LOCUS       sequence-247337-        7328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LEU2 integration vector for Zif268DBD-CRY2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7328)
  AUTHORS   An-Adirekkun JM, Stewart CJ, Geller SH, Patel MT, Melendez J, Oakes 
            BL, Noyes MB, McClean MN
  TITLE     A yeast optogenetic toolkit (yOTK) for gene expression control in 
            Saccharomyces cerevisiae.
  JOURNAL   Biotechnol Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234.
  PUBMED    31788779
REFERENCE   2  (bases 1 to 7328)
  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: "Biotechnol 
            Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234."
COMMENT     Sequence Label: pMM0557
FEATURES             Location/Qualifiers
     source          1..7328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        723..1389
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1259..1282
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1402..1422
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1705..3540
                     /codon_start=1
                     /product="Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAAPDEIVADSFEALGANTIKE
                     PGLCPSVSSNDQQVPSAVRYNGSKRVKPEEEEERDMKKSRGFDERELFSTAESSSSSSV
                     FFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK"
     terminator      3556..3743
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3611..3635)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             4156..4959
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(5601..6416)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5601..6416)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6324..6343
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6408..6427
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6641..7229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7130..7149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc tcatttcatc
       61 catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca agacaagcaa
      121 cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact cagatattga
      181 aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca acggttcatc
      241 atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa ttgaggctac
      301 tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg atgtagaaaa
      361 ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
      421 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
      481 gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt acccctgaat
      541 tcgcatctag actgatgaga cgtggtagag ccacaaacag ccggtacaag caacgatctc
      601 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
      661 cccacagtac agacactgcg acaacgtggc aattcgtcgc aatacaacgc agttcgagtt
      721 tatcattatc aatactgcca tttcaaagaa tacgtaaata attaatagta gtgattttcc
      781 taactttatt tagtcaaaaa attagccttt taattctgct gtaacccgta catgcccaaa
      841 atagggggcg ggttacacag aatatataac atcgtaggtg tctgggtgaa cagtttattc
      901 ctggcatcca ctaaatataa tggagcccgc tttttaagct ggcatccaga aaaaaaaaga
      961 atcccagcac caaaatattg ttttcttcac caaccatcag ttcataggtc cattctctta
     1021 gcgcaactac agagaacagg ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt
     1081 gatgcaacct gcctggagta aatgatgaca caaggcaatt gacccacgca tgtatctatc
     1141 tcattttctt acaccttcta ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa
     1201 aaggttgaaa ccagttccct gaaattattc ccctacttga ctaataagta tataaagacg
     1261 gtaggtattg attgtaattc tgtaaatcta tttcttaaac ttcttaaatt ctacttttat
     1321 agttagtctt ttttttagtt ttaaaacacc aagaacttag tttcgaataa acacacataa
     1381 acaaacaaaa gatctatggc ccccaagaag aagaggaagg ttggtattca cgggggtacc
     1441 cgcccatatg cttgccctgt cgagtcctgc gatcgccgct tttctcgctc ggatgagctt
     1501 acccgccata tccgcatcca taccggtcag aagcccttcc agtgtcgaat ctgcatgcgt
     1561 aacttcagtc gtagtgacca ccttaccacc cacatccgca cccacacagg cgagaagcct
     1621 tttgcctgtg acatttgtgg gaggaagttt gccaggagtg atgaacgcaa gaggcatacc
     1681 aaaatccata caggtgctag cttcatgaag atggacaaaa agactatagt ttggtttaga
     1741 agagatctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
     1801 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
     1861 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
     1921 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
     1981 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
     2041 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
     2101 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
     2161 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
     2221 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
     2281 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggagtcca
     2341 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
     2401 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
     2461 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     2521 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     2581 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     2641 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     2701 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     2761 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     2821 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     2881 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     2941 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     3001 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     3061 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     3121 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
     3181 atcatgatcg gagcagcacc tgatgagatt gtagcagata gcttcgaggc cttaggggct
     3241 aataccatta aagaacctgg tctttgccca tctgtgtctt ctaatgacca acaagtacct
     3301 tcggctgttc gttacaacgg gtcaaagaga gtgaaacctg aggaagaaga agagagagac
     3361 atgaagaaat ctaggggatt cgatgaaagg gagttgtttt cgactgctga atcttcttct
     3421 tcttcgagtg tgtttttcgt ttcgcagtct tgctcgttgg catcagaagg gaagaatctg
     3481 gaaggtattc aagattcatc tgatcagatt actacaagtt tgggaaaaaa tggttgcaaa
     3541 ggatcctaac tcgaggcgaa tttcttatga tttatgattt ttattattaa ataagttata
     3601 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     3661 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct
     3721 cttattgacc acacctctac cggcatgccg agcaaatgcc tgcaaatcgc tccccatttc
     3781 gctggaaatc tgctcgtcag tggtgctcac actgacgaat catgtacaga tcataccgat
     3841 gactgcctgg cgactcacaa ctaagcaaga cagccggaac cagcgccggc gaacaccact
     3901 gcatatatgg catatcacaa cagtccacgt ctcaagcagt tacagagatg ttacgaacca
     3961 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     4021 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     4081 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     4141 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     4201 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     4261 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     4321 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     4381 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     4441 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     4501 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     4561 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     4621 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     4681 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     4741 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     4801 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     4861 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     4921 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     4981 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     5041 tagagcactt gaatccactg ccccgggaat ctcggtcgta atgatttcta taatgacgaa
     5101 aaaaaaaaaa ttggaaagaa aaagcttcat ggcctttata aaaaggaact atccaatacc
     5161 tcgccagaac caagtaacag tattttacgg ggcacaaatc aagaacaata agacaggact
     5221 gtaaagatgg acgcattgaa ctccaaagaa caacaagagt tccaaaaagt agtggaacaa
     5281 aagcaaatga aggatttcat gcgtttgtac tctaatctgg tagaaagatg tttcacagac
     5341 tgtgtcaatg acttcacaac atcaaagcta accaataagg aacaaacatg catcatgaag
     5401 tgctcagaaa agttcttgaa gcatagcgaa cgtgtagggc agcgtttcca agaacaaaac
     5461 gctgccttgg gacaaggctt gggccgataa ggtgtactgg cgtatatata tctaattatg
     5521 tatctctggt gtagcccatt tttagcatgt aaatataaag accgagcggc cgcgtgttac
     5581 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     5641 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     5701 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     5761 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     5821 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     5881 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     5941 ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga
     6001 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     6061 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     6121 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     6181 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     6241 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     6301 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     6361 atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca
     6421 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     6481 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     6541 acttttgctg agttgaagga tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7261 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7321 ataaccgt
//
