LOCUS       sequence-90261-e        5990 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Induces Citrine expression from a promoter containing 2 lexA boxes 
            in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  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 5990)
  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: FRP792_insul-(lexA-box)2-PminCYC1-Citrine-TCYC1
FEATURES             Location/Qualifiers
     source          1..5990
                     /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             2622..3338
                     /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      3379..3628
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3395..3413)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3740..3758)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3779..3795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3779..3795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3792..3814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3803..3819
                     /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(3827..3833,3834..3851,3852..3857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4010..4027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4181..4769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4940..5731,5732..5800))
                     /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     5563..5582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5801..5905)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5963..5981
                     /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 gagcatgtgc tctgtatgta tataaaactc ttgttttctt
     2521 cttttctctt ctttccttat acattaggac ctttgcagca taaattacta tacttctata
     2581 ctagtggatc ccccgggctg caggaattcg atatcaagct tatgagatct aaaggtgaag
     2641 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
     2701 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
     2761 ttatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttaggtt
     2821 atggtttgat gtgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt
     2881 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
     2941 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
     3001 aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata
     3061 actctcacaa tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca
     3121 aaattagaca caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata
     3181 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatcta
     3241 agttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     3301 ctgctggtat tacccatggt atggatgaat tgtacaaagg atcctaactc gagacaggcc
     3361 ccttttcctt tgtctatatc atgtaattag ttatgtcacg cttacattca cgccctcctc
     3421 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     3481 atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     3541 ttttctgtac aaacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     3601 tttgggacgc tcgaaggctt taatttgcaa ggttcgcagt ttacactctc atcgtcgctc
     3661 tcatcatcgc ttccgttgtt gttttcctta gtagcgtctg cttggtaccc agcttcggcc
     3721 gtgtacccag cttttgttcc ctttagtgag ggttaattcc gagcttggcg taatcatggt
     3781 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg
     3841 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt
     3901 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctcggccccc
     4201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4261 aaagatacca ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     4381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4621 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4981 gtctatttcg ttcatccata gttgcctgac tgcccgtcgt gtagataact acgatacggg
     5041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5341 ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcttat
     5941 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
