LOCUS       sequence-96607-e        6381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter comprised of derivatives from native promoters 
            galactose, cyc and leucine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6381)
  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 6381)
  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-CU1-Pleum
FEATURES             Location/Qualifiers
     source          1..6381
                     /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    475..592
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             1047..1763
                     /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(1766..1782)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1767..1783)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1801..1819)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2033..2052)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2034..2052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2062..2079)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2062..2078)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2071..2093)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2223..2678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2310..2329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2520..2541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2809..3612)
                     /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(3613..3833)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3859..3878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3978..4000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4039..4057)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4099..4602
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4639..4743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4744..4812,4813..5604)
                     /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(4962..4981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5775..6363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6264..6283
                     /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 cttaattaat agtacggatt
      481 agaagccgcc gagcgggcga cagccctccg acggaagact ctcctccgtg cgtcctcgtc
      541 ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa
      601 agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc
      661 cacaaacctt caaattaacg aatcaaatta acaaccatag gatgataatg cgattagttt
      721 tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga
      781 agcttatttg gcgagcgttg gttggtggat caagcccacg cgtaggcaat cctcgagcag
      841 atccgccagg cgtgtatata tagcgtggat ggccaggcaa ctttagtgct gacacataca
      901 ggcagtttaa accaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat
      961 cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata tatatatttc
     1021 aaggatatac cattctatct agaaaaatgt ctaaaggtga agaattattc actggtgttg
     1081 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     1141 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
     1201 aattgccagt tccatggcca accttagtca ctactttagg ttatggtttg atgtgttttg
     1261 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     1321 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     1381 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     1441 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     1501 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
     1561 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     1621 cagtcttgtt accagacaac cattacttat cctatcaatc tgccttatcc aaagatccaa
     1681 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     1741 gtatggatga attgtacaaa taaatcgata ccgtcgacct cgagtcatgt aattagttat
     1801 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
     1861 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
     1921 ttatttatat ttcaaatttt tctttttttt ctgtacaaac gcgtgtacgc atgtaacatt
     1981 atactgaaaa ccttgcttga gaaggttttg ggacgctcgg gtacccaatt cgccctatag
     2041 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2101 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2161 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     2221 cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2281 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2341 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2401 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2461 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2521 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2581 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2641 taacgcgaat tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt
     2701 acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc
     2761 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     2821 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     2881 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     2941 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     3001 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     3061 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     3121 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     3181 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     3241 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     3301 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     3361 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     3421 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     3481 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     3541 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     3601 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
     3661 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     3721 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     3781 aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctgt
     3841 ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3901 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3961 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4021 acgcgcgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4081 taatggtttc ttaggacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat
     4141 gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt
     4201 ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag
     4261 gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca
     4321 gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt
     4381 gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag
     4441 agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa
     4501 acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt tatatattta
     4561 tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc
     4621 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4681 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4741 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4801 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4861 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4921 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4981 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5041 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5101 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5161 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5221 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5281 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5341 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5401 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5461 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5521 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5581 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5641 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5701 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5761 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5821 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5881 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5941 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6001 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6061 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6121 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6181 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6241 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6301 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6361 aaaaacgcca gcaacgcggc c
//
