LOCUS       sequence-259442-        9147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for optogenetic machinery 
            (pSTRONG-VP16-CIB1/pMEDIUM-ZCRY2PHR) integration effects.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9147)
  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 9147)
  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: pMM0648
FEATURES             Location/Qualifiers
     source          1..9147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        228..641
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             691..711
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     939..958
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        939..957
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             982..1491
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             3011..3031
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3314..4807
                     /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"
     CDS             5349..6152
                     /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(6794..7609)
                     /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(6794..7609)
                     /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     7517..7536
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7601..7620
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7834..8422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8323..8342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attattgaca gcctagacat caatagtcat acaacagaaa gcgaccaccc aactttggct
       61 gataatagcg tataaacaat gcatactttg tacgttcaaa atacaatgca gtagatatat
      121 ttatgcatat tacatataat acatatcaca taggaagcaa caggcgcgtt ggacttttaa
      181 ttttcgagga ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat cccccacaca
      241 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
      301 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
      361 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga caccgcctcg
      421 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
      481 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa
      541 cggtcatcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac
      601 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaagatct
      661 atggataaag cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt
      721 accgccgcct cggagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg
      781 ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc
      841 ccccacgact ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg
      901 tttaccgatg cccttggaat tgacgagtac ggtgggttta atacgactca ctatagggcg
      961 agcgccgaag ctagcgccac catgaatgga gctataggag gtgacctttt gctcaatttt
     1021 cctgacatgt cggtcctaga gcgccaaagg gctcacctca agtacctcaa tcccaccttt
     1081 gattctcctc tcgccggctt ctttgccgat tcttcaatga ttaccggcgg cgagatggac
     1141 agctatcttt cgactgccgg tttgaatctt ccgatgatgt acggtgaaac gacggtggaa
     1201 ggtgattcaa gactctcaat ttcgccggaa acgacgcttg ggactggaaa tttcaagaaa
     1261 cggaagtttg atacagagac taaggattgt aatgagaaga agaagaagat gacgatgaac
     1321 agagatgacc tagtagaaga aggagaagaa gagaagtcga aaataacaga gcaaaacaat
     1381 gggagcacaa aaagcatcaa gaagatgaaa cacaaagcca agaaagaaga gaacaatttc
     1441 tctaatgatt catctaaagt gacgaaggaa ttggagaaaa cggattatat tcatgttcgt
     1501 gcacgacgag gccaagccac tgatagtcac agcatagcag aacgagttag aagagaaaag
     1561 atcagtgaga gaatgaagtt tctacaagat ttggttcctg gatgcgacaa gatcacaggc
     1621 aaagcaggga tgcttgatga aatcattaac tatgttcagt ctcttcagag acaaatcgag
     1681 ttcttatcga tgaaactagc aattgtgaat ccaaggccgg attttgatat ggatgacatt
     1741 tttgccaaag aggttgcctc aactccaatg actgtggtgc catctcctga aatggttctt
     1801 tccggttatt ctcatgagat ggttcactct ggttattcta gtgagatggt taactccggt
     1861 taccttcatg tcaatccaat gcagcaagtg aataccagtt ctgatccatt gtcatgcttc
     1921 aacaatggcg aagctccttc gatgtgggac tctcatgtgc agaatctcta tggcaattta
     1981 ggagtaccgg tcatcgagct cgagctgcag atgaatcgta gatacggatc ctaactcgag
     2041 agcttttgat taagccttct agtccaaaaa acacgttttt ttgtcattta tttcattttc
     2101 ttagaatagt ttagtttatt cattttatag tcacgaatgt tttatgattc tatatagggt
     2161 tgcaaacaag catttttcat tttatgttaa aacaatttca ggtttacctt ttattctgct
     2221 tgtggtgacg cgtgtatccg cccgctcttt tggtcaccca tgtatgctgc caaaccagat
     2281 gtcaacacag ctacaacgaa gaggatgtcc aatatttttt ttaaggaata aggatacttc
     2341 aagactagat tcccccctgc attcccatca gaaccgtaaa ccttggcgct ttccttggga
     2401 agtattcaag aagtgccttg tccggtttct gtggctcaca aaccagcgcg cccgatatgg
     2461 ctttcttttc acttatgaat gtaccagtac gggacaatta gaacgctcct gtaacaatct
     2521 ctttgcaaat gtggggttac attctaacca tgtcacactg ctgacgaaat tcaaagtaaa
     2581 aaaaaatggg accacgtctt gagaacgata gattttcttt attttacatt gaacagtcgt
     2641 tgtctcagcg cgctttatgt tttcattcat acttcatatt ataaaataac aaaagaagaa
     2701 tttcatattc acgcccaaga aatcaggctg ctttccaaat gcaattgaca cttcattagc
     2761 catcacacaa aactctttct tgctggagct tcttttaaaa aagacctcag tacaccaaac
     2821 acgttacccg acctcgttat tttacgacaa ctatgataaa attctgaaga aaaaataaaa
     2881 aaattttcat acttcttgct tttatttaaa ccattgaatg atttcttttg aacaaaacta
     2941 cctgtttcac caaaggaaat agaaagaaaa aatcaattag aagaaaacaa aaaacaaaag
     3001 atctatggcc cccaagaaga agaggaaggt tggtattcac gggggtaccc gcccatatgc
     3061 ttgccctgtc gagtcctgcg atcgccgctt ttctcgctcg gatgagctta cccgccatat
     3121 ccgcatccat accggtcaga agcccttcca gtgtcgaatc tgcatgcgta acttcagtcg
     3181 tagtgaccac cttaccaccc acatccgcac ccacacaggc gagaagcctt ttgcctgtga
     3241 catttgtggg aggaagtttg ccaggagtga tgaacgcaag aggcatacca aaatccatac
     3301 aggtgctagc ttcatgaaga tggacaaaaa gactatagtt tggtttagaa gagatctaag
     3361 gattgaggat aatcctgcat tagcagcagc tgctcacgaa ggatctgttt ttcctgtctt
     3421 catttggtgt cctgaagaag aaggacagtt ttatcctgga agagcttcaa gatggtggat
     3481 gaaacaatca cttgctcact tatctcaatc cttgaaggct cttggatctg acctcacttt
     3541 aatcaaaacc cacaacacga tttcagcgat cttggattgt atccgcgtta ccggtgctac
     3601 aaaagtcgtc tttaaccacc tctatgatcc tgtttcgtta gttcgggacc ataccgtaaa
     3661 ggagaagctg gtggaacgtg ggatctctgt gcaaagctac aatggagatc tattgtatga
     3721 accgtgggag atatactgcg aaaagggcaa accttttacg agtttcaatt cttactggaa
     3781 gaaatgctta gatatgtcga ttgaatccgt tatgcttcct cctccttggc ggttgatgcc
     3841 aataactgca gcggctgaag cgatttgggc gtgttcgatt gaagaactag ggctggagaa
     3901 tgaggccgag aaaccgagca atgcgttgtt aactagagct tggagtccag gatggagcaa
     3961 tgctgataag ttactaaatg agttcatcga gaagcagttg atagattatg caaagaacag
     4021 caagaaagtt gttgggaatt ctacttcact actttctccg tatctccatt tcggggaaat
     4081 aagcgtcaga cacgttttcc agtgtgcccg gatgaaacaa attatatggg caagagataa
     4141 gaacagtgaa ggagaagaaa gtgcagatct ttttcttagg ggaatcggtt taagagagta
     4201 ttctcggtat atatgtttca acttcccgtt tactcacgag caatcgttgt tgagtcatct
     4261 tcggtttttc ccttgggatg ctgatgttga taagttcaag gcctggagac aaggcaggac
     4321 cggttatccg ttggtggatg ccggaatgag agagctttgg gctaccggat ggatgcataa
     4381 cagaataaga gtgattgttt caagctttgc tgtgaagttt cttctccttc catggaaatg
     4441 gggaatgaag tatttctggg atacactttt ggatgctgat ttggaatgtg acatccttgg
     4501 ctggcagtat atctctggga gtatccccga tggccacgag cttgatcgct tggacaatcc
     4561 cgcgttacaa ggcgccaaat atgacccaga aggtgagtac ataaggcaat ggcttcccga
     4621 gcttgcgaga ttgccaactg aatggatcca tcatccatgg gacgctcctt taaccgtact
     4681 caaagcttct ggtgtggaac tcggaacaaa ctatgcgaaa cccattgtag acatcgacac
     4741 agctcgtgag ctactagcta aagctatttc aagaacccgt gaagcacaga tcatgatcgg
     4801 agcagcagcc cggggatcct aactcgaggc caattggtgc ggcaattgat aataacgaaa
     4861 atgtctttta atgatctggg tataatgagg aattttccga acgtttttac tttatatata
     4921 tatatacatg taacatatat tctatacgct atagagaaag gaaatttttc aattaaaaaa
     4981 aaaatagaga aagagtttca cttcttgatt atcgctaaca ctaatggttg aagtactgct
     5041 actttaattt tatgctggat gcacacactg gcttaagatg acaacgacta gagatctatg
     5101 tgaggatcct aactcgagat cgagctgagc agttacagag atgttacgaa ccactagtgc
     5161 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
     5221 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     5281 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
     5341 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     5401 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     5461 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     5521 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     5581 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     5641 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     5701 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     5761 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     5821 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     5881 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     5941 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     6001 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     6061 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     6121 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     6181 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtagagca
     6241 cttgaatcca ctgccccggg aatctcggtc gtaatgattt ctataatgac gaaaaaaaaa
     6301 aaattggaaa gaaaaagctt catggccttt ataaaaagga actatccaat acctcgccag
     6361 aaccaagtaa cagtatttta cggggcacaa atcaagaaca ataagacagg actgtaaaga
     6421 tggacgcatt gaactccaaa gaacaacaag agttccaaaa agtagtggaa caaaagcaaa
     6481 tgaaggattt catgcgtttg tactctaatc tggtagaaag atgtttcaca gactgtgtca
     6541 atgacttcac aacatcaaag ctaaccaata aggaacaaac atgcatcatg aagtgctcag
     6601 aaaagttctt gaagcatagc gaacgtgtag ggcagcgttt ccaagaacaa aacgctgcct
     6661 tgggacaagg cttgggccga taaggtgtac tggcgtatat atatctaatt atgtatctct
     6721 ggtgtagccc atttttagca tgtaaatata aagaccgagc ggccgcgtgt tacaaccaat
     6781 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     6841 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     6901 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     6961 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     7021 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     7081 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     7141 tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac
     7201 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     7261 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     7321 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     7381 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     7441 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     7501 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     7561 aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg
     7621 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     7681 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     7741 ctgagttgaa ggatcagtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7801 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7861 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7921 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7981 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8341 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8401 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8461 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8521 tgcggccgcc aatgctaaat tcgagtgaaa cacaggaaga tcagaaaatc ctcatttcat
     8581 ccatattaac aataatttca aatgtttatt tgcattattt gaaactaggc aagacaagca
     8641 acgaaacgtt tttgaaaatt ttgagtattt tcaataaatt tgtagaggac tcagatattg
     8701 aaaaaaagct acagcaatta atacttgata agaagagtat tgagaagggc aacggttcat
     8761 catctcatgg atctgcacat gaacaaacac cagagtcaaa cgacgttgaa attgaggcta
     8821 ctgcgccaat tgatgacaat acagacgatg ataacaaacc gaagttatct gatgtagaaa
     8881 aggattaaag atgctaagag atagtgatga tatttcataa ataatgtaat tctatatatg
     8941 ttaattacct tttttgcgag gcatatttat ggtgaaggat aagttttgac catcaaagaa
     9001 ggttaatgtg gctgtggttt cagggtccat aaagcccaca tggataacat tacccctgaa
     9061 ttcgcatcta gactgaactg gccgataatt gcagacgaac gccttgccaa cagggagttc
     9121 ttcagagaca tggaggctca aaacgaa
//
