LOCUS Exported 9089 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast two-hybrid expression vector, expresses Gal4BD fused to Arabidopsis CRY2 (residues 1-535). ACCESSION . VERSION . KEYWORDS Gal4BD-CRY2(535) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9089) AUTHORS Taslimi A, Zoltowski B, Miranda JG, Pathak GP, Hughes RM, Tucker CL TITLE Optimized second-generation CRY2-CIB dimerizers and photoactivatable Cre recombinase. JOURNAL Nat Chem Biol. 2016 Apr 11. doi: 10.1038/nchembio.2063. PUBMED 27065233 REFERENCE 2 (bases 1 to 9089) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9089 /organism="synthetic DNA construct" /mol_type="other DNA" 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 2676..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 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 5332..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 6211..7704 /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 7852..8039 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(7907..7931) /label=pHybLex-R /note="ADH terminator, reverse primer" CDS complement(8092..8766) /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(8767..8868) /gene="S. cerevisiae TRP1" /label=TRP1 promoter primer_bind complement(8883..8899) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8920..8939) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9039..9061 /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 gccggaattc atgaagatgg acaaaaagac tatagtttgg 6241 tttagaagag acctaaggat tgaggataat cctgcattag cagcagctgc tcacgaagga 6301 tctgtttttc ctgtcttcat ttggtgtcct gaagaagaag gacagtttta tcctggaaga 6361 gcttcaagat ggtggatgaa acaatcactt gctcacttat ctcaatcctt gaaggctctt 6421 ggatctgacc tcactttaat caaaacccac aacacgattt cagcgatctt ggattgtatc 6481 cgcgttaccg gtgctacaaa agtcgtcttt aaccacctct atgatcctgt ttcgttagtt 6541 cgggaccata ccgtaaagga gaagctggtg gaacgtggga tctctgtgca aagctacaat 6601 ggagatctat tgtatgaacc gtgggagata tactgcgaaa agggcaaacc ttttacgagt 6661 ttcaattctt actggaagaa atgcttagat atgtcgattg aatccgttat gcttcctcct 6721 ccttggcggt tgatgccaat aactgcagcg gctgaagcga tttgggcgtg ttcgattgaa 6781 gaactagggc tggagaatga ggccgagaaa ccgagcaatg cgttgttaac tagagcttgg 6841 tctccaggat ggagcaatgc tgataagtta ctaaatgagt tcatcgagaa gcagttgata 6901 gattatgcaa agaacagcaa gaaagttgtt gggaattcta cttcactact ttctccgtat 6961 ctccatttcg gggaaataag cgtcagacac gttttccagt gtgcccggat gaaacaaatt 7021 atatgggcaa gagataagaa cagtgaagga gaagaaagtg cagatctttt tcttagggga 7081 atcggtttaa gagagtattc tcggtatata tgtttcaact tcccgtttac tcacgagcaa 7141 tcgttgttga gtcatcttcg gtttttccct tgggatgctg atgttgataa gttcaaggcc 7201 tggagacaag gcaggaccgg ttatccgttg gtggatgccg gaatgagaga gctttgggct 7261 accggatgga tgcataacag aataagagtg attgtttcaa gctttgctgt gaagtttctt 7321 ctccttccat ggaaatgggg aatgaagtat ttctgggata cacttttgga tgctgatttg 7381 gaatgtgaca tccttggctg gcagtatatc tctgggagta tccccgatgg ccacgagctt 7441 gatcgcttgg acaatcccgc gttacaaggc gccaaatatg acccagaagg tgagtacata 7501 aggcaatggc ttcccgagct tgcgagattg ccaactgaat ggatccatca tccatgggac 7561 gctcctttaa ccgtactcaa agcttctggt gtggaactcg gaacaaacta tgcgaaaccc 7621 attgtagaca tcgacacagc tcgtgagcta ctagctaaag ctatttcaag aacccgtgaa 7681 gcacagatca tgatcggagc agcacctgat gagattgtag cagatagctt cgaggcctta 7741 ggggctaata ccattaaaga acctggtctt tgcccatctg tgtcttctaa tgaccaacaa 7801 gtaccttcgg ctgtttgagg atccgtcgac ctgcagccaa gctaattccg ggcgaatttc 7861 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt 7921 taaagtgact cttaggtttt aaaacgaaaa ttcttgttct tgagtaactc tttcctgtag 7981 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc 8041 atgccggcaa gtgcacaaac aatacttaaa taaatactac tcagtaataa cctatttctt 8101 agcatttttg acgaaatttg ctattttgtt agagtctttt acaccatttg tctccacacc 8161 tccgcttaca tcaacaccaa taacgccatt taatctaagc gcatcaccaa cattttctgg 8221 cgtcagtcca ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca 8281 gtcagaaatc gagttccaat ccaaaagttc acctgtccca cctgcttctg aatcaaacaa 8341 gggaataaac gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc agtcttttgg 8401 aaatacgagt cttttaataa ctggcaaacc gaggaactct tggtattctt gccacgactc 8461 atctccatgc agttggacga tatcaatgcc gtaatcattg accagagcca aaacatcctc 8521 cttaggttga ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac tatttttata 8581 tgcttttaca agacttgaaa ttttccttgc aataaccggg tcaattgttc tctttctatt 8641 gggcacacat ataataccca gcaagtcagc atcggaatct agagcacatt ctgcggcctc 8701 tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga aattaataac 8761 agacatactc caagctgcct ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca 8821 ccaatgccct ccctcttggc cctctccttt tcttttttcg accgaattaa ttcgtaatca 8881 tggtcatagc tgtttcctgt cgatatggtg cactctcagt acaatctgct ctgatgccgc 8941 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 9001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 9061 gttttcaccg tcatcaccga aacgcgcga //