LOCUS       Exported                9088 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 9088)
  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 9088)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9088
                     /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(8882..8898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8919..8938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9038..9060
                     /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 gctgtcttgc 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 tgtcatagct gtttcctgtc gatatggtgc actctcagta caatctgctc tgatgccgca
     8941 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     9001 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     9061 ttttcaccgt catcaccgaa acgcgcga
//
