LOCUS       sequence-172831-        7950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP under control of GAL1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7950)
  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 7950)
  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: pHES820
FEATURES             Location/Qualifiers
     source          1..7950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2162..2603
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2162..2279
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2574..2597
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2621..3337
                     /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(3339..3355)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3339..3355)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4497..4516)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4498..4516)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4523..4540)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4523..4539)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4532..4554)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4680..5135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4767..4786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4977..4998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5491..5510)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5610..5632
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5670..5688)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5773..5877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5878..5946,5947..6738)
                     /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(6096..6115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6909..7497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7398..7417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7651..7668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7785..7806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7821..7826,7827..7844,7845..7851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7859..7875
                     /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     7864..7886
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7883..7899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7883..7899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7920..7938
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccata tttctaccaa tctctcaaca
       61 ctgagtaatg gtagttataa gaaagagacc gagttaggga cagttagagg cggtggagat
      121 attccttatg gcatgtctgg cgatgataaa acttttcaaa cggcagcccc gatctaaaag
      181 agctgacagg gaaatggtca gaaaaagaaa cgtgcacccg cccgtctgga cgcgccgctc
      241 acccgcacgg cagagaccaa tcagtaaaaa tcaacggtta acgacattac tatatatata
      301 atataggaag catttaatag aacagcatcg taatatatgt gtactttgca gttatgacgc
      361 cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct tacgtacaat
      421 cttgatccgg agcccgtcgc tttgagagaa atcagaagat tccaaaagtc tactgaactt
      481 ttgatcagaa agttgccttt ccaaagacta gtcagagaaa tcgcccaaga tttcaagacc
      541 gatctaagat tccaatcctc tgccatcggt gctctacagg aatccgttga agcttacttg
      601 gtctctttgt tcgaagacac caacttggcc gctatccacg ctaagcgtgt taccatccaa
      661 aagaaggata tcaagttggc tagaagattg agaggtgaaa gatcctaagc acacagggat
      721 tgttgtacaa ttacttttta tgattttaga tacacacaaa tcttttctac gattaccttt
      781 catttatttt cgtttcatgt tttattaatc attctttttg tatatttcac atcgggtggg
      841 gtttttatat taaattttaa taacataatc ttttaaagtt tctactttct atcctgtata
      901 ttatttccat atctatatct atattttcaa gcttgttcac atccttatcg gaattgttcc
      961 agcgtttttg tacagaaaat gatgttatgg taacgagcta tatcagacct actagtctta
     1021 tagagctgta cagagtcttg gaaacataac caacacacca acaagatgtc atttgattcg
     1081 ttactcgaca agaatgataa gctggtaaaa gtttgcggga ttcaaaccgt cgaggctgcc
     1141 gaaactgcgc ttcaagcagg cgctgatttg atagggatca tatgtgtccc caacaggaag
     1201 cggactatcg agagcgctgt ggctcgtgaa atatccaaat tgattcacaa atcaggtact
     1261 acaaagctgg tgggggtgtt caggaatcaa tctgttgagg acgtacatcg gctttctgag
     1321 gaatatgacc ttgacataat ccaattacat ggtgatgaat catggccaga gtactataac
     1381 gtcattaaga aaccaataat caaaagagtc atattcccta gagatgtcga tgttgtaaca
     1441 caagtgtgtc aaagaaaacc cttggtatgt ctaccattgt tcgactccga ggcaggcggt
     1501 acaggtgaaa agctggactg gtccagtatt tcctcctggg caagcgaaca gaataacgta
     1561 ttttatatac tagcgggtgg actaactgca gaaaacgtca tggaggcggt ggggttgcct
     1621 ggtgtaattg gtgttgacgt ctcaggtggt gtagagactg acggcgtcaa agacaatgac
     1681 aagataatta aatatgtaca aaatgcaaag aaacaatgaa ttacgaagag aagtctcaat
     1741 gaggttttac aaacaatcct ataatacata tatgccactt agatatactt tgaatactga
     1801 atatactgac tcattatatt ctttaattgt agtttctcaa aaatattcat acaggttagc
     1861 tatgaattca tgctataatt tccattttgt tcatgtaaga tacacatagg gataaaatgt
     1921 gataactaat cagcggtacc gggcccttat attgaatttt caaaaattct tacttttttt
     1981 ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc atatacatat
     2041 ccatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat cttagcctaa
     2101 aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct atattgaagt
     2161 acggattaga agccgccgag cgggcgacag ccctccgacg gaagactctc ctccgtgcgt
     2221 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
     2281 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
     2341 ctggccccac aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga
     2401 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
     2461 taacagatat ataaatggaa aagctgcata accactttaa ctaatacttt caacattttc
     2521 agtttgtatt acttcttatt caaatgtcat aaaagtatca acaaaaaatt gttaatatac
     2581 ctctatactt taacgtcaag gagaaaaaac tatactcgag atgtctaaag gtgaagaatt
     2641 attcactggt gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt
     2701 ttctgtctcc ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaattgat
     2761 ttgtactact ggtaaattgc cagttccatg gccaacctta gtcactactt taggttatgg
     2821 tttgcaatgt tttgctagat acccagatca tatgaaacaa catgactttt tcaagtctgc
     2881 catgccagaa ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa
     2941 gaccagagct gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg
     3001 tattgatttt aaagaagatg gtaacatttt aggtcacaaa ttggaataca actataactc
     3061 tcacaatgtt tacatcactg ctgacaaaca aaagaatggt atcaaagcta acttcaaaat
     3121 tagacacaac attgaagatg gtggtgttca attagctgac cattatcaac aaaatactcc
     3181 aattggtgat ggtccagtct tgttaccaga caaccattac ttatcctatc aatctgcctt
     3241 atccaaagat ccaaacgaaa agagagacca catggtcttg ttagaatttg ttactgctgc
     3301 tggtattacc catggtatgg atgaattgta caaataagga tccactagtt ctagacacct
     3361 gcaacatgcg tgagtaaata agcggccgcc accgcggtgg agctctaagc aaatagctaa
     3421 attatatacg aattaatatt atgattaagt gtttacgtga gtgcgatatt tttattacta
     3481 tcttatacag ttgtatatac tctataaaat gagttgtcta ttaattaacg cgatgaatgc
     3541 tttctgggtt tacctctcca acaactctag tttacttctc aatacattca attgtatttg
     3601 atttgtcaat acttcatcat taatcaattc tatagttttg tttttctcgt ttatttccaa
     3661 atttaatgca tcaattttat tattcaattt gtcgttgatt ttggttaatg attttatggt
     3721 ttgatctctg gcattgattg tttgtgttag tttttcatta ttgataatta aattatttaa
     3781 gttagttatc aactcggtgt tttcaagttt caagttttca atttctttag agtttattag
     3841 atttgtcaaa gtttctgaat tgcttgattg gtcctgtaga agagtatttg ttgttgtgga
     3901 taattgattc aatttttgag acaattgctg gaaggcgttg aaatatctag catcaatctc
     3961 atggtttttt tcccgagagt ctcgtagatt caattgtttt aatatatctt gggaccactc
     4021 ttgatttgaa ctcatggaaa ttaaactggg tgttgtgttg tggtgtaatg attgtacccc
     4081 ctttgcttat aattgtgtgg ttccgatgct gacttactgg gtattatatg tgtgcccaat
     4141 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
     4201 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
     4261 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
     4321 tcgtggcgag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
     4381 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
     4441 tttgattcag aaggccggcc atttaaatgt ttaaacggcc acgctggccc aattcgccct
     4501 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4561 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4621 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     4681 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4741 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4801 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4861 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4921 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4981 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5041 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5101 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     5161 catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct
     5221 aatttgtgag tttagtatta ggatctctac ccttggcgaa aagtcctctg ccaacaatga
     5281 tgatatctga tccaccactt acaacttcgt cgacggttct gtactgctga cccaatatgc
     5341 gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta
     5401 ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg
     5461 aattgaaaag gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5521 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5581 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5641 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5701 tgtcatgata ataatggttt cttagacgtc ggcgcgccga ggacgtcagg tggcactttt
     5761 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5821 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5881 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5941 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6001 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6061 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7741 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7801 actcattagg caccccaggc tttacacttt atgcttccgg ctcctatgtt gtgtggaatt
     7861 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgga
     7921 attaaccctc actaaaggga acaaaagctg
//
