LOCUS       sequence-90263-e        6272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Induces Citrine expression from a promoter containing 8 lexA boxes 
            in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6272)
  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 6272)
  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: FRP795_insul-(lexA-box)8-PminCYC1-Citrine-TCYC1
FEATURES             Location/Qualifiers
     source          1..6272
                     /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             2904..3620
                     /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      3661..3910
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3677..3695)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(4022..4040)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4061..4077)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4061..4077)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4074..4096)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4085..4101
                     /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(4109..4115,4116..4133,4134..4139))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4292..4309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4463..5051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5222..6013,6014..6082))
                     /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     5845..5864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6083..6187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6245..6263
                     /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 gggtctaggt gctgtatata ctcacagcat aactgtatat acacccaggg tctaggtgct
     2581 gtatatactc acagcataac tgtatataca cccagggtct aggtgctgta tatactcaca
     2641 gcataactgt atatacaccc agggtctagg tgctgtatat actcacagca taactgtata
     2701 tacacccagg gtctaggtgc tgtatatact cacagcataa ctgtatatac acccagggtc
     2761 tagagcatgt gctctgtatg tatataaaac tcttgttttc ttcttttctc ttctttcctt
     2821 atacattagg acctttgcag cataaattac tatacttcta tactagtgga tcccccgggc
     2881 tgcaggaatt cgatatcaag cttatgagat ctaaaggtga agaattattc actggtgttg
     2941 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     3001 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
     3061 aattgccagt tccatggcca accttagtca ctactttagg ttatggtttg atgtgttttg
     3121 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     3181 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     3241 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     3301 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     3361 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
     3421 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     3481 cagtcttgtt accagacaac cattacttat cctatcaatc taagttatcc aaagatccaa
     3541 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     3601 gtatggatga attgtacaaa ggatcctaac tcgagacagg ccccttttcc tttgtctata
     3661 tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg
     3721 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat
     3781 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acaaacgcgt
     3841 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     3901 tttaatttgc aaggttcgca gtttacactc tcatcgtcgc tctcatcatc gcttccgttg
     3961 ttgttttcct tagtagcgtc tgcttggtac ccagcttcgg ccgtgtaccc agcttttgtt
     4021 ccctttagtg agggttaatt ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4081 gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag
     4141 cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt
     4201 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4261 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4321 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4381 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4441 aaaaggccgc gttgctggcg tttttccata ggctcggccc ccctgacgag catcacaaaa
     4501 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttcc
     4561 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4621 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     4681 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4741 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4801 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4861 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4921 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4981 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5041 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5101 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5161 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5221 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5281 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5341 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5401 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5461 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5521 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5581 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag
     5641 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5701 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5761 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5821 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5881 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5941 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6001 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6061 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6121 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6181 ttccgcgcac atttccccga aaagtgccac ctgacgtctt attatcatga cattaaccta
     6241 taaaaatagg cgtatcacga ggccctttcg tc
//
