LOCUS       sequence-96617-e        6429 bp    DNA     circular SYN 15-APR-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 6429)
  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 6429)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-PCYC
FEATURES             Location/Qualifiers
     source          1..6429
                     /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    484..601
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             1095..1811
                     /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(1814..1830)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1815..1831)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1849..1867)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2081..2100)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2082..2100)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2110..2127)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2110..2126)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2119..2141)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2271..2726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2358..2377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2568..2589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2857..3660)
                     /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(3661..3881)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3907..3926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4026..4048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4087..4105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4147..4650
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4687..4791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4792..4860,4861..5652)
                     /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(5010..5029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6312..6331
                     /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 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 ttaaacattt ggcgagcgtt ggttggtgga tcaagcccac gcgtaggcaa
      841 tcctcgagca gatccgccag gcgtgtatat atagcgtgga tggccaggca actttagtgc
      901 tgacacatac aggcatatat atatgtgtgc gacgacacat gatcatatgg catgcatgtg
      961 ctctgtatgt atataaaact cttgttttct tcttttctct aaatattctt tccttataca
     1021 ttaggacctt tgcagcataa attactatac ttctatagac acgcaaacac aaatacacac
     1081 actaatctag aaaaatgtct aaaggtgaag aattattcac tggtgttgtc ccaattttgg
     1141 ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg
     1201 atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa ttgccagttc
     1261 catggccaac cttagtcact actttaggtt atggtttgat gtgttttgct agatacccag
     1321 atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat gttcaagaaa
     1381 gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc aagtttgaag
     1441 gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa gatggtaaca
     1501 ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc atggctgaca
     1561 aacaaaagaa tggtatcaaa gttaacttca aaattagaca caacattgaa gatggttctg
     1621 ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac
     1681 cagacaacca ttacttatcc tatcaatctg ccttatccaa agatccaaac gaaaagagag
     1741 accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt atggatgaat
     1801 tgtacaaata aatcgatacc gtcgacctcg agtcatgtaa ttagttatgt cacgcttaca
     1861 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     1921 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
     1981 caaatttttc ttttttttct gtacaaacgc gtgtacgcat gtaacattat actgaaaacc
     2041 ttgcttgaga aggttttggg acgctcgggt acccaattcg ccctatagtg agtcgtatta
     2101 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2161 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2221 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg
     2281 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2341 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2461 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2521 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2581 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2641 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2701 taacaaaata ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     2761 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
     2821 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
     2881 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
     2941 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
     3001 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
     3061 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
     3121 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
     3181 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
     3241 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
     3301 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
     3361 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
     3421 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
     3481 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
     3541 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
     3601 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
     3661 gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca
     3721 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
     3781 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaaa gaaaccgaaa
     3841 tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga aaagctgtgg tatggtgcac
     3901 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3961 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4021 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaagac
     4081 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4141 aggacggatc gcttgcctgt aacttacacg cgcctcgtat cttttaatga tggaataatt
     4201 tgggaattta ctctgtgttt atttattttt atgttttgta tttggatttt agaaagtaaa
     4261 taaagaaggt agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt
     4321 tcaacaaaaa gcgtacttta catatatatt tattagacaa gaaaagcaga ttaaatagat
     4381 atacattcga ttaacgataa gtaaaatgta aaatcacagg attttcgtgt gtggtcttct
     4441 acacagacaa gatgaaacaa ttcggcatta atacctgaga gcaggaagag caagataaaa
     4501 ggtagtattt gttggcgatc cccctagagt cttttacatc ttcggaaaac aaaaactatt
     4561 ttttctttaa tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt
     4621 taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga
     4681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4741 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5941 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     6001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6421 aacgcggcc
//
