LOCUS       sequence-259413-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LEU2 integration vector for Zif268DBD-CRY2PHR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  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 6992)
  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: pMM0558
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     192..215
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             335..355
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             638..2131
                     /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      2153..2340
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2208..2232)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             2753..3556
                     /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(4198..5013)
                     /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(4198..5013)
                     /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     4921..4940
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5005..5024
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5238..5826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5727..5746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6648..6992,1..322)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
ORIGIN
        1 acctcaatgg agtgatgcaa cctgcctgga gtaaatgatg acacaaggca attgacccac
       61 gcatgtatct atctcatttt cttacacctt ctattacctt ctgctctctc tgatttggaa
      121 aaagctgaaa aaaaaggttg aaaccagttc cctgaaatta ttcccctact tgactaataa
      181 gtatataaag acggtaggta ttgattgtaa ttctgtaaat ctatttctta aacttcttaa
      241 attctacttt tatagttagt ctttttttta gttttaaaac accaagaact tagtttcgaa
      301 taaacacaca taaacaaaca aaagatctat ggcccccaag aagaagagga aggttggtat
      361 tcacgggggt acccgcccat atgcttgccc tgtcgagtcc tgcgatcgcc gcttttctcg
      421 ctcggatgag cttacccgcc atatccgcat ccataccggt cagaagccct tccagtgtcg
      481 aatctgcatg cgtaacttca gtcgtagtga ccaccttacc acccacatcc gcacccacac
      541 aggcgagaag ccttttgcct gtgacatttg tgggaggaag tttgccagga gtgatgaacg
      601 caagaggcat accaaaatcc atacaggtgc tagcttcatg aagatggaca aaaagactat
      661 agtttggttt agaagagatc taaggattga ggataatcct gcattagcag cagctgctca
      721 cgaaggatct gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac agttttatcc
      781 tggaagagct tcaagatggt ggatgaaaca atcacttgct cacttatctc aatccttgaa
      841 ggctcttgga tctgacctca ctttaatcaa aacccacaac acgatttcag cgatcttgga
      901 ttgtatccgc gttaccggtg ctacaaaagt cgtctttaac cacctctatg atcctgtttc
      961 gttagttcgg gaccataccg taaaggagaa gctggtggaa cgtgggatct ctgtgcaaag
     1021 ctacaatgga gatctattgt atgaaccgtg ggagatatac tgcgaaaagg gcaaaccttt
     1081 tacgagtttc aattcttact ggaagaaatg cttagatatg tcgattgaat ccgttatgct
     1141 tcctcctcct tggcggttga tgccaataac tgcagcggct gaagcgattt gggcgtgttc
     1201 gattgaagaa ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt tgttaactag
     1261 agcttggagt ccaggatgga gcaatgctga taagttacta aatgagttca tcgagaagca
     1321 gttgatagat tatgcaaaga acagcaagaa agttgttggg aattctactt cactactttc
     1381 tccgtatctc catttcgggg aaataagcgt cagacacgtt ttccagtgtg cccggatgaa
     1441 acaaattata tgggcaagag ataagaacag tgaaggagaa gaaagtgcag atctttttct
     1501 taggggaatc ggtttaagag agtattctcg gtatatatgt ttcaacttcc cgtttactca
     1561 cgagcaatcg ttgttgagtc atcttcggtt tttcccttgg gatgctgatg ttgataagtt
     1621 caaggcctgg agacaaggca ggaccggtta tccgttggtg gatgccggaa tgagagagct
     1681 ttgggctacc ggatggatgc ataacagaat aagagtgatt gtttcaagct ttgctgtgaa
     1741 gtttcttctc cttccatgga aatggggaat gaagtatttc tgggatacac ttttggatgc
     1801 tgatttggaa tgtgacatcc ttggctggca gtatatctct gggagtatcc ccgatggcca
     1861 cgagcttgat cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc cagaaggtga
     1921 gtacataagg caatggcttc ccgagcttgc gagattgcca actgaatgga tccatcatcc
     1981 atgggacgct cctttaaccg tactcaaagc ttctggtgtg gaactcggaa caaactatgc
     2041 gaaacccatt gtagacatcg acacagctcg tgagctacta gctaaagcta tttcaagaac
     2101 ccgtgaagca cagatcatga tcggagcagc agcccgggga tcctaactcg aggcgaattt
     2161 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     2221 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     2281 ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg
     2341 catgccgagc aaatgcctgc aaatcgctcc ccatttcgct ggaaatctgc tcgtcagtgg
     2401 tgctcacact gacgaatcat gtacagatca taccgatgac tgcctggcga ctcacaacta
     2461 agcaagacag ccggaaccag cgccggcgaa caccactgca tatatggcat atcacaacag
     2521 tccacgtctc aagcagttac agagatgtta cgaaccacta gtgcactgca gtacacggtt
     2581 tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     2641 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     2701 attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa
     2761 agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa
     2821 tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga
     2881 attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga
     2941 tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     3001 gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt
     3061 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     3121 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga
     3181 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     3241 atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     3301 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg
     3361 tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt
     3421 ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     3481 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     3541 cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     3601 tagagcttca atttaattat atcagttatt acccgagtag agcacttgaa tccactgccc
     3661 cgggaatctc ggtcgtaatg atttctataa tgacgaaaaa aaaaaaattg gaaagaaaaa
     3721 gcttcatggc ctttataaaa aggaactatc caatacctcg ccagaaccaa gtaacagtat
     3781 tttacggggc acaaatcaag aacaataaga caggactgta aagatggacg cattgaactc
     3841 caaagaacaa caagagttcc aaaaagtagt ggaacaaaag caaatgaagg atttcatgcg
     3901 tttgtactct aatctggtag aaagatgttt cacagactgt gtcaatgact tcacaacatc
     3961 aaagctaacc aataaggaac aaacatgcat catgaagtgc tcagaaaagt tcttgaagca
     4021 tagcgaacgt gtagggcagc gtttccaaga acaaaacgct gccttgggac aaggcttggg
     4081 ccgataaggt gtactggcgt atatatatct aattatgtat ctctggtgta gcccattttt
     4141 agcatgtaaa tataaagacc gagcggccgc gtgttacaac caattaacca attctgatta
     4201 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     4261 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     4321 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     4381 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     4441 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
     4501 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     4561 ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     4621 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     4681 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     4741 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     4801 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     4861 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     4921 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     4981 ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     5041 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     5101 attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     5161 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5881 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc cgccaatgct
     5941 aaattcgagt gaaacacagg aagatcagaa aatcctcatt tcatccatat taacaataat
     6001 ttcaaatgtt tatttgcatt atttgaaact aggcaagaca agcaacgaaa cgtttttgaa
     6061 aattttgagt attttcaata aatttgtaga ggactcagat attgaaaaaa agctacagca
     6121 attaatactt gataagaaga gtattgagaa gggcaacggt tcatcatctc atggatctgc
     6181 acatgaacaa acaccagagt caaacgacgt tgaaattgag gctactgcgc caattgatga
     6241 caatacagac gatgataaca aaccgaagtt atctgatgta gaaaaggatt aaagatgcta
     6301 agagatagtg atgatatttc ataaataatg taattctata tatgttaatt accttttttg
     6361 cgaggcatat ttatggtgaa ggataagttt tgaccatcaa agaaggttaa tgtggctgtg
     6421 gtttcagggt ccataaagcc cacatggata acattacccc tgaattcgca tctagactga
     6481 tgagacgtgg tagagccaca aacagccggt acaagcaacg atctccagga ccatctgaat
     6541 catgcgcgga tgacacgaac tcacgacggc gatcacagac attaacccac agtacagaca
     6601 ctgcgacaac gtggcaattc gtcgcaatac aacgcagttc gagtttatca ttatcaatac
     6661 tgccatttca aagaatacgt aaataattaa tagtagtgat tttcctaact ttatttagtc
     6721 aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta
     6781 cacagaatat ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa
     6841 tataatggag cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa
     6901 tattgttttc ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga
     6961 acaggggcac aaacaggcaa aaaacgggca ca
//
