LOCUS       sequence-13374-e        8981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gal4 DNA Binding Domain fused to CRY2PHR (aa1-498) for 
            blue-light-induced transcription activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8981)
  AUTHORS   Kennedy MJ, Hughes RM, Peteya LA, Schwartz JW, Ehlers MD, Tucker CL
  TITLE     Rapid blue-light-mediated induction of protein interactions in 
            living cells.
  JOURNAL   Nat Methods. 2010 Dec;7(12):973-5. doi: 10.1038/nmeth.1524
  PUBMED    21037589
REFERENCE   2  (bases 1 to 8981)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2010-12"; volume: "7"; issue: 
            "12"; pages: "973-5"
COMMENT     Sequence Label: pGal4BD-CRY2PHR
FEATURES             Location/Qualifiers
     source          1..8981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(801..820)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             812..1627
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCWRPRLNSNMDADLYGYKWARDNVGQSGATIFRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIRGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHTLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(885..904)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1764..2352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2506..2523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2676..2681,2682..2699,2700..2706)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2714..2730
                     /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)."
     primer_bind     2719..2741
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2738..2754
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2775..2793
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(2824..3071)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3037..3055
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        5029..5733
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5761..6201
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             6217..7710
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     terminator      7744..7931
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(7799..7823)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(7984..8658)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(8659..8760)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8775..8791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8812..8831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8931..8953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatctg cagctctggc ccgtgtctca
      721 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
      781 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
      841 ctggaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      901 cgataatgtc gggcaatcag gtgcgacaat ctttcgattg tatgggaagc ccgatgcgcc
      961 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1021 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1081 tcctgatgat gcatggttac tcaccactgc gatccgcggg aaaacagcat tccaggtatt
     1141 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1201 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1261 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1321 taatggctgg cctgttgaac aagtctggaa agaaatgcat acgcttttgc cattctcacc
     1381 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1441 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1501 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1561 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1621 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     1681 ggacggcgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1741 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1801 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1861 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1921 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1981 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2041 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2101 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     2161 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2221 ggaacaggag agcgcacgag ggagcttcca ggggggaacg cctggtatct ttatagtcct
     2281 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggccg
     2341 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2401 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2461 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2521 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2581 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2641 aatgtgagtt acctcactca ttaggcaccc caggctttac actttatgct tccggctcct
     2701 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     2761 tacgccaagc tcggaattaa ccctcactaa agggaacaaa agctggtacc gatcccgagc
     2821 tttgcaaatt aaagccttcg agcgtcccaa aaccttctca agcaaggttt tcagtataat
     2881 gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata taaataacgt
     2941 tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag gttgtctaac
     3001 tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc gtgacataac
     3061 taattacatg atatcgacaa aggaaaaggg gcctgtttac tcacaggctt ttttcaagta
     3121 ggtaattaag tcgtttctgt ctttttcctt cttcaaccca ccaaaggcca tcttggtact
     3181 tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt
     3241 tttttttttt tttttttttt tttttttttt tttcatagaa ataatacaga agtagatgtt
     3301 gaattagatt aaactgaaga tatataattt attggaaaat acatagagct ttttgttgat
     3361 gcgcttaagc gatcaattca acaacaccac cagcagctct gattttttct tcagccaact
     3421 tggagacgaa tctagctttg acgataactg gaacatttgg aattctaccc ttacccaaga
     3481 tcttaccgta accggctgcc aaagtgtcaa taactggagc agtttcctta gaagcagatt
     3541 tcaagtattg gtctctcttg tcttctggga tcaatgtcca caatttgtcc aagttcaaga
     3601 ctggcttcca gaaatgagct tgttgcttgt ggaagtatct cataccaacc ttaccgaaat
     3661 aacctggatg gtatttatcc atgttaattc tgtggtgatg ttgaccaccg gccatacctc
     3721 taccaccggg gtgctttctg tgcttaccga tacgaccttt accggctgag acgtgacctc
     3781 tgtgctttct agtcttagtg aatctggaag gcattcttga ttagttggat gattgttctg
     3841 ggatttaatg caaaaatcac ttaagaagga aaatcaacgg agaaagcaaa cgccatctta
     3901 aatatacggg atacagatga aagggtttga acctatctgg aaaatagcat taaacaagcg
     3961 aaaaactgcg aggaaaattg tttgcgtctc tgcgggctat tcacgcgcca gaggaaaata
     4021 ggaaaaataa cagggcatta gaaaaataat tttgattttg gtaatgtgtg ggtcctggtg
     4081 tacagatgtt acattggtta cagtactctt gtttttgctg tgtttttcga tgaatctcca
     4141 aaatggttgt tagcacatgg aagagtcacc gatgctaagt tatctctatg taagctacgt
     4201 ggcgtgactt ttgatgaagc cgcacaagag atacaggatt ggcaactgca aatagaatct
     4261 ggggatcccc cctcgagatc cgggatcgaa gaaatgatgg taaatgaaat aggaaatcaa
     4321 ggagcatgaa ggcaaaagac aaatataagg gtcgaacgaa aaataaagtg aaaagtgttg
     4381 atatgatgta tttggctttg cggcgccgaa aaaacgagtt tacgcaattg cacaatcatg
     4441 ctgactctgt ggcggacccg cgctcttgcc ggcccggcga taacgctggg cgtgaggctg
     4501 tgcccggcgg agttttttgc gcctgcattt tccaaggttt accctgcgct aaggggcgag
     4561 attggagaag caataagaat gccggttggg gttgcgatga tgacgaccac gacaactggt
     4621 gtcattattt aagttgccga aagaacctga gtgcatttgc aacatgagta tactagaaga
     4681 atgagccaag acttgcgaga cgcgagtttg ccggtggtgc gaacaataga gcgaccatga
     4741 ccttgaaggt gagacgcgca taaccgctag agtactttga agaggaaaca gcaatagggt
     4801 tgctaccagt ataaatagac aggtacatac aacactggaa atggttgtct gtttgagtac
     4861 gctttcaatt catttgggtg tgcactttat tatgttacaa tatggaaggg aactttacac
     4921 ttctcctatg cacatatatt aattaaagtc caatgctagt agagaagggg ggtaacaccc
     4981 ctccgcgctc ttttccgatt tttttctaaa ccgtggaata tttcggatat ccttttgttg
     5041 tttccgggtg tacaatatgg acttcctctt ttctggcaac caaacccata catcgggatt
     5101 cctataatac cttcgttggt ctccctaaca tgtaggtggc ggaggggaga tatacaatag
     5161 aacagatacc agacaagaca taatgggcta aacaagacta caccaattac actgcctcat
     5221 tgatggtggt acataacgaa ctaatactgt agccctagac ttgatagcca tcatcatatc
     5281 gaagtttcac tacccttttt ccatttgcca tctattgaag taataatagg cgcatgcaac
     5341 ttcttttctt tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga
     5401 caaaaaaaat gatggaagac actaaaggaa aaaattaacg acaaagacag caccaacaga
     5461 tgtcgttgtt ccagagctga tgaggggtat cttcgaacac acgaaacttt ttccttcctt
     5521 cattcacgca cactactctc taatgagcaa cggtatacgg ccttccttcc agttacttga
     5581 atttgaaata aaaaaagttt gccgctttgc tatcaagtat aaatagacct gcaattatta
     5641 atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca
     5701 atatttcaag ctataccaag catacaatca actccaagct tgaagcaagc ctcctgaaag
     5761 atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag
     5821 tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac
     5881 tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg
     5941 ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct tgacatgatt
     6001 ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt tgtacaagat
     6061 aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga tatgcctcta
     6121 acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag taacaaaggt
     6181 caaagacagt tgactgtatc gccgctcgag gccaccatga agatggacaa aaagactata
     6241 gtttggttta gaagagacct aaggattgag gataatcctg cattagcagc agctgctcac
     6301 gaaggatctg tttttcctgt cttcatttgg tgtcctgaag aagaaggaca gttttatcct
     6361 ggaagagctt caagatggtg gatgaaacaa tcacttgctc acttatctca atccttgaag
     6421 gctcttggat ctgacctcac tttaatcaaa acccacaaca cgatttcagc gatcttggat
     6481 tgtatccgcg ttaccggtgc tacaaaagtc gtctttaacc acctctatga tcctgtttcg
     6541 ttagttcggg accataccgt aaaggagaag ctggtggaac gtgggatctc tgtgcaaagc
     6601 tacaatggag atctattgta tgaaccgtgg gagatatact gcgaaaaggg caaacctttt
     6661 acgagtttca attcttactg gaagaaatgc ttagatatgt cgattgaatc cgttatgctt
     6721 cctcctcctt ggcggttgat gccaataact gcagcggctg aagcgatttg ggcgtgttcg
     6781 attgaagaac tagggctgga gaatgaggcc gagaaaccga gcaatgcgtt gttaactaga
     6841 gcttggtctc caggatggag caatgctgat aagttactaa atgagttcat cgagaagcag
     6901 ttgatagatt atgcaaagaa cagcaagaaa gttgttggga attctacttc actactttct
     6961 ccgtatctcc atttcgggga aataagcgtc agacacgttt tccagtgtgc ccggatgaaa
     7021 caaattatat gggcaagaga taagaacagt gaaggagaag aaagtgcaga tctttttctt
     7081 aggggaatcg gtttaagaga gtattctcgg tatatatgtt tcaacttccc gtttactcac
     7141 gagcaatcgt tgttgagtca tcttcggttt ttcccttggg atgctgatgt tgataagttc
     7201 aaggcctgga gacaaggcag gaccggttat ccgttggtgg atgccggaat gagagagctt
     7261 tgggctaccg gatggatgca taacagaata agagtgattg tttcaagctt tgctgtgaag
     7321 tttcttctcc ttccatggaa atggggaatg aagtatttct gggatacact tttggatgct
     7381 gatttggaat gtgacatcct tggctggcag tatatctctg ggagtatccc cgatggccac
     7441 gagcttgatc gcttggacaa tcccgcgtta caaggcgcca aatatgaccc agaaggtgag
     7501 tacataaggc aatggcttcc cgagcttgcg agattgccaa ctgaatggat ccatcatcca
     7561 tgggacgctc ctttaaccgt actcaaagct tctggtgtgg aactcggaac aaactatgcg
     7621 aaacccattg tagacatcga cacagctcgt gagctactag ctaaagctat ttcaagaacc
     7681 cgtgaagcac agatcatgat cggagcagca gcccgggtcg acctgcagcc aagctaattc
     7741 cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     7801 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttgtt cttgagtaac
     7861 tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac
     7921 acctctaccg gcatgccggc aagtgcacaa acaatactta aataaatact actcagtaat
     7981 aacctatttc ttagcatttt tgacgaaatt tgctattttg ttagagtctt ttacaccatt
     8041 tgtctccaca cctccgctta catcaacacc aataacgcca tttaatctaa gcgcatcacc
     8101 aacattttct ggcgtcagtc caccagctaa cataaaatgt aagctttcgg ggctctcttg
     8161 ccttccaacc cagtcagaaa tcgagttcca atccaaaagt tcacctgtcc cacctgcttc
     8221 tgaatcaaac aagggaataa acgaatgagg tttctgtgaa gctgcactga gtagtatgtt
     8281 gcagtctttt ggaaatacga gtcttttaat aactggcaaa ccgaggaact cttggtattc
     8341 ttgccacgac tcatctccat gcagttggac gatatcaatg ccgtaatcat tgaccagagc
     8401 caaaacatcc tccttaggtt gattacgaaa cacgccaacc aagtatttcg gagtgcctga
     8461 actattttta tatgctttta caagacttga aattttcctt gcaataaccg ggtcaattgt
     8521 tctctttcta ttgggcacac atataatacc cagcaagtca gcatcggaat ctagagcaca
     8581 ttctgcggcc tctgtgctct gcaagccgca aactttcacc aatggaccag aactacctgt
     8641 gaaattaata acagacatac tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt
     8701 gctcaatagt caccaatgcc ctccctcttg gccctctcct tttctttttt cgaccgaatt
     8761 aattcgtaat catggtcata gctgtttcct gtcgatatgg tgcactctca gtacaatctg
     8821 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     8881 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     8941 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg a
//
