LOCUS       sequence-96590-e        5984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of two gal binding sites 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 5984)
  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 5984)
  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: Gal4pBS12-Pleum
FEATURES             Location/Qualifiers
     source          1..5984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             209..925
                     /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(928..944)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(929..945)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(963..981)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1195..1214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1196..1214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1224..1241)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1224..1240)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1233..1255)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1385..1840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1472..1491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1682..1703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1971..2774)
                     /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(2775..2995)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3021..3040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3140..3162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3201..3219)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3261..3764
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3801..3905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3906..3974,3975..4766)
                     /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(4124..4143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4937..5525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5426..5445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5679..5696
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5849..5854,5855..5872,5873..5879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5887..5903
                     /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     5892..5914
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5911..5927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5911..5927
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5946..5966
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5948..5966
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggatcctt ggcgcgcccc ttaattaacg gaagcttcgg attagaagcc gccgagggcg
       61 acagccgttt aaaccaatat tatttaagga cctattgttt tttccaatag gtggttagca
      121 atcgtcttac tttctaactt ttcttacctt ttacatttca gcaatatata tatatatatt
      181 tcaaggatat accattctat ctagaaaaat gtctaaaggt gaagaattat tcactggtgt
      241 tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg
      301 tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt gtactactgg
      361 taaattgcca gttccatggc caaccttagt cactacttta ggttatggtt tgatgtgttt
      421 tgctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg
      481 ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga
      541 agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa
      601 agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta
      661 catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta gacacaacat
      721 tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg
      781 tccagtcttg ttaccagaca accattactt atcctatcaa tctgccttat ccaaagatcc
      841 aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg gtattaccca
      901 tggtatggat gaattgtaca aataaatcga taccgtcgac ctcgagtcat gtaattagtt
      961 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     1021 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     1081 cgttatttat atttcaaatt tttctttttt ttctgtacaa acgcgtgtac gcatgtaaca
     1141 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gggtacccaa ttcgccctat
     1201 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1261 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1321 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1381 cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1801 tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc
     1861 ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa
     1921 gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc
     1981 tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc
     2041 ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta
     2101 gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa
     2161 cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt
     2221 tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta
     2281 gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct
     2341 ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc
     2401 accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag
     2461 tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa
     2521 ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc
     2581 aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc
     2641 agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc
     2701 atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat
     2761 gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt
     2821 aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta
     2881 agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt
     2941 caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct
     3001 gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3061 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3121 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3181 aaacgcgcga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     3241 aataatggtt tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta
     3301 atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga
     3361 ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa
     3421 aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag
     3481 cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc
     3541 gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga
     3601 agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga
     3661 aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt
     3721 tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg
     3781 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3841 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3901 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3961 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4021 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4081 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4141 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4201 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4261 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4321 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4381 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4441 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4501 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4561 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4621 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4681 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4741 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4801 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4861 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4921 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5161 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5221 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5281 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5341 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5401 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5461 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5521 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5581 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5641 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5701 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5761 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5821 gttacctcac tcattaggca ccccaggctt tacactttat gcttccggct cctatgttgt
     5881 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5941 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctc
//
