LOCUS       sequence-90260-e        5943 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Induces Citrine expression from a promoter containing 1 lexA box in 
            yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  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 5943)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: FRP791_insul-(lexA-box)1-PminCYC1-Citrine-TCYC1
FEATURES             Location/Qualifiers
     source          1..5943
                     /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             2575..3291
                     /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      3332..3581
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3348..3366)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3693..3711)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3732..3748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3732..3748)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3745..3767)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3756..3772
                     /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(3780..3786,3787..3804,3805..3810))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3963..3980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4134..4722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4893..5684,5685..5753))
                     /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     5516..5535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5754..5858)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5916..5934
                     /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 ctagagcatg tgctctgtat gtatataaaa
     2461 ctcttgtttt cttcttttct cttctttcct tatacattag gacctttgca gcataaatta
     2521 ctatacttct atactagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatgaga
     2581 tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat
     2641 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa
     2701 ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc
     2761 actactttag gttatggttt gatgtgtttt gctagatacc cagatcatat gaaacaacat
     2821 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     2881 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     2941 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     3001 gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc
     3061 aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat
     3121 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     3181 tcctatcaat ctaagttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     3241 gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa aggatcctaa
     3301 ctcgagacag gccccttttc ctttgtctat atcatgtaat tagttatgtc acgcttacat
     3361 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     3421 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     3481 aaatttttct tttttttctg tacaaacgcg tgtacgcatg taacattata ctgaaaacct
     3541 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caaggttcgc agtttacact
     3601 ctcatcgtcg ctctcatcat cgcttccgtt gttgttttcc ttagtagcgt ctgcttggta
     3661 cccagcttcg gccgtgtacc cagcttttgt tccctttagt gagggttaat tccgagcttg
     3721 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3781 aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc
     3841 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3901 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3961 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4021 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4081 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4141 aggctcggcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4201 ccgacaggac tataaagata ccaggcgttc ccccctggaa gctccctcgt gcgctctcct
     4261 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4321 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4381 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4441 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4501 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4561 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4621 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4681 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4741 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4801 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4861 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4921 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactgcccgt cgtgtagata
     4981 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5041 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5101 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5161 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5221 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5281 gttacatgat cccccatgtt gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5341 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5401 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5461 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5521 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5581 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5641 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5701 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5761 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5821 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5881 cctgacgtct tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5941 gtc
//
