LOCUS       sequence-180946-        8715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP 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 8715)
  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 8715)
  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: pHES841
FEATURES             Location/Qualifiers
     source          1..8715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        267..674
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     3158..3181
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3211..3927
                     /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(3929..3945)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3929..3945)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5253..5272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5254..5272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5279..5296)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5279..5295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5288..5310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5436..5891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5523..5542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5733..5754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6247..6266)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6366..6388
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6426..6444)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6529..6633
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6634..6702,6703..7494)
                     /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(6852..6871)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7665..8253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8154..8173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8407..8424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8541..8562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8577..8582,8583..8600,8601..8607)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8615..8631
                     /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     8620..8642
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8639..8655
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8639..8655
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8676..8694
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tttaaatgtt taaacggcca cgctggccga ggagaacttc tagtatatct acatacctaa
       61 tattattgcc ttattaaaaa tggaatccca acgattacat caaaatccac attctcttca
      121 aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata tttataggat
      181 aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct aaggcgcctg
      241 attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg taagatgcaa
      301 gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg
      361 cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc
      421 tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg
      481 aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact
      541 tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt
      601 cttactttct aacttttctt accttttaca tttcagcaat atatatatat atatttcaag
      661 gatataccat tctaatgtct gcccctaagt taccaataga tagcggttta atataaggtt
      721 tcaacaagaa atattttgtg aaaatcaaaa gttaatagtt tcactgtcga aaatatgaaa
      781 gtttaagttt tcagttcatc ccattgtagt tatcaaattt ctgtgtttcc cgtatatgtg
      841 tgctctgtat tcatcgctat cttgaaactg gttatgatcg ttgcgcatgt gacctgtatc
      901 gctcaaaaga gcccggcacc ggtaccgggc aaaagcccct agcaagctaa aatgattcaa
      961 ctttcgtagt aatttgttca accggtcaca gtaaagtcaa tgactaactt ttgaatcgag
     1021 gatgtatttc caattgaaag caatgtatat ataaaccatt gtgttgaagc ttgtcttaca
     1081 gttttcttgt atcctcccat tgttcatata gtacttaaag taccataatc cttaaagaac
     1141 aaacttgaaa gagaatacac tatacacaac taataaataa agaataatca tggctgtgac
     1201 caagacaatt gtagttctac caggtgacca tgttggtcaa gaaatcactg aagaggccat
     1261 taaagtttta aatgctattc aggaatgtcg tccagacaag gtcaatttca agtttgatca
     1321 tcatttgatc ggtggtgctg caattgatgc cactggtgtt ccattaccag acgaagcttt
     1381 ggaggcctct aagaaagctg atgccgtgct tctgggtgct gttggtggtc caaaatgggg
     1441 tactggcgct gtcagaccag aacaaggtct tttgaaaatc cgtaaggagt tgcaattgta
     1501 tgctaatcta agaccatgta attttgcatc tgattcctta ctagatctat cgccattgaa
     1561 gcctgaaatt gcaagaggta cagatttcgt cgttgttaga gaactagtgg gtggtattta
     1621 tttcggggag agaaaagaag atgaaggtga tggtgtcgcc tgggatagcg aaaagtattc
     1681 tgtgcccgaa gttcaaagaa tcactagaat ggctgctttc atggctctgc aacacaaccc
     1741 acctttacct atttggtctc tagataaagc caacgttcta gcatcatcaa gattatggag
     1801 aaaaacagtg gaagagacga ttaagaatga attcccacaa ttgactgttc aacatcaatt
     1861 gattgattcc gctgctatga tcttggttaa gaacccaact catctaaatg gtattattat
     1921 cacaaacaat atgtttggtg atattatctc cgatgaagcc tccgtgatcc caggttcgtt
     1981 aggattactt ccatctgcct ccttggcatc tcttccagat aagaatacgg catttggttt
     2041 atatgagcca tgtcatggct ctgctccaga cttaccaaag ggaaaggtta accccgtagc
     2101 aacaatccta tctgctgcta tgatgttgaa gttgtctctg gacctgttcg aagaaggtga
     2161 aattatagaa caagcagtca agaaagtgtt ggattctggt atcagaacag cggatctaaa
     2221 gggtaccaac tctactactg aggtcggtga tgcagtagcg aaagctgtca gggaactatt
     2281 agcttagact tagtagatgt acagttgcat acataagtct gaacagaaag aagtaaatat
     2341 catatctagt tcggtttaga aatcagtgac tatattggtt ttgaccagat cgcaatcctt
     2401 gcaagcattt agtaatgact atcaaaccgg tgaatcatct gataaggcta tattagcttg
     2461 tgcattcgca tgtatcggga aacgaacggt accgggccct tatattgaat tttcaaaaat
     2521 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
     2581 accatataca tatccatata catatccata tctaatctta cttatatgtt gtggaaatgt
     2641 aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca gtaatacgct
     2701 taactgctca ttgctatatt gaagtgcggc cgcgtgggcg tgcgtgggcg ggcgtgggcg
     2761 tgcgtgggcg ggcgtgggcg tgcgtgggcg tctagaccgt gcgtcctcgt cttcaccggt
     2821 cgcgttcctg aaacgcagat gtgcctaatg ccgcactgct ccgaacaata aagattctac
     2881 aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct
     2941 tcaaattaac gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt
     3001 atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat
     3061 ggaaaagctg cataaccact ttaactaata ctttcaacat tttcagtttg tattacttct
     3121 tattcaaatg tcataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt
     3181 caaggagaaa aaactatact cgagaataaa atgtctaaag gtgaagaatt attcactggt
     3241 gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc
     3301 ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaattgat ttgtactact
     3361 ggtaaattgc cagttccatg gccaacctta gtcactactt taggttatgg tttgcaatgt
     3421 tttgctagat acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa
     3481 ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct
     3541 gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt
     3601 aaagaagatg gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt
     3661 tacatcactg ctgacaaaca aaagaatggt atcaaagcta acttcaaaat tagacacaac
     3721 attgaagatg gtggtgttca attagctgac cattatcaac aaaatactcc aattggtgat
     3781 ggtccagtct tgttaccaga caaccattac ttatcctatc aatctgcctt atccaaagat
     3841 ccaaacgaaa agagagacca catggtcttg ttagaatttg ttactgctgc tggtattacc
     3901 catggtatgg atgaattgta caaataagga tccactagtt ctagacacct gcaacatgcg
     3961 tgagtaaata agcggccgcc accgcggtgg agctctaagc aaatagctaa attatatacg
     4021 aattaatatt atgattaagt gtttacgtga gtgcgatatt tttattacta tcttatacag
     4081 ttgtatatac tctataaaat gagttgtcta ttaattaacg cgatgaatgc tttctgggtt
     4141 tacctctcca acaactctag tttacttctc aatacattca attgtatttg atttgtcaat
     4201 acttcatcat taatcaattc tatagttttg tttttctcgt ttatttccaa atttaatgca
     4261 tcaattttat tattcaattt gtcgttgatt ttggttaatg attttatggt ttgatctctg
     4321 gcattgattg tttgtgttag tttttcatta ttgataatta aattatttaa gttagttatc
     4381 aactcggtgt tttcaagttt caagttttca atttctttag agtttattag atttgtcaaa
     4441 gtttctgaat tgcttgattg gtcctgtaga agagtatttg ttgttgtgga taattgattc
     4501 aatttttgag acaattgctg gaaggcgttg aaatatctag catcaatctc atggtttttt
     4561 tcccgagagt ctcgtagatt caattgtttt aatatatctt gggaccactc ttgatttgaa
     4621 ctcatggaaa ttaaactggg tgttgtgttg tggtgtaatg attgtacccc ctttgcttat
     4681 aattgtgtgg ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa aatccttgct
     4741 taaaaagatt ctcttttttt atgatatttg tacataaact ttataaatga aattcataat
     4801 agaaacgaca cgaaattaca aaatggaata tgttcatagg gtagacgaaa ctatatacgc
     4861 aatctacata catttatcaa gaaggagaaa aaggaggatg taaaggaata caggtaagca
     4921 aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac attaatattg
     4981 acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga aactatgatt
     5041 cctaatttat atattggagg attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca
     5101 ctcaatgacc tgaccatttg atggagttta agtcaatacc tccttgaacc atttcccata
     5161 atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg acgtaggccg
     5221 gccatttaaa tgtttaaacg gccacgctgg ccccctatag tgagtcgtat tacaattcac
     5281 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5341 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     5401 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     5461 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5521 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5581 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5641 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5701 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5761 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5821 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5881 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5941 atagggtaat aactgatata attaaattga agctctaatt tgtgagttta gtattaggat
     6001 ctctaccctt ggcgaaaagt cctctgccaa caatgatgat atctgatcca ccacttacaa
     6061 cttcgtcgac ggttctgtac tgctgaccca atatgcgtat atataccaat ctaagtctgt
     6121 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
     6181 accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac
     6241 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     6301 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6361 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     6421 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6481 gacgtcggcg cgccgaggac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6541 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6601 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6661 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6721 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6781 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6841 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6901 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6961 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7021 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7081 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7141 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7201 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7261 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7321 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7381 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7441 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7501 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7801 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8281 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8341 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     8401 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     8461 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     8521 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8581 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8641 ggaaacagct atgaccatga ttacgccaag ctcggaatta accctcacta aagggaacaa
     8701 aagctgggcc ggcca
//
