LOCUS       sequence-96608-e        5991 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 5991)
  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 5991)
  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: Gal4pBS13-Pleum
FEATURES             Location/Qualifiers
     source          1..5991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             216..932
                     /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(935..951)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(936..952)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(970..988)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1202..1221)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1203..1221)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1231..1248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1231..1247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1240..1262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1392..1847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1479..1498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1689..1710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1978..2781)
                     /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(2782..3002)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3028..3047)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3147..3169
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3208..3226)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3268..3771
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3808..3912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3913..3981,3982..4773)
                     /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(4131..4150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4944..5532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5433..5452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5686..5703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5856..5861,5862..5879,5880..5886)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5894..5910
                     /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     5899..5921
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5918..5934
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5918..5934
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5953..5973
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5955..5973
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggatcctt ggcgcgcccc ttaattaacg gaagcttcgg attagaagcc gccgagcggg
       61 cgacagccct ccggtttaaa ccaatattat ttaaggacct attgtttttt ccaataggtg
      121 gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat
      181 atatatttca aggatatacc attctatcta gaaaaatgtc taaaggtgaa gaattattca
      241 ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac aaattttctg
      301 tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa tttatttgta
      361 ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttaggt tatggtttga
      421 tgtgttttgc tagataccca gatcatatga aacaacatga ctttttcaag tctgccatgc
      481 cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca
      541 gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg
      601 attttaaaga agatggtaac attttaggtc acaaattgga atacaactat aactctcaca
      661 atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc aaaattagac
      721 acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat actccaattg
      781 gtgatggtcc agtcttgtta ccagacaacc attacttatc ctatcaatct gccttatcca
      841 aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact gctgctggta
      901 ttacccatgg tatggatgaa ttgtacaaat aaatcgatac cgtcgacctc gagtcatgta
      961 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     1021 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     1081 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaacg cgtgtacgca
     1141 tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcggg tacccaattc
     1201 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     1261 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     1321 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1381 cgaatggcgc gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1441 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1501 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1561 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1621 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1681 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1741 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1801 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat
     1861 tttctcctta cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta
     1921 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     1981 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     2041 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     2101 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     2161 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     2221 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     2281 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     2341 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     2401 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     2461 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     2521 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     2581 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     2641 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     2701 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     2761 cttatatgta gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag
     2821 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     2881 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     2941 aaaatttcaa agaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaattg
     3001 aaaagctgtg gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3061 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3121 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3181 atcaccgaaa cgcgcgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3241 tcatgataat aatggtttct taggacggat cgcttgcctg taacttacac gcgcctcgta
     3301 tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt
     3361 atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa
     3421 taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca
     3481 agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag
     3541 gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag
     3601 agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat
     3661 cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt
     3721 atatatttat attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac
     3781 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3841 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3901 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3961 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4021 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4081 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4141 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4201 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4261 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4321 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4381 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4441 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4501 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4561 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4621 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4681 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4741 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4801 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4861 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4921 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4981 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5041 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5101 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5161 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5221 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5281 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5341 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5401 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5461 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5521 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5581 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5641 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5701 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5761 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5821 aatgtgagtt acctcactca ttaggcaccc caggctttac actttatgct tccggctcct
     5881 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     5941 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct c
//
