LOCUS       sequence-96610-e        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of activating sequence derived from native 
            galactose promoter 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 6706)
  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 6706)
  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-Pleum
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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     368..384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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"
     CDS             931..1647
                     /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(1650..1666)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1651..1667)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1685..1703)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1917..1936)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1918..1936)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1946..1963)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1946..1962)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1955..1977)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2107..2562
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2194..2213)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2404..2425
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2693..3496)
                     /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(3497..3717)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3743..3762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3862..3884
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3923..3941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3983..4486
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4523..4627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4628..4696,4697..5488)
                     /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(4846..4865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5659..6247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6148..6167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6571..6576,6577..6594,6595..6601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6609..6625
                     /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     6633..6649
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6670..6688
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
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 ttaaaccaat attatttaag gacctattgt tttttccaat aggtggttag
      841 caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata tatatatata
      901 tttcaaggat ataccattct atctagaaaa atgtctaaag gtgaagaatt attcactggt
      961 gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc
     1021 ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaatttat ttgtactact
     1081 ggtaaattgc cagttccatg gccaacctta gtcactactt taggttatgg tttgatgtgt
     1141 tttgctagat acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa
     1201 ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct
     1261 gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt
     1321 aaagaagatg gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt
     1381 tacatcatgg ctgacaaaca aaagaatggt atcaaagtta acttcaaaat tagacacaac
     1441 attgaagatg gttctgttca attagctgac cattatcaac aaaatactcc aattggtgat
     1501 ggtccagtct tgttaccaga caaccattac ttatcctatc aatctgcctt atccaaagat
     1561 ccaaacgaaa agagagacca catggtcttg ttagaatttg ttactgctgc tggtattacc
     1621 catggtatgg atgaattgta caaataaatc gataccgtcg acctcgagtc atgtaattag
     1681 ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag
     1741 ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag
     1801 aacgttattt atatttcaaa tttttctttt ttttctgtac aaacgcgtgt acgcatgtaa
     1861 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgggtaccc aattcgccct
     1921 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1981 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2041 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2101 ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2161 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2221 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2281 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2341 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2401 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2461 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2521 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc ggtattttct
     2581 ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg
     2641 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
     2701 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
     2761 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
     2821 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta
     2881 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
     2941 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
     3001 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
     3061 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
     3121 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
     3181 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa
     3241 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     3301 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     3361 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
     3421 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
     3481 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg
     3541 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
     3601 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
     3661 ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aattgaaaag
     3721 ctgtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3781 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3841 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3901 cgaaacgcgc gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3961 ataataatgg tttcttagga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
     4021 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
     4081 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
     4141 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
     4201 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
     4261 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
     4321 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     4381 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     4441 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
     4501 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4561 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4621 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4681 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4741 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4801 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4861 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4921 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4981 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5041 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5101 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5161 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5221 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5281 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5341 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5401 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5461 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5521 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5581 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5641 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5761 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6241 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6301 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6361 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6421 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6481 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6541 gagttacctc actcattagg caccccaggc tttacacttt atgcttccgg ctcctatgtt
     6601 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6661 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctc
//
