa
     3901 ttgataaatt gggtccttat gtatgcttaa tcaagactca tattgatata atcaatgatt
     3961 tttcctatga atccactatt gaaccattat tagaactttc acgtaaacat caatttatga
     4021 tttttgaaga tagaaaattt gctgatattg gtaataccgt gaagaaacaa tatattggtg
     4081 gagtttataa aattagtagt tgggcagata ttactaatgc tcatggtgtc actgggaatg
     4141 gagtagttga aggattaaaa cagggagcta aagaaaccac caccaaccaa gagccaagag
     4201 ggttattgat gttagctgaa ttatcatcag tgggatcatt agcatatgga gaatattctc
     4261 aaaaaactgt tgaaattgct aaatccgata aggaatttgt tattggattt attgcccaac
     4321 gtgatatggg tggacaagaa gaaggatttg attggcttat tatgacacct ggagttggat
     4381 tagatgataa aggtgatgga ttaggacaac aatatagaac tgttgatgaa gttgttagca
     4441 ctggaactga tattatcatt gttggtagag gattgtttgg taaaggaaga gatccagata
     4501 ttgaaggtaa aaggtataga gatgctggtt ggaatgctta tttgaaaaag actggccaat
     4561 tataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     4621 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     4681 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
     4741 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
     4801 gtcgaaaacg agctcgaatt catcgacccg ggaatctcgg tcgtaatgat ttctataatg
     4861 acgaaaaaaa aaaaattgga aagaaaaagc ttcatggcct ttataaaaag gaactatcca
     4921 atacctcgcc agaaccaagt aacagtattt tacggggcac aaatcaagaa caataagaca
     4981 ggactgtaaa gatggacgca ttgaactcca aagaacaaca agagttccaa aaagtagtgg
     5041 aacaaaagca aatgaaggat ttcatgcgtt tgtactctaa tctggtagaa agatgtttca
     5101 cagactgtgt caatgacttc acaacatcaa agctaaccaa taaggaacaa acatgcatca
     5161 tgaagtgcgt ttaaacggcc acgctggccc cctatagtga gtcgtattac aattcactgg
     5221 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5281 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5341 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     5401 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5461 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5521 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5581 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5641 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5701 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5761 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5821 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5881 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta ttaggatctc
     5941 tacccttggc gaaaagtcct ctgccaacaa tgatgatatc tgatccacca cttacaactt
     6001 cgtcgacggt tctgtactgc tgacccaata tgcgtatata taccaatcta agtctgtgct
     6061 ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc
     6121 gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct
     6181 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6241 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6301 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     6361 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     6421 gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6481 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6541 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6601 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6661 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6721 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6781 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6841 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6901 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6961 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7021 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7081 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7141 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7201 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7261 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7321 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7381 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7441 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8221 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8281 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8341 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaa
//
