LOCUS       sequence-247338-        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"
     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..3198
                     /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      3220..3407
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3275..3299)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             3820..4623
                     /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(5265..6080)
                     /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(5265..6080)
                     /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     5988..6007
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6072..6091
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6305..6893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6794..6813
                     /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 gagcagcagc ccggggatcc taactcgagg cgaatttctt atgatttatg
     3241 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
     3301 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
     3361 tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat gccgagcaaa
     3421 tgcctgcaaa tcgctcccca tttcgctgga aatctgctcg tcagtggtgc tcacactgac
     3481 gaatcatgta cagatcatac cgatgactgc ctggcgactc acaactaagc aagacagccg
     3541 gaaccagcgc cggcgaacac cactgcatat atggcatatc acaacagtcc acgtctcaag
     3601 cagttacaga gatgttacga accactagtg cactgcagta cacggtttcc ttgaaatttt
     3661 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     3721 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     3781 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     3841 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     3901 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     3961 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     4021 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     4081 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     4141 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     4201 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     4261 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     4321 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     4381 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     4441 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     4501 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     4561 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     4621 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     4681 taattatatc agttattacc cgagtagagc acttgaatcc actgccccgg gaatctcggt
     4741 cgtaatgatt tctataatga cgaaaaaaaa aaaattggaa agaaaaagct tcatggcctt
     4801 tataaaaagg aactatccaa tacctcgcca gaaccaagta acagtatttt acggggcaca
     4861 aatcaagaac aataagacag gactgtaaag atggacgcat tgaactccaa agaacaacaa
     4921 gagttccaaa aagtagtgga acaaaagcaa atgaaggatt tcatgcgttt gtactctaat
     4981 ctggtagaaa gatgtttcac agactgtgtc aatgacttca caacatcaaa gctaaccaat
     5041 aaggaacaaa catgcatcat gaagtgctca gaaaagttct tgaagcatag cgaacgtgta
     5101 gggcagcgtt tccaagaaca aaacgctgcc ttgggacaag gcttgggccg ataaggtgta
     5161 ctggcgtata tatatctaat tatgtatctc tggtgtagcc catttttagc atgtaaatat
     5221 aaagaccgag cggccgcgtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     5281 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     5341 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     5401 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     5461 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     5521 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     5581 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgaggcga
     5641 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     5701 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     5761 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     5821 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     5881 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     5941 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     6001 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctgga gcaagacgtt
     6061 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     6121 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     6181 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagtc atgaccaaaa
     6241 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6301 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6361 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6421 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6481 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6541 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6601 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6661 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6721 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6781 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6841 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6901 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6961 ctgcgttatc ccctgattct gtggataacc gt
//
