LOCUS       sequence-90264-e        6037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Induces Citrine expression from a promoter containing 3 lexA boxes 
            in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  AUTHORS   Ottoz DS, Rudolf F, Stelling J
  TITLE     Inducible, tightly regulated and growth condition-independent 
            transcription factor in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2014 Jul 17. pii: gku616.
  PUBMED    25034689
REFERENCE   2  (bases 1 to 6037)
  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"; journalName: "Nucleic 
            Acids Res. 2014 Jul 17. pii: gku616."
COMMENT     Sequence Label: FRP800_insul-(lexA-box)3-PminCYC1-Citrine-TCYC1
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        338..554
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             555..1358
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1661..2116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1798..1819)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2010..2029
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2246..2268
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2260..2277
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2261..2277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2284..2303
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2284..2302
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2669..3385
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MRSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      3426..3675
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3442..3460)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3787..3805)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3826..3842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3826..3842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3839..3861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3850..3866
                     /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)."
     promoter        complement(join(3874..3880,3881..3898,3899..3904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4057..4074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4228..4816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4987..5778,5779..5847))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5610..5629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5848..5952)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6010..6028
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgag atgctaagag atagtgatga tatttcataa ataatgtaat tctatatatg
      241 ttaattacct tttttgcgag gcatatttat ggtgaaggat aagttttgac catcaaagaa
      301 ggttaatgtg gctgtggttt cagggtccat aaagcttttc aattcatctt tttttttttt
      361 gttctttttt ttgattccgg tttctttgaa atttttttga ttcggtaatc tccgagcaga
      421 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata ggtggtgttg
      481 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acctgcagga
      541 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
      601 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
      661 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
      721 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
      781 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg
      841 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
      901 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
      961 cggcggaaga agtaacaaag gaacctagag gtcttttgat gttagcagaa ttgtcatgca
     1021 agggctccct agctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     1081 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     1141 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac
     1201 agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag
     1261 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     1321 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     1381 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccggga
     1441 atctcggtcg taatgatttc tataatgacg aaaaaaaaaa aattggaaag aaaaagcttc
     1501 atggccttta taaaaattaa ttaaggaact atccaatacc tcgccagaac caagtaacag
     1561 tattttacgg ggcacaaatc aagaacaata agacaggact gtaaagcata tgcggtgtga
     1621 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1681 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1741 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1801 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1861 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1921 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1981 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2041 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2101 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2161 tcggtgcggg cctcttcgct attacgccag ctggcgaagg ggggatgtgc tgcaaggcga
     2221 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2281 ttgtaatacg actcactata gggcgaattg gagaaacagc ggccgcaata atatataaac
     2341 ctgtataata taaccttgaa gactatattt cttttcgagc tccctaggtg ctgtatatac
     2401 tcacagcata actgtatata cacccagggt ctaggtgctg tatatactca cagcataact
     2461 gtatatacac ccagggtcta ggtgctgtat atactcacag cataactgta tatacaccca
     2521 gggtctagag catgtgctct gtatgtatat aaaactcttg ttttcttctt ttctcttctt
     2581 tccttataca ttaggacctt tgcagcataa attactatac ttctatacta gtggatcccc
     2641 cgggctgcag gaattcgata tcaagcttat gagatctaaa ggtgaagaat tattcactgg
     2701 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
     2761 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac
     2821 tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg gtttgatgtg
     2881 ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     2941 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     3001 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     3061 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     3121 ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa
     3181 cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga
     3241 tggtccagtc ttgttaccag acaaccatta cttatcctat caatctaagt tatccaaaga
     3301 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     3361 ccatggtatg gatgaattgt acaaaggatc ctaactcgag acaggcccct tttcctttgt
     3421 ctatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     3481 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     3541 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     3601 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     3661 aaggctttaa tttgcaaggt tcgcagttta cactctcatc gtcgctctca tcatcgcttc
     3721 cgttgttgtt ttccttagta gcgtctgctt ggtacccagc ttcggccgtg tacccagctt
     3781 ttgttccctt tagtgagggt taattccgag cttggcgtaa tcatggtcat agctgtttcc
     3841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg
     3901 taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc
     3961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc ggcccccctg acgagcatca
     4261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4321 gttcccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     4441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5041 atccatagtt gcctgactgc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgaaa
     5401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcttattat catgacatta
     6001 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
