LOCUS       sequence-90262-e        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Induces Citrine expression from a promoter containing 4 lexA boxes 
            in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  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 6084)
  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: FRP793_insul-(lexA-box)4-PminCYC1-Citrine-TCYC1
FEATURES             Location/Qualifiers
     source          1..6084
                     /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             2716..3432
                     /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      3473..3722
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3489..3507)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3834..3852)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3873..3889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3873..3889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3886..3908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3897..3913
                     /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(3921..3927,3928..3945,3946..3951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4104..4121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4275..4863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5034..5825,5826..5894))
                     /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     5657..5676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5895..5999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6057..6075
                     /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 tctagagcat
     2581 gtgctctgta tgtatataaa actcttgttt tcttcttttc tcttctttcc ttatacatta
     2641 ggacctttgc agcataaatt actatacttc tatactagtg gatcccccgg gctgcaggaa
     2701 ttcgatatca agcttatgag atctaaaggt gaagaattat tcactggtgt tgtcccaatt
     2761 ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa
     2821 ggtgatgcta cttacggtaa attgacctta aaatttattt gtactactgg taaattgcca
     2881 gttccatggc caaccttagt cactacttta ggttatggtt tgatgtgttt tgctagatac
     2941 ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg ttatgttcaa
     3001 gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga agtcaagttt
     3061 gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa agaagatggt
     3121 aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta catcatggct
     3181 gacaaacaaa agaatggtat caaagttaac ttcaaaatta gacacaacat tgaagatggt
     3241 tctgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg tccagtcttg
     3301 ttaccagaca accattactt atcctatcaa tctaagttat ccaaagatcc aaacgaaaag
     3361 agagaccaca tggtcttgtt agaatttgtt actgctgctg gtattaccca tggtatggat
     3421 gaattgtaca aaggatccta actcgagaca ggcccctttt cctttgtcta tatcatgtaa
     3481 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     3541 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     3601 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaacgc gtgtacgcat
     3661 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     3721 gcaaggttcg cagtttacac tctcatcgtc gctctcatca tcgcttccgt tgttgttttc
     3781 cttagtagcg tctgcttggt acccagcttc ggccgtgtac ccagcttttg ttccctttag
     3841 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3901 tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt
     3961 gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4021 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4261 gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc
     4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga
     4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4441 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4681 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5041 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5101 tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5161 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5221 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5281 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5341 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5401 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc
     5461 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5521 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5581 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5641 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5701 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5761 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5821 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5881 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5941 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6001 acatttcccc gaaaagtgcc acctgacgtc ttattatcat gacattaacc tataaaaata
     6061 ggcgtatcac gaggcccttt cgtc
//
