LOCUS       sequence-96621-e        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a promoter of activating sequence derived from native gal 
            promoter coupled with core promoter derived from native Leucine 
            promoter. Spacer between core and activating sequence was used. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  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 6293)
  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-A9-Pleum
FEATURES             Location/Qualifiers
     source          1..6293
                     /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             959..1675
                     /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(1678..1694)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1679..1695)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1713..1731)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1945..1964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1946..1964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1974..1991)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1974..1990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1983..2005)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2135..2590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2222..2241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2432..2453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2721..3524)
                     /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(3525..3745)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3771..3790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3890..3912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3951..3969)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4011..4514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4551..4655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4656..4724,4725..5516)
                     /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(4874..4893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5687..6275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6176..6195
                     /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 agcttcatgt acttacccaa ttaggaaatt tacatggttt aaaccaatat tatttaagga
      841 cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt
      901 ttacatttca gcaatatata tatatatatt tcaaggatat accattctat ctagaaaaat
      961 gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
     1021 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
     1081 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
     1141 cactacttta ggttatggtt tgatgtgttt tgctagatac ccagatcata tgaaacaaca
     1201 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     1261 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     1321 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     1381 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
     1441 caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca
     1501 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     1561 atcctatcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     1621 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aataaatcga
     1681 taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     1741 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     1801 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     1861 ttctgtacaa acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     1921 tgggacgctc gggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     1981 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2041 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2101 caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc
     2161 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2221 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2281 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2341 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2401 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2461 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2521 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2581 tttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2641 tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     2701 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     2761 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     2821 cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt tctatactgt
     2881 tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg
     2941 taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg
     3001 tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg
     3061 catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc
     3121 tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc
     3181 cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca
     3241 gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc
     3301 ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa
     3361 attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat
     3421 gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga
     3481 ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc
     3541 ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca
     3601 acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     3661 tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa
     3721 aaaaaaaatg atgaattgaa ttgaaaagct gtggtatggt gcactctcag tacaatctgc
     3781 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3841 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3901 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga agacgaaagg gcctcgtgat
     3961 acgcctattt ttataggtta atgtcatgat aataatggtt tcttaggacg gatcgcttgc
     4021 ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt
     4081 gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga
     4141 gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac
     4201 tttacatata tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg
     4261 ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa
     4321 acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc
     4381 gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct ttaatttctt
     4441 tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt ataattattt
     4501 ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4561 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4621 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4681 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4741 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4801 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4861 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4921 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4981 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5041 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5101 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5161 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5221 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5281 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5341 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5401 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5461 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5521 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5581 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5641 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5761 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5821 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5881 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5941 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6001 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6061 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6121 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6181 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6241 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcc
//
