LOCUS       sequence-258775-        7153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for ZIF268DBD-L3-CRY2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  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 7153)
  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: pMM0521
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             238..2076
                     /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      2110..2297
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2165..2189)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2320..2340)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2320..2338)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2359..2375)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2359..2375)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2372..2394)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2383..2399
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2407..2413,2414..2431,2432..2437))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2452..2473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2590..2607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2761..3349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2841..2860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3520..4311,4312..4380))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4143..4162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4381..4485)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4522..5025
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5066..5084
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5122..5144)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5244..5263
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5284..5564
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5565..6239
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(6479..6500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6673..7069
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             7088..7108
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tgcgatcgcc gcttttctcg ctcggatgag cttacccgcc atatccgcat ccataccggt
       61 cagaagccct tccagtgtcg aatctgcatg cgtaacttca gtcgtagtga ccaccttacc
      121 acccacatcc gcacccacac aggcgagaag ccttttgcct gtgacatttg tgggaggaag
      181 tttgccagga gtgatgaacg caagaggcat accaaaatcc atacaggtgc tagcttcatg
      241 aagatggaca aaaagactat agtttggttt agaagagatc taaggattga ggataatcct
      301 gcattagcag cagctgctca cgaaggatct gtttttcctg tcttcatttg gtgtcctgaa
      361 gaagaaggac agttttatcc tggaagagct tcaagatggt ggatgaaaca atcacttgct
      421 cacttatctc aatccttgaa ggctcttgga tctgacctca ctttaatcaa aacccacaac
      481 acgatttcag cgatcttgga ttgtatccgc gttaccggtg ctacaaaagt cgtctttaac
      541 cacctctatg atcctgtttc gttagttcgg gaccataccg taaaggagaa gctggtggaa
      601 cgtgggatct ctgtgcaaag ctacaatgga gatctattgt atgaaccgtg ggagatatac
      661 tgcgaaaagg gcaaaccttt tacgagtttc aattcttact ggaagaaatg cttagatatg
      721 tcgattgaat ccgttatgct tcctcctcct tggcggttga tgccaataac tgcagcggct
      781 gaagcgattt gggcgtgttc gattgaagaa ctagggctgg agaatgaggc cgagaaaccg
      841 agcaatgcgt tgttaactag agcttggagt ccaggatgga gcaatgctga taagttacta
      901 aatgagttca tcgagaagca gttgatagat tatgcaaaga acagcaagaa agttgttggg
      961 aattctactt cactactttc tccgtatctc catttcgggg aaataagcgt cagacacgtt
     1021 ttccagtgtg cccggatgaa acaaattata tgggcaagag ataagaacag tgaaggagaa
     1081 gaaagtgcag atctttttct taggggaatc ggtttaagag agtattctcg gtatatatgt
     1141 ttcaacttcc cgtttactca cgagcaatcg ttgttgagtc atcttcggtt tttcccttgg
     1201 gatgctgatg ttgataagtt caaggcctgg agacaaggca ggaccggtta tccgttggtg
     1261 gatgccggaa tgagagagct ttgggctacc ggatggatgc ataacagaat aagagtgatt
     1321 gtttcaagct ttgctgtgaa gtttcttctc cttccatgga aatggggaat gaagtatttc
     1381 tgggatacac ttttggatgc tgatttggaa tgtgacatcc ttggctggca gtatatctct
     1441 gggagtatcc ccgatggcca cgagcttgat cgcttggaca atcccgcgtt acaaggcgcc
     1501 aaatatgacc cagaaggtga gtacataagg caatggcttc ccgagcttgc gagattgcca
     1561 actgaatgga tccatcatcc atgggacgct cctttaaccg tactcaaagc ttctggtgtg
     1621 gaactcggaa caaactatgc gaaacccatt gtagacatcg acacagctcg tgagctacta
     1681 gctaaagcta tttcaagaac ccgtgaagca cagatcatga tcggagcagc acctgatgag
     1741 attgtagcag atagcttcga ggccttaggg gctaatacca ttaaagaacc tggtctttgc
     1801 ccatctgtgt cttctaatga ccaacaagta ccttcggctg ttcgttacaa cgggtcaaag
     1861 agagtgaaac ctgaggaaga agaagagaga gacatgaaga aatctagggg attcgatgaa
     1921 agggagttgt tttcgactgc tgaatcttct tcttcttcga gtgtgttttt cgtttcgcag
     1981 tcttgctcgt tggcatcaga agggaagaat ctggaaggta ttcaagattc atctgatcag
     2041 attactacaa gtttgggaaa aaatggttgc aaatgaggat ccgtcgacct gcagccaagc
     2101 taattccggg cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     2161 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     2221 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt
     2281 gaccacacct ctaccggcat ggagctccag cttttgttcc ctttagtgag ggttaattgc
     2341 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2401 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2461 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2521 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2581 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2641 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3781 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3961 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4261 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4381 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4441 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4501 agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta
     4561 atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt
     4621 tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt
     4681 gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca
     4741 cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta
     4801 atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta
     4861 aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc
     4921 taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc
     4981 atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtct aagaaaccat
     5041 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5101 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5161 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5221 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag
     5281 atcacgacat tactatatat ataatatagg aagcatttaa tagaacagca tcgtaatata
     5341 tgtgtacttt gcagttatga cgccagatgg cagtagtgga agatattctt tattgaaaaa
     5401 tagcttgtca ccttacgtac aatcttgatc cggagctttt ctttttttgc cgattaagaa
     5461 ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg agggcattgg tgactattga
     5521 gcacgtgagt atacgtgatt aagcacacaa aggcagcttg gagtatgtct gttattaatt
     5581 tcacaggtag ttctggtcca ttggtgaaag tttgcggctt gcagagcaca gaggccgcag
     5641 aatgtgctct agattccgat gctgacttgc tgggtattat atgtgtgccc aatagaaaga
     5701 gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata
     5761 gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt
     5821 tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat gagtcgtggc
     5881 aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact
     5941 gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt
     6001 cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa
     6061 ggcaagagag ccccgaaagc ttacatttta tgttagctgg tggactgacg ccagaaaatg
     6121 ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga
     6181 caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg
     6241 ttattactga gtagtattta tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg
     6301 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     6361 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     6421 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     6481 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6541 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6601 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6661 gggaaagccg gccaacttct tttctttttt tttcttttct ctctcccccg ttgttgtctc
     6721 accatatccg caatgacaaa aaaaatgatg gaagacacta aaggaaaaaa ttaacgacaa
     6781 agacagcacc aacagatgtc gttgttccag agctgatgag gggtatctcg aagcacacga
     6841 aactttttcc ttccttcatt cacgcacact actctctaat gagcaacggt atacggcctt
     6901 ccttccagtt acttgaattt gaaataaaaa aagtttgctg tcttgctatc aagtataaat
     6961 agacctgcaa ttattaatct tttgtttcct cgtcattgtt ctcgttccct ttcttccttg
     7021 tttctttttc tgcacaatat ttcaagctat accaagcata caatcaacta tctcatatac
     7081 aatggccccc aagaagaaga ggaaggttgg tattcacggg ggtacccgcc catatgcttg
     7141 ccctgtcgag tcc
//
