LOCUS       sequence-96595-e        5972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of a gal binding site 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 5972)
  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 5972)
  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: Gal4pBS4-Pleum
FEATURES             Location/Qualifiers
     source          1..5972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             197..913
                     /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(916..932)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(917..933)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(951..969)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1183..1202)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1184..1202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1212..1229)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1212..1228)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1221..1243)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1373..1828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1460..1479)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1670..1691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1959..2762)
                     /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(2763..2983)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3009..3028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3128..3150
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3189..3207)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3249..3752
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3789..3893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3894..3962,3963..4754)
                     /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(4112..4131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4925..5513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5414..5433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5667..5684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5837..5842,5843..5860,5861..5867)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5875..5891
                     /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     5880..5902
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5899..5915
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5899..5915
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5934..5954
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5936..5954
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggatcctt ggcgcgcccc ttaattaacg gaagcttcgg aagactctcc tccggtttaa
       61 accaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat cgtcttactt
      121 tctaactttt cttacctttt acatttcagc aatatatata tatatatttc aaggatatac
      181 cattctatct agaaaaatgt ctaaaggtga agaattattc actggtgttg tcccaatttt
      241 ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg
      301 tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta aattgccagt
      361 tccatggcca accttagtca ctactttagg ttatggtttg atgtgttttg ctagataccc
      421 agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga
      481 aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga
      541 aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa
      601 cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca tcatggctga
      661 caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg aagatggttc
      721 tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt
      781 accagacaac cattacttat cctatcaatc tgccttatcc aaagatccaa acgaaaagag
      841 agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg gtatggatga
      901 attgtacaaa taaatcgata ccgtcgacct cgagtcatgt aattagttat gtcacgctta
      961 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     1021 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     1081 ttcaaatttt tctttttttt ctgtacaaac gcgtgtacgc atgtaacatt atactgaaaa
     1141 ccttgcttga gaaggttttg ggacgctcgg gtacccaatt cgccctatag tgagtcgtat
     1201 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1261 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1321 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc
     1381 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1441 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1501 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1561 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1621 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1681 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1741 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1801 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     1861 gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt
     1921 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     1981 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     2041 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     2101 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     2161 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     2221 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     2281 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
     2341 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
     2401 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
     2461 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
     2521 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
     2581 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
     2641 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
     2701 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
     2761 atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg
     2821 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
     2881 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aagaaaccga
     2941 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctgt ggtatggtgc
     3001 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3061 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3121 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaag
     3181 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3241 ttaggacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
     3301 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
     3361 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
     3421 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
     3481 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
     3541 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
     3601 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
     3661 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
     3721 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
     3781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4861 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5041 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5101 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5161 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5221 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5281 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5341 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5401 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5461 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5521 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5581 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5641 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5701 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5761 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tacctcactc
     5821 attaggcacc ccaggcttta cactttatgc ttccggctcc tatgttgtgt ggaattgtga
     5881 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     5941 accctcacta aagggaacaa aagctggagc tc
//
