LOCUS       sequence-247330-        7146 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 7146)
  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 7146)
  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..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /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(1378..1399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1572..1968
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1987..2007
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2290..4128
                     /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      4162..4349
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4217..4241)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4372..4392)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4372..4390)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4411..4427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4411..4427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4424..4446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4435..4451
                     /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(4459..4465,4466..4483,4484..4489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4642..4659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4813..5401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5572..6363,6364..6432))
                     /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     6195..6214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6433..6537)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6574..7077
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7119..7137
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg ccaacttctt ttcttttttt ttcttttctc tctcccccgt tgttgtctca
     1621 ccatatccgc aatgacaaaa aaaatgatgg aagacactaa aggaaaaaat taacgacaaa
     1681 gacagcacca acagatgtcg ttgttccaga gctgatgagg ggtatctcga agcacacgaa
     1741 actttttcct tccttcattc acgcacacta ctctctaatg agcaacggta tacggccttc
     1801 cttccagtta cttgaatttg aaataaaaaa agtttgctgt cttgctatca agtataaata
     1861 gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt tcttccttgt
     1921 ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaactat ctcatataca
     1981 atggccccca agaagaagag gaaggttggt attcacgggg gtacccgccc atatgcttgc
     2041 cctgtcgagt cctgcgatcg ccgcttttct cgctcggatg agcttacccg ccatatccgc
     2101 atccataccg gtcagaagcc cttccagtgt cgaatctgca tgcgtaactt cagtcgtagt
     2161 gaccacctta ccacccacat ccgcacccac acaggcgaga agccttttgc ctgtgacatt
     2221 tgtgggagga agtttgccag gagtgatgaa cgcaagaggc ataccaaaat ccatacaggt
     2281 gctagcttca tgaagatgga caaaaagact atagtttggt ttagaagaga tctaaggatt
     2341 gaggataatc ctgcattagc agcagctgct cacgaaggat ctgtttttcc tgtcttcatt
     2401 tggtgtcctg aagaagaagg acagttttat cctggaagag cttcaagatg gtggatgaaa
     2461 caatcacttg ctcacttatc tcaatccttg aaggctcttg gatctgacct cactttaatc
     2521 aaaacccaca acacgatttc agcgatcttg gattgtatcc gcgttaccgg tgctacaaaa
     2581 gtcgtcttta accacctcta tgatcctgtt tcgttagttc gggaccatac cgtaaaggag
     2641 aagctggtgg aacgtgggat ctctgtgcaa agctacaatg gagatctatt gtatgaaccg
     2701 tgggagatat actgcgaaaa gggcaaacct tttacgagtt tcaattctta ctggaagaaa
     2761 tgcttagata tgtcgattga atccgttatg cttcctcctc cttggcggtt gatgccaata
     2821 actgcagcgg ctgaagcgat ttgggcgtgt tcgattgaag aactagggct ggagaatgag
     2881 gccgagaaac cgagcaatgc gttgttaact agagcttgga gtccaggatg gagcaatgct
     2941 gataagttac taaatgagtt catcgagaag cagttgatag attatgcaaa gaacagcaag
     3001 aaagttgttg ggaattctac ttcactactt tctccgtatc tccatttcgg ggaaataagc
     3061 gtcagacacg ttttccagtg tgcccggatg aaacaaatta tatgggcaag agataagaac
     3121 agtgaaggag aagaaagtgc agatcttttt cttaggggaa tcggtttaag agagtattct
     3181 cggtatatat gtttcaactt cccgtttact cacgagcaat cgttgttgag tcatcttcgg
     3241 tttttccctt gggatgctga tgttgataag ttcaaggcct ggagacaagg caggaccggt
     3301 tatccgttgg tggatgccgg aatgagagag ctttgggcta ccggatggat gcataacaga
     3361 ataagagtga ttgtttcaag ctttgctgtg aagtttcttc tccttccatg gaaatgggga
     3421 atgaagtatt tctgggatac acttttggat gctgatttgg aatgtgacat ccttggctgg
     3481 cagtatatct ctgggagtat ccccgatggc cacgagcttg atcgcttgga caatcccgcg
     3541 ttacaaggcg ccaaatatga cccagaaggt gagtacataa ggcaatggct tcccgagctt
     3601 gcgagattgc caactgaatg gatccatcat ccatgggacg ctcctttaac cgtactcaaa
     3661 gcttctggtg tggaactcgg aacaaactat gcgaaaccca ttgtagacat cgacacagct
     3721 cgtgagctac tagctaaagc tatttcaaga acccgtgaag cacagatcat gatcggagca
     3781 gcacctgatg agattgtagc agatagcttc gaggccttag gggctaatac cattaaagaa
     3841 cctggtcttt gcccatctgt gtcttctaat gaccaacaag taccttcggc tgttcgttac
     3901 aacgggtcaa agagagtgaa acctgaggaa gaagaagaga gagacatgaa gaaatctagg
     3961 ggattcgatg aaagggagtt gttttcgact gctgaatctt cttcttcttc gagtgtgttt
     4021 ttcgtttcgc agtcttgctc gttggcatca gaagggaaga atctggaagg tattcaagat
     4081 tcatctgatc agattactac aagtttggga aaaaatggtt gcaaatgagg atccgtcgac
     4141 ctgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat tattaaataa
     4201 gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa
     4261 ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag
     4321 gtcgctctta ttgaccacac ctctaccggc atggagctcc agcttttgtt ccctttagtg
     4381 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4441 tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag cctggggtgc
     4501 ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4561 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4621 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5221 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6541 atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat
     6601 ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga
     6661 aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact
     6721 accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg
     6781 tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg
     6841 tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt
     6901 tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct
     6961 ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc
     7021 ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt
     7081 cctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7141 ttcgtc
//
