LOCUS       sequence-96620-e        6009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of three gal binding sites derived from native 
            galactose promoter coupled with a core promoter derived from native 
            Leucine promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6009)
  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 6009)
  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: Gal4pBS134-Pleum
FEATURES             Location/Qualifiers
     source          1..6009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             234..950
                     /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(953..969)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(954..970)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(988..1006)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1220..1239)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1221..1239)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1249..1266)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1249..1265)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1258..1280)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1410..1865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1497..1516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1707..1728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1996..2799)
                     /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(2800..3020)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3046..3065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3165..3187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3226..3244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3286..3789
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3826..3930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3931..3999,4000..4791)
                     /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(4149..4168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4962..5550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5451..5470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5704..5721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5874..5879,5880..5897,5898..5904)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5912..5928
                     /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     5917..5939
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5936..5952
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5936..5952
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5971..5991
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5973..5991
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggatcctt ggcgcgcccc ttaattaacg gaagcttcgg attagaagcc gccgagcggg
       61 cgacagccct ccgacggaag actctcctcc ggtttaaacc aatattattt aaggacctat
      121 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
      181 tttcagcaat atatatatat atatttcaag gatataccat tctatctaga aaaatgtcta
      241 aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta
      301 atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga
      361 ccttaaaatt tatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta
      421 ctttaggtta tggtttgatg tgttttgcta gatacccaga tcatatgaaa caacatgact
      481 ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg
      541 acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa
      601 tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat
      661 acaactataa ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag
      721 ttaacttcaa aattagacac aacattgaag atggttctgt tcaattagct gaccattatc
      781 aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcct
      841 atcaatctgc cttatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat
      901 ttgttactgc tgctggtatt acccatggta tggatgaatt gtacaaataa atcgataccg
      961 tcgacctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     1021 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1081 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1141 tacaaacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     1201 cgctcgggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     1261 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1321 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1381 gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc
     1441 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1501 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1561 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1621 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1681 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1741 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1801 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     1861 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg
     1921 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac
     1981 ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc
     2041 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc
     2101 aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc
     2161 caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc
     2221 ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct
     2281 cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga
     2341 caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt
     2401 caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc
     2461 tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa
     2521 ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac
     2581 tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt
     2641 gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc
     2701 acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg
     2761 atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca
     2821 ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     2881 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     2941 ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa
     3001 aaatgatgaa ttgaattgaa aagctgtggt atggtgcact ctcagtacaa tctgctctga
     3061 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3121 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3181 tcagaggttt tcaccgtcat caccgaaacg cgcgaagacg aaagggcctc gtgatacgcc
     3241 tatttttata ggttaatgtc atgataataa tggtttctta ggacggatcg cttgcctgta
     3301 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
     3361 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac
     3421 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac
     3481 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag
     3541 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat
     3601 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc
     3661 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt
     3721 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata
     3781 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3841 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3901 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3961 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4021 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4081 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4141 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4201 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4261 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4321 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4381 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4441 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4501 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4561 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4621 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4681 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4741 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4801 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4861 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4921 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4981 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5041 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5101 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5161 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5221 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5281 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5341 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5401 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5461 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5521 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5581 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5641 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5701 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5761 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5821 agtgagcgca acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac
     5881 tttatgcttc cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5941 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     6001 ctggagctc
//
