LOCUS       sequence-96619-e        6599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of activating sequence derived from native 
            galactose promoter coupled with full length native galactose 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  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 6599)
  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: UASGAL-Pgal
FEATURES             Location/Qualifiers
     source          1..6599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..153
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(306..311,312..329,330..336)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    344..360
                     /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     349..371
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     368..384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     368..384
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     403..423
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        405..423
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    484..601
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        802..1243
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    802..919
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1214..1237
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1267..1983
                     /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(1986..2002)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1987..2003)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2021..2039)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2251..2270)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2252..2270)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2280..2297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2280..2296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2289..2311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2441..2896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2528..2547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2738..2759
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3027..3830)
                     /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(3831..4051)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4077..4096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4196..4218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4257..4275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4317..4820
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4857..4961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4962..5030,5031..5822)
                     /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(5180..5199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5993..6581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6482..6501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
       61 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      121 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      181 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
      301 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
      361 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      421 agggaacaaa agctggagct ccgggatcct tggcgcgccc cttaattaac ggaagcttta
      481 gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc tcctccgtgc
      541 gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc
      601 gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa attggcagta
      661 acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg atgataatgc
      721 gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga tttttgatct
      781 attaacaggt ttaaactagt acggattaga agccgccgag cgggcgacag ccctccgacg
      841 gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc
      901 ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa
      961 gaggaaaaat tggcagtaac ctggccccac aaaccttcaa attaacgaat caaattaaca
     1021 accataggat gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga
     1081 agcgatgatt tttgatctat taacagatat ataaatggaa aagctgcata accactttaa
     1141 ctaatacttt caacattttc agtttgtatt acttcttatt caaatgtcat aaaagtatca
     1201 acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac cccggattct
     1261 agaaaaatgt ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta
     1321 gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact
     1381 tacggtaaat tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca
     1441 accttagtca ctactttagg ttatggtttg atgtgttttg ctagataccc agatcatatg
     1501 aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt
     1561 tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc
     1621 ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt
     1681 cacaaattgg aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag
     1741 aatggtatca aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta
     1801 gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac
     1861 cattacttat cctatcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg
     1921 gtcttgttag aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa
     1981 taaatcgata ccgtcgacct cgagtcatgt aattagttat gtcacgctta cattcacgcc
     2041 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     2101 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     2161 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     2221 aggttttggg acgctcgggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     2281 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2341 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2401 ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca
     2461 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2521 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2581 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2641 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2701 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2761 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2821 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2881 ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2941 accgcatagg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
     3001 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     3061 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     3121 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     3181 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     3241 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     3301 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
     3361 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
     3421 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     3481 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     3541 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     3601 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     3661 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     3721 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     3781 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     3841 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     3901 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     3961 tccttctgtt cggagattac cgaatcaaaa aaatttcaaa gaaaccgaaa tcaaaaaaaa
     4021 gaataaaaaa aaaatgatga attgaattga aaagctgtgg tatggtgcac tctcagtaca
     4081 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4141 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4201 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaagac gaaagggcct
     4261 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt aggacggatc
     4321 gcttgcctgt aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta
     4381 ctctgtgttt atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt
     4441 agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa
     4501 gcgtacttta catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga
     4561 ttaacgataa gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa
     4621 gatgaaacaa ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt
     4681 gttggcgatc cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa
     4741 tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa
     4801 ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg
     4861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6121 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcc
//
