LOCUS       sequence-474139-        6298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains full length native galactose promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6298)
  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 6298)
  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: PGAL
FEATURES             Location/Qualifiers
     source          1..6298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    272..293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(308..313,314..331,332..338)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    346..362
                     /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     351..373
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     370..386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     370..386
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     405..425
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        407..425
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        497..938
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    497..614
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     909..932
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             962..1678
                     /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(1681..1697)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1682..1698)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1700..1947
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1716..1734)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1965..1984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1966..1984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1994..2011)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1994..2010)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2003..2025)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2151..2606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2238..2257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2448..2469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2737..3540)
                     /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(3541..3756)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3780..3799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3899..3921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3959..3977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4018..4521
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4558..4662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4663..4731,4732..5523)
                     /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(4881..4900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5694..6282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6183..6202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
       61 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
      121 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
      181 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      241 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      301 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      361 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
      421 aaagggaaca aaagctggag ctccgggatc cttggcgcgc cccttaatta acggaagctt
      481 cctgtttaaa ctagtacgga ttagaagccg ccgagcgggc gacagccctc cgacggaaga
      541 ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc
      601 gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga
      661 aaaattggca gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat taacaaccat
      721 aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga
      781 tgatttttga tctattaaca gatatataaa tggaaaagct gcataaccac tttaactaat
      841 actttcaaca ttttcagttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa
      901 aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaaccccgg attctagaaa
      961 aatgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg aattagatgg
     1021 tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg
     1081 taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat ggccaacctt
     1141 agtcactact ttaggttatg gtttgatgtg ttttgctaga tacccagatc atatgaaaca
     1201 acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa ctattttttt
     1261 caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg ataccttagt
     1321 taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt taggtcacaa
     1381 attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg
     1441 tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc aattagctga
     1501 ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag acaaccatta
     1561 cttatcctat caatctgcct tatccaaaga tccaaacgaa aagagagacc acatggtctt
     1621 gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt acaaataaat
     1681 cgataccgtc gacctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1741 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1801 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1861 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1921 ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt
     1981 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2041 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2101 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     2161 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2221 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2281 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2341 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2401 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2461 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2521 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2581 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     2641 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
     2701 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
     2761 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
     2821 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
     2881 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
     2941 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
     3001 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
     3061 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
     3121 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
     3181 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
     3241 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
     3301 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
     3361 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
     3421 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
     3481 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
     3541 gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca
     3601 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
     3661 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa
     3721 tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg cactctcagt
     3781 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3841 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3901 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3961 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacggat
     4021 cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat ttgggaattt
     4081 actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa ataaagaagg
     4141 tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa
     4201 agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga tatacattcg
     4261 attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc tacacagaca
     4321 agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa aggtagtatt
     4381 tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat tttttcttta
     4441 atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat ttaaattata
     4501 attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc
     4561 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4621 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4681 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4741 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4801 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4861 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4921 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4981 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5041 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5101 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5161 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5221 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5281 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5341 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5401 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5461 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5521 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5581 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5641 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5701 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5761 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5821 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5881 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5941 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6001 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6061 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6121 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6181 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6241 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcgg
//
