LOCUS       sequence-96587-e        5965 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 5965)
  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 5965)
  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: Gal4pBS2-Pleum
FEATURES             Location/Qualifiers
     source          1..5965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             190..906
                     /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(909..925)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(910..926)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(944..962)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1176..1195)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1177..1195)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1205..1222)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1205..1221)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1214..1236)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1366..1821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1453..1472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1663..1684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1952..2755)
                     /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(2756..2976)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3002..3021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3121..3143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3182..3200)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3242..3745
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3782..3886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3887..3955,3956..4747)
                     /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(4105..4124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4918..5506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5407..5426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5660..5677
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5830..5835,5836..5853,5854..5860)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5868..5884
                     /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     5873..5895
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5892..5908
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5892..5908
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5927..5947
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5929..5947
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggatcctt ggcgcgcccc ttaattaacg gaagcttggc gacagccgtt taaaccaata
       61 ttatttaagg acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact
      121 tttcttacct tttacatttc agcaatatat atatatatat ttcaaggata taccattcta
      181 tctagaaaaa tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa
      241 ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct
      301 acttacggta aattgacctt aaaatttatt tgtactactg gtaaattgcc agttccatgg
      361 ccaaccttag tcactacttt aggttatggt ttgatgtgtt ttgctagata cccagatcat
      421 atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact
      481 atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat
      541 accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta
      601 ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcatggc tgacaaacaa
      661 aagaatggta tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa
      721 ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac
      781 aaccattact tatcctatca atctgcctta tccaaagatc caaacgaaaa gagagaccac
      841 atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattgtac
      901 aaataaatcg ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac
      961 gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1021 tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat
     1081 ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac attatactga aaaccttgct
     1141 tgagaaggtt ttgggacgct cgggtaccca attcgcccta tagtgagtcg tattacgcgc
     1201 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     1261 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1321 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcgacgcg ccctgtagcg
     1381 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1441 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1501 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1561 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1621 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1681 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1741 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1801 aaatattaac gtttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1861 ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt tgtgagttta
     1921 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     1981 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     2041 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     2101 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     2161 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     2221 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     2281 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     2341 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     2401 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     2461 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     2521 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     2581 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     2641 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     2701 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     2761 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     2821 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     2881 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa
     2941 aaaaagaata aaaaaaaaat gatgaattga attgaaaagc tgtggtatgg tgcactctca
     3001 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3061 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3121 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aagacgaaag
     3181 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttaggac
     3241 ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa taatttggga
     3301 atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa gtaaataaag
     3361 aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac
     3421 aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa tagatataca
     3481 ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt cttctacaca
     3541 gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga taaaaggtag
     3601 tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa ctattttttc
     3661 tttaatttct ttttttactt tctattttta atttatatat ttatattaaa aaatttaaat
     3721 tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt
     3781 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3841 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3901 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3961 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4021 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4081 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4141 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4201 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4261 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4321 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4381 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4441 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4501 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4561 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4621 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4681 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4741 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4801 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4861 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4921 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4981 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5041 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5101 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5161 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5221 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5281 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5341 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5401 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5461 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5521 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5581 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5641 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     5701 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     5761 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttacctca ctcattaggc
     5821 accccaggct ttacacttta tgcttccggc tcctatgttg tgtggaattg tgagcggata
     5881 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     5941 ctaaagggaa caaaagctgg agctc
//
