LOCUS       sequence-259414-        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        1..667
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     537..560
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             680..700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             983..2818
                     /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      2834..3021
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2889..2913)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             3434..4237
                     /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(4879..5694)
                     /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(4879..5694)
                     /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     5602..5621
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5686..5705
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5919..6507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6408..6427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
       61 actttattta gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat
      121 agggggcggg ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct
      181 ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat
      241 cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc
      301 gcaactacag agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga
      361 tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc
      421 attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa
      481 ggttgaaacc agttccctga aattattccc ctacttgact aataagtata taaagacggt
      541 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
      601 ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgaataaac acacataaac
      661 aaacaaaaga tctatggccc ccaagaagaa gaggaaggtt ggtattcacg ggggtacccg
      721 cccatatgct tgccctgtcg agtcctgcga tcgccgcttt tctcgctcgg atgagcttac
      781 ccgccatatc cgcatccata ccggtcagaa gcccttccag tgtcgaatct gcatgcgtaa
      841 cttcagtcgt agtgaccacc ttaccaccca catccgcacc cacacaggcg agaagccttt
      901 tgcctgtgac atttgtggga ggaagtttgc caggagtgat gaacgcaaga ggcataccaa
      961 aatccataca ggtgctagct tcatgaagat ggacaaaaag actatagttt ggtttagaag
     1021 agatctaagg attgaggata atcctgcatt agcagcagct gctcacgaag gatctgtttt
     1081 tcctgtcttc atttggtgtc ctgaagaaga aggacagttt tatcctggaa gagcttcaag
     1141 atggtggatg aaacaatcac ttgctcactt atctcaatcc ttgaaggctc ttggatctga
     1201 cctcacttta atcaaaaccc acaacacgat ttcagcgatc ttggattgta tccgcgttac
     1261 cggtgctaca aaagtcgtct ttaaccacct ctatgatcct gtttcgttag ttcgggacca
     1321 taccgtaaag gagaagctgg tggaacgtgg gatctctgtg caaagctaca atggagatct
     1381 attgtatgaa ccgtgggaga tatactgcga aaagggcaaa ccttttacga gtttcaattc
     1441 ttactggaag aaatgcttag atatgtcgat tgaatccgtt atgcttcctc ctccttggcg
     1501 gttgatgcca ataactgcag cggctgaagc gatttgggcg tgttcgattg aagaactagg
     1561 gctggagaat gaggccgaga aaccgagcaa tgcgttgtta actagagctt ggagtccagg
     1621 atggagcaat gctgataagt tactaaatga gttcatcgag aagcagttga tagattatgc
     1681 aaagaacagc aagaaagttg ttgggaattc tacttcacta ctttctccgt atctccattt
     1741 cggggaaata agcgtcagac acgttttcca gtgtgcccgg atgaaacaaa ttatatgggc
     1801 aagagataag aacagtgaag gagaagaaag tgcagatctt tttcttaggg gaatcggttt
     1861 aagagagtat tctcggtata tatgtttcaa cttcccgttt actcacgagc aatcgttgtt
     1921 gagtcatctt cggtttttcc cttgggatgc tgatgttgat aagttcaagg cctggagaca
     1981 aggcaggacc ggttatccgt tggtggatgc cggaatgaga gagctttggg ctaccggatg
     2041 gatgcataac agaataagag tgattgtttc aagctttgct gtgaagtttc ttctccttcc
     2101 atggaaatgg ggaatgaagt atttctggga tacacttttg gatgctgatt tggaatgtga
     2161 catccttggc tggcagtata tctctgggag tatccccgat ggccacgagc ttgatcgctt
     2221 ggacaatccc gcgttacaag gcgccaaata tgacccagaa ggtgagtaca taaggcaatg
     2281 gcttcccgag cttgcgagat tgccaactga atggatccat catccatggg acgctccttt
     2341 aaccgtactc aaagcttctg gtgtggaact cggaacaaac tatgcgaaac ccattgtaga
     2401 catcgacaca gctcgtgagc tactagctaa agctatttca agaacccgtg aagcacagat
     2461 catgatcgga gcagcacctg atgagattgt agcagatagc ttcgaggcct taggggctaa
     2521 taccattaaa gaacctggtc tttgcccatc tgtgtcttct aatgaccaac aagtaccttc
     2581 ggctgttcgt tacaacgggt caaagagagt gaaacctgag gaagaagaag agagagacat
     2641 gaagaaatct aggggattcg atgaaaggga gttgttttcg actgctgaat cttcttcttc
     2701 ttcgagtgtg tttttcgttt cgcagtcttg ctcgttggca tcagaaggga agaatctgga
     2761 aggtattcaa gattcatctg atcagattac tacaagtttg ggaaaaaatg gttgcaaagg
     2821 atcctaactc gaggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     2881 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     2941 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct
     3001 tattgaccac acctctaccg gcatgccgag caaatgcctg caaatcgctc cccatttcgc
     3061 tggaaatctg ctcgtcagtg gtgctcacac tgacgaatca tgtacagatc ataccgatga
     3121 ctgcctggcg actcacaact aagcaagaca gccggaacca gcgccggcga acaccactgc
     3181 atatatggca tatcacaaca gtccacgtct caagcagtta cagagatgtt acgaaccact
     3241 agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa
     3301 ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga
     3361 agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa
     3421 acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc
     3481 tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt
     3541 ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg
     3601 tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc
     3661 gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga
     3721 cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc
     3781 agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc
     3841 ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa
     3901 gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa
     3961 agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga
     4021 ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca
     4081 gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg
     4141 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
     4201 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
     4261 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta
     4321 gagcacttga atccactgcc ccgggaatct cggtcgtaat gatttctata atgacgaaaa
     4381 aaaaaaaatt ggaaagaaaa agcttcatgg cctttataaa aaggaactat ccaatacctc
     4441 gccagaacca agtaacagta ttttacgggg cacaaatcaa gaacaataag acaggactgt
     4501 aaagatggac gcattgaact ccaaagaaca acaagagttc caaaaagtag tggaacaaaa
     4561 gcaaatgaag gatttcatgc gtttgtactc taatctggta gaaagatgtt tcacagactg
     4621 tgtcaatgac ttcacaacat caaagctaac caataaggaa caaacatgca tcatgaagtg
     4681 ctcagaaaag ttcttgaagc atagcgaacg tgtagggcag cgtttccaag aacaaaacgc
     4741 tgccttggga caaggcttgg gccgataagg tgtactggcg tatatatatc taattatgta
     4801 tctctggtgt agcccatttt tagcatgtaa atataaagac cgagcggccg cgtgttacaa
     4861 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     4921 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     4981 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     5041 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     5101 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     5161 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     5221 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
     5281 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     5341 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     5401 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     5461 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     5521 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     5581 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     5641 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     5701 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     5761 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     5821 ttttgctgag ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6061 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6601 aaccgtgcgg ccgccaatgc taaattcgag tgaaacacag gaagatcaga aaatcctcat
     6661 ttcatccata ttaacaataa tttcaaatgt ttatttgcat tatttgaaac taggcaagac
     6721 aagcaacgaa acgtttttga aaattttgag tattttcaat aaatttgtag aggactcaga
     6781 tattgaaaaa aagctacagc aattaatact tgataagaag agtattgaga agggcaacgg
     6841 ttcatcatct catggatctg cacatgaaca aacaccagag tcaaacgacg ttgaaattga
     6901 ggctactgcg ccaattgatg acaatacaga cgatgataac aaaccgaagt tatctgatgt
     6961 agaaaaggat taaagatgct aagagatagt gatgatattt cataaataat gtaattctat
     7021 atatgttaat tacctttttt gcgaggcata tttatggtga aggataagtt ttgaccatca
     7081 aagaaggtta atgtggctgt ggtttcaggg tccataaagc ccacatggat aacattaccc
     7141 ctgaattcgc atctagactg atgagacgtg gtagagccac aaacagccgg tacaagcaac
     7201 gatctccagg accatctgaa tcatgcgcgg atgacacgaa ctcacgacgg cgatcacaga
     7261 cattaaccca cagtacagac actgcgacaa cgtggcaatt cgtcgcaata caacgcagtt
     7321 cgagttta
//
