LOCUS       sequence-180367-        8145 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 8145)
  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 8145)
  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: pHES835
FEATURES             Location/Qualifiers
     source          1..8145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     759..776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    893..914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(929..934,935..952,953..959)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    967..983
                     /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     972..994
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     991..1007
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     991..1007
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1028..1046
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1372..1559
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     3644..3667
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3697..4413
                     /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(4415..4431)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4415..4431)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5750..5769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5751..5769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5776..5793)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5776..5792)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5785..5807)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5933..6388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6020..6039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6230..6251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6744..6763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6863..6885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6923..6941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7026..7130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7131..7199,7200..7991)
                     /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(7349..7368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      841 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      901 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      961 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1021 agctcggaat taaccctcac taaagggaac aaaagctggg ccggccattt aaatgtttaa
     1081 acggccacgc tggccgctgc acggtcctgt tccctagcat gtacgtgagc gtatttcctt
     1141 ttaaaccacg acgctttgtc ttcattcaac gtttcccatt gtttttttct actattgctt
     1201 tgctgtggga aaaacttatc gaaagatgac gactttttct taattctcgt tttaagagct
     1261 tggtgagcgc taggagtcac tgccaggtat cgtttgaaca cggcattagt cagggaagtc
     1321 ataacacagt cctttcccgc aattttcttt ttctattact cttggcctcc tctagtacac
     1381 tctatatttt tttatgcctc ggtaatgatt ttcatttttt tttttccacc tagcggatga
     1441 ctcttttttt ttcttagcga ttggcattat cacataatga attatacatt atataaagta
     1501 atgtgatttc ttcgaagaat atactaaaaa atgagcaggc aagataaacg aaggcaaaga
     1561 ggctctatat gtatataaag ctgcggatgc cttctgattt aatctagttc tagtttactt
     1621 caggtttact taaagttgag ataattgaag caaagagaag catcgagcaa aggatggcgt
     1681 ttgttaagag ggttacgcag gagacgaata tacagctggc gctggatctt gacggtgggt
     1741 ctgtttctgt acgggagagc atactgggca aggaatatgc tagtggtgat gggcagacca
     1801 tccatgtgca cactggagtt gggtttttgg accacatgtt gactgcgctg gcgaagcatg
     1861 gcgggtggtc tctgatcctg gagtgtatag gggacttgca catcgatgac caccacactg
     1921 ttgaggactg tgggattgcg ctgggccaag cgttcaagga ggcgcttggc tccgtgcgtg
     1981 gtatcaagag gttcgggcat gggtttgcac cactggacga ggcgctgagc cgcgctgtgg
     2041 ttgacttctc caataggcct ttcgccgtgg tggagctggg cctgaagcga gaacgcatag
     2101 gccagctatc cacagagatg atcccgcact tcttggagag tttcgccact gaggcgcgta
     2161 tcaccatgca tgtggactgt ctgcggggca ccaacgacca ccaccgctcc gaatcagctt
     2221 tcaaggcgct cgccatcgcc atcagagagg caagaacacc tacgggtcgc gatgacgttc
     2281 catccactaa gggtgtccta gcatagaaca cagcccacag ctaccaccat caacaatatt
     2341 tatatagata acgtacacat agaaatcaca caaacagagt atttattctt aactacatga
     2401 actaccatca gaccgtctgg gccactatat aatgtgctat tcataaacgt gatcactcta
     2461 cgtagcaggc gaccccaggt gaaaattttt cagcgagctg ccagattgtc aggtgaaaaa
     2521 ctgaaaaaaa cttctgggcg atgagcttgt ggcgggaaat taagtatata aagcagttag
     2581 tttcttctgc ttcttgtggt tctgggtatt cttgtaatat ttggaataat agaaaaccaa
     2641 gactactgcc tttcttttca tattatagag gaagaaatac gcacgaacac gatatagagg
     2701 taaatttttt gcaaaaagat accaaactat aaaagaaata gaagactaaa ctacaaaaaa
     2761 taaagaaagt aatacatata tccaatatga ctgagttagc agagcaagtt tccatgcctg
     2821 aagggcaaca aagacgtggt ggttacgttc cacctcacat ccgtaacggt ggtggcagaa
     2881 gaggcgattc tgctcctcgt ggtggtttca acaacggtgg cggccgtggt ggtttcttca
     2941 gaaacaacgg tgaccgtggt ggattcggct ttgggtaccg ggcccttata ttgaattttc
     3001 aaaaattctt actttttttt tggatggacg caaagaagtt taataatcat attacatggc
     3061 attaccacca tatacatatc catatacata tccatatcta atcttactta tatgttgtgg
     3121 aaatgtaaag agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa
     3181 tacgcttaac tgctcattgc tatattgaag tgcggccgcg tgggcgtgcg tgggcgggcg
     3241 tgggcgtgcg tgggcgggcg tgggcgtgcg tgggcgtcta gaccgtgcgt cctcgtcttc
     3301 accggtcgcg ttcctgaaac gcagatgtgc ctaatgccgc actgctccga acaataaaga
     3361 ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac
     3421 aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga ttagtttttt
     3481 agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat
     3541 ataaatggaa aagctgcata accactttaa ctaatacttt caacattttc agtttgtatt
     3601 acttcttatt caaatgtcat aaaagtatca acaaaaaatt gttaatatac ctctatactt
     3661 taacgtcaag gagaaaaaac tatactcgag aataaaatgt ctaaaggtga agaattattc
     3721 actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct
     3781 gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa attgatttgt
     3841 actactggta aattgccagt tccatggcca accttagtca ctactttagg ttatggtttg
     3901 caatgttttg ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg
     3961 ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc
     4021 agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt
     4081 gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac
     4141 aatgtttaca tcactgctga caaacaaaag aatggtatca aagctaactt caaaattaga
     4201 cacaacattg aagatggtgg tgttcaatta gctgaccatt atcaacaaaa tactccaatt
     4261 ggtgatggtc cagtcttgtt accagacaac cattacttat cctatcaatc tgccttatcc
     4321 aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt
     4381 attacccatg gtatggatga attgtacaaa taaggatcca ctagttctag acacctgcaa
     4441 catgcgtgag taaataagcg gccgccaccg cggtggagct ctaagcaaat agctaaatta
     4501 tatacgaatt aatattatga ttaagtgttt acgtgagtgc gatattttta ttactatctt
     4561 atacagttgt atatactcta taaaatgagt tgtctattaa ttaacgcgat gaatgctttc
     4621 tgggtttacc tctccaacaa ctctagttta cttctcaata cattcaattg tatttgattt
     4681 gtcaatactt catcattaat caattctata gttttgtttt tctcgtttat ttccaaattt
     4741 aatgcatcaa ttttattatt caatttgtcg ttgattttgg ttaatgattt tatggtttga
     4801 tctctggcat tgattgtttg tgttagtttt tcattattga taattaaatt atttaagtta
     4861 gttatcaact cggtgttttc aagtttcaag ttttcaattt ctttagagtt tattagattt
     4921 gtcaaagttt ctgaattgct tgattggtcc tgtagaagag tatttgttgt tgtggataat
     4981 tgattcaatt tttgagacaa ttgctggaag gcgttgaaat atctagcatc aatctcatgg
     5041 tttttttccc gagagtctcg tagattcaat tgttttaata tatcttggga ccactcttga
     5101 tttgaactca tggaaattaa actgggtgtt gtgttgtggt gtaatgattg tacccccttt
     5161 gcttataatt gtgtggtgac accgattatt taaagctgca gcatacgata tatatacatg
     5221 tgtatatatg tatacctatg aatgtcagta agtatgtata cgaacagtat gatactgaag
     5281 atgacaaggt aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc tttccttttt
     5341 tctttttgct ttttcttttt ttttctcttg aactccagaa aaaaaatata aaagagatgg
     5401 aggaacggga aaaagttagt tgtggtgata ggtggcaagt ggtattccgt aagaacaaca
     5461 agaaaagcat ttcatattat ggctgaactg agcgaacaag tgcaaaattt aagcatcaac
     5521 gacaacaacg agaatggtta tgttcctcct cacttaagag gaaaaccaag aagtgccaga
     5581 aataacagta gcaactacaa taacaacaac ggcggctaca acggtggccg tggcggtggc
     5641 agcttcttta gcaacaaccg tcgtggtggt tacggcaacg gtggtttctt cggtggaaac
     5701 aacggtggca gcggccggcc atttaaatgt ttaaacggcc acgctggccc cctatagtga
     5761 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5821 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5881 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     5941 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6001 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6061 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6121 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6181 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6241 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6301 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6361 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     6421 gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt
     6481 gagtttagta ttaggatctc tacccttggc gaaaagtcct ctgccaacaa tgatgatatc
     6541 tgatccacca cttacaactt cgtcgacggt tctgtactgc tgacccaata tgcgtatata
     6601 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     6661 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     6721 aaggtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     6781 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6841 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     6901 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6961 ataataatgg tttcttagac gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa
     7021 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7081 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7141 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7201 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7261 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7321 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7381 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7441 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7501 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7561 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7621 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7681 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7741 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7801 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7861 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7921 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7981 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8041 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8101 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaa
//
