LOCUS       sequence-96613-e        5990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of two gal binding sites coupled wiht a core 
            promoter derived from native Leucine promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  AUTHORS   Blazeck J, Garg R, Reed B, Alper HS
  TITLE     Controlling promoter strength and regulation in Saccharomyces 
            cerevisiae using synthetic hybrid promoters.
  JOURNAL   Biotechnol Bioeng. 2012 Nov;109(11):2884-95. doi: 10.1002/bit.24552.
            Epub 2012 May 17.
  PUBMED    22565375
REFERENCE   2  (bases 1 to 5990)
  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: "10.1002/bit.24552";
            journalName: "Biotechnol Bioeng"; date: "2012-11"; volume: "109"; 
            issue: "11"; pages: "2884-95"
COMMENT     Sequence Label: Gal4pBS34-Pleum
FEATURES             Location/Qualifiers
     source          1..5990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             215..931
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(934..950)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(935..951)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(969..987)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1201..1220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1202..1220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1230..1247)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1230..1246)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1239..1261)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1391..1846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1478..1497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1688..1709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1977..2780)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2781..3001)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3027..3046)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3146..3168
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3207..3225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3267..3770
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3807..3911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3912..3980,3981..4772)
                     /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(4130..4149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4943..5531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5432..5451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5685..5702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5855..5860,5861..5878,5879..5885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5893..5909
                     /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     5898..5920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5917..5933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5917..5933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5952..5972
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5954..5972
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggatcctt ggcgcgcccc ttaattaacg gaagcttcgg gcgacagccc tccgacggaa
       61 gactctcctc cggtttaaac caatattatt taaggaccta ttgttttttc caataggtgg
      121 ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa tatatatata
      181 tatatttcaa ggatatacca ttctatctag aaaaatgtct aaaggtgaag aattattcac
      241 tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt
      301 ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac
      361 tactggtaaa ttgccagttc catggccaac cttagtcact actttaggtt atggtttgat
      421 gtgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc
      481 agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag
      541 agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga
      601 ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa
      661 tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca aaattagaca
      721 caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata ctccaattgg
      781 tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatctg ccttatccaa
      841 agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat
      901 tacccatggt atggatgaat tgtacaaata aatcgatacc gtcgacctcg agtcatgtaa
      961 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     1021 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     1081 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaacgc gtgtacgcat
     1141 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgggt acccaattcg
     1201 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     1261 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1321 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1381 gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1801 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcctg atgcggtatt
     1861 ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg atataattaa
     1921 attgaagctc taatttgtga gtttagtata catgcattta cttataatac agttttttag
     1981 ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc
     2041 tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat
     2101 cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca
     2161 tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg
     2221 tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta
     2281 cccttagtat attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa
     2341 aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct
     2401 gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc
     2461 gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt
     2521 aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa
     2581 tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact
     2641 aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt
     2701 tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc
     2761 ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt
     2821 tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc
     2881 aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa
     2941 aaatttcaaa gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga
     3001 aaagctgtgg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3061 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3121 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3181 tcaccgaaac gcgcgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3241 catgataata atggtttctt aggacggatc gcttgcctgt aacttacacg cgcctcgtat
     3301 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
     3361 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
     3421 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
     3481 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
     3541 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
     3601 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
     3661 ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta
     3721 tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc
     3781 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3901 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3961 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4021 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4081 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4141 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4201 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4261 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4321 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4381 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4441 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4741 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4801 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5581 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5641 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5701 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5761 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5821 atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta
     5881 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5941 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc
//
