LOCUS       sequence-180745-        7980 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 7980)
  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 7980)
  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: pHES822
FEATURES             Location/Qualifiers
     source          1..7980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2629..2652
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2682..3398
                     /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(3400..3416)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3400..3416)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4551..4570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4552..4570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4577..4594)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4577..4593)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4586..4608)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4734..5189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4821..4840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5031..5052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5545..5564)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5664..5686
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5724..5742)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5827..5931
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5932..6000,6001..6792)
                     /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(6150..6169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6963..7551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7452..7471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7705..7722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7839..7860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7875..7880,7881..7898,7899..7905)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7913..7929
                     /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     7918..7940
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7937..7953
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7937..7953
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(7974..7980,1..12)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cctcactaaa gggaacaaaa gctgggccgg ccatttaaat gtttaaacgg ccacgctggc
       61 catatttcta ccaatctctc aacactgagt aatggtagtt ataagaaaga gaccgagtta
      121 gggacagtta gaggcggtgg agatattcct tatggcatgt ctggcgatga taaaactttt
      181 caaacggcag ccccgatcta aaagagctga cagggaaatg gtcagaaaaa gaaacgtgca
      241 cccgcccgtc tggacgcgcc gctcacccgc acggcagaga ccaatcagta aaaatcaacg
      301 gttaacgaca ttactatata tataatatag gaagcattta atagaacagc atcgtaatat
      361 atgtgtactt tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa
      421 atagcttgtc accttacgta caatcttgat ccggagcccg tcgctttgag agaaatcaga
      481 agattccaaa agtctactga acttttgatc agaaagttgc ctttccaaag actagtcaga
      541 gaaatcgccc aagatttcaa gaccgatcta agattccaat cctctgccat cggtgctcta
      601 caggaatccg ttgaagctta cttggtctct ttgttcgaag acaccaactt ggccgctatc
      661 cacgctaagc gtgttaccat ccaaaagaag gatatcaagt tggctagaag attgagaggt
      721 gaaagatcct aagcacacag ggattgttgt acaattactt tttatgattt tagatacaca
      781 caaatctttt ctacgattac ctttcattta ttttcgtttc atgttttatt aatcattctt
      841 tttgtatatt tcacatcggg tggggttttt atattaaatt ttaataacat aatcttttaa
      901 agtttctact ttctatcctg tatattattt ccatatctat atctatattt tcaagcttgt
      961 tcacatcctt atcggaattg ttccagcgtt tttgtacaga aaatgatgtt atggtaacga
     1021 gctatatcag acctactagt cttatagagc tgtacagagt cttggaaaca taaccaacac
     1081 accaacaaga tgtcatttga ttcgttactc gacaagaatg ataagctggt aaaagtttgc
     1141 gggattcaaa ccgtcgaggc tgccgaaact gcgcttcaag caggcgctga tttgataggg
     1201 atcatatgtg tccccaacag gaagcggact atcgagagcg ctgtggctcg tgaaatatcc
     1261 aaattgattc acaaatcagg tactacaaag ctggtggggg tgttcaggaa tcaatctgtt
     1321 gaggacgtac atcggctttc tgaggaatat gaccttgaca taatccaatt acatggtgat
     1381 gaatcatggc cagagtacta taacgtcatt aagaaaccaa taatcaaaag agtcatattc
     1441 cctagagatg tcgatgttgt aacacaagtg tgtcaaagaa aacccttggt atgtctacca
     1501 ttgttcgact ccgaggcagg cggtacaggt gaaaagctgg actggtccag tatttcctcc
     1561 tgggcaagcg aacagaataa cgtattttat atactagcgg gtggactaac tgcagaaaac
     1621 gtcatggagg cggtggggtt gcctggtgta attggtgttg acgtctcagg tggtgtagag
     1681 actgacggcg tcaaagacaa tgacaagata attaaatatg tacaaaatgc aaagaaacaa
     1741 tgaattacga agagaagtct caatgaggtt ttacaaacaa tcctataata catatatgcc
     1801 acttagatat actttgaata ctgaatatac tgactcatta tattctttaa ttgtagtttc
     1861 tcaaaaatat tcatacaggt tagctatgaa ttcatgctat aatttccatt ttgttcatgt
     1921 aagatacaca tagggataaa atgtgataac taatcagcgg taccgggccc ttatattgaa
     1981 ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac
     2041 atggcattac caccatatac atatccatat acatatccat atctaatctt acttatatgt
     2101 tgtggaaatg taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc
     2161 agtaatacgc ttaactgctc attgctatat tgaagtgcgg ccgcgtgggc gtgcgtgggc
     2221 gggcgtgggc gtgcgtgggc gggcgtgggc gtgcgtgggc gtctagaccg tgcgtcctcg
     2281 tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctaat gccgcactgc tccgaacaat
     2341 aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc
     2401 cccacaaacc ttcaaattaa cgaatcaaat taacaaccat aggatgataa tgcgattagt
     2461 tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaaca
     2521 gatatataaa tggaaaagct gcataaccac tttaactaat actttcaaca ttttcagttt
     2581 gtattacttc ttattcaaat gtcataaaag tatcaacaaa aaattgttaa tatacctcta
     2641 tactttaacg tcaaggagaa aaaactatac tcgagaataa aatgtctaaa ggtgaagaat
     2701 tattcactgg tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat
     2761 tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattga
     2821 tttgtactac tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg
     2881 gtttgcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg
     2941 ccatgccaga aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca
     3001 agaccagagc tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag
     3061 gtattgattt taaagaagat ggtaacattt taggtcacaa attggaatac aactataact
     3121 ctcacaatgt ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa
     3181 ttagacacaa cattgaagat ggtggtgttc aattagctga ccattatcaa caaaatactc
     3241 caattggtga tggtccagtc ttgttaccag acaaccatta cttatcctat caatctgcct
     3301 tatccaaaga tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg
     3361 ctggtattac ccatggtatg gatgaattgt acaaataagg atccactagt tctagacacc
     3421 tgcaacatgc gtgagtaaat aagcggccgc caccgcggtg gagctctaag caaatagcta
     3481 aattatatac gaattaatat tatgattaag tgtttacgtg agtgcgatat ttttattact
     3541 atcttataca gttgtatata ctctataaaa tgagttgtct attaattaac gcgatgaatg
     3601 ctttctgggt ttacctctcc aacaactcta gtttacttct caatacattc aattgtattt
     3661 gatttgtcaa tacttcatca ttaatcaatt ctatagtttt gtttttctcg tttatttcca
     3721 aatttaatgc atcaatttta ttattcaatt tgtcgttgat tttggttaat gattttatgg
     3781 tttgatctct ggcattgatt gtttgtgtta gtttttcatt attgataatt aaattattta
     3841 agttagttat caactcggtg ttttcaagtt tcaagttttc aatttcttta gagtttatta
     3901 gatttgtcaa agtttctgaa ttgcttgatt ggtcctgtag aagagtattt gttgttgtgg
     3961 ataattgatt caatttttga gacaattgct ggaaggcgtt gaaatatcta gcatcaatct
     4021 catggttttt ttcccgagag tctcgtagat tcaattgttt taatatatct tgggaccact
     4081 cttgatttga actcatggaa attaaactgg gtgttgtgtt gtggtgtaat gattgtaccc
     4141 cctttgctta taattgtgtg gttccgatgc tgacttactg ggtattatat gtgtgcccaa
     4201 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata
     4261 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga
     4321 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga
     4381 gtcgtggcga gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc
     4441 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt
     4501 gtttgattca gaaggccggc catttaaatg tttaaacggc cacgctggcc ccctatagtg
     4561 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4621 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4681 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     4741 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4801 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4861 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4921 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4981 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5041 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5101 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5161 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     5221 tgcggtattt cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg
     5281 tgagtttagt attaggatct ctacccttgg cgaaaagtcc tctgccaaca atgatgatat
     5341 ctgatccacc acttacaact tcgtcgacgg ttctgtactg ctgacccaat atgcgtatat
     5401 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     5461 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     5521 aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5581 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5641 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5701 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5761 gataataatg gtttcttaga cgtcggcgcg ccgaggacgt caggtggcac ttttcgggga
     5821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5941 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6001 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6061 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6121 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6181 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6241 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6301 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6361 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6421 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6481 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6541 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6601 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6661 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6721 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6781 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6841 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6901 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6961 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7021 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7081 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7141 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7201 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7261 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7321 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7381 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7441 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7501 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7561 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7621 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7681 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7741 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7801 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7861 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7921 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cggaattaac
//
