LOCUS       sequence-259400-        7153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Zif268DBD-CRY2 under pscADH1.
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: pMM0317
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             207..2045
                     /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      2079..2266
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2134..2158)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2289..2309)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2289..2307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2328..2344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2328..2344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2341..2363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2352..2368
                     /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(2376..2382,2383..2400,2401..2406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2421..2442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2559..2576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2730..3318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2810..2829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3489..4280,4281..4349))
                     /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     4112..4131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4350..4454)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4491..4994
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5035..5053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5091..5113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5213..5232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5253..5533
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5534..6208
                     /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(6448..6469)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6642..7038
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             7057..7077
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ttacccgcca tatccgcatc cataccggtc agaagccctt ccagtgtcga atctgcatgc
       61 gtaacttcag tcgtagtgac caccttacca cccacatccg cacccacaca ggcgagaagc
      121 cttttgcctg tgacatttgt gggaggaagt ttgccaggag tgatgaacgc aagaggcata
      181 ccaaaatcca tacaggtgct agcttcatga agatggacaa aaagactata gtttggttta
      241 gaagagacct aaggattgag gataatcctg cattagcagc agctgctcac gaaggatctg
      301 tttttcctgt cttcatttgg tgtcctgaag aagaaggaca gttttatcct ggaagagctt
      361 caagatggtg gatgaaacaa tcacttgctc acttatctca atccttgaag gctcttggat
      421 ctgacctcac tttaatcaaa acccacaaca cgatttcagc gatcttggat tgtatccgcg
      481 ttaccggtgc tacaaaagtc gtctttaacc acctctatga tcctgtttcg ttagttcggg
      541 accataccgt aaaggagaag ctggtggaac gtgggatctc tgtgcaaagc tacaatggag
      601 atctattgta tgaaccgtgg gagatatact gcgaaaaggg caaacctttt acgagtttca
      661 attcttactg gaagaaatgc ttagatatgt cgattgaatc cgttatgctt cctcctcctt
      721 ggcggttgat gccaataact gcagcggctg aagcgatttg ggcgtgttcg attgaagaac
      781 tagggctgga gaatgaggcc gagaaaccga gcaatgcgtt gttaactaga gcttggtctc
      841 caggatggag caatgctgat aagttactaa atgagttcat cgagaagcag ttgatagatt
      901 atgcaaagaa cagcaagaaa gttgttggga attctacttc actactttct ccgtatctcc
      961 atttcgggga aataagcgtc agacacgttt tccagtgtgc ccggatgaaa caaattatat
     1021 gggcaagaga taagaacagt gaaggagaag aaagtgcaga tctttttctt aggggaatcg
     1081 gtttaagaga gtattctcgg tatatatgtt tcaacttccc gtttactcac gagcaatcgt
     1141 tgttgagtca tcttcggttt ttcccttggg atgctgatgt tgataagttc aaggcctgga
     1201 gacaaggcag gaccggttat ccgttggtgg atgccggaat gagagagctt tgggctaccg
     1261 gatggatgca taacagaata agagtgattg tttcaagctt tgctgtgaag tttcttctcc
     1321 ttccatggaa atggggaatg aagtatttct gggatacact tttggatgct gatttggaat
     1381 gtgacatcct tggctggcag tatatctctg ggagtatccc cgatggccac gagcttgatc
     1441 gcttggacaa tcccgcgtta caaggcgcca aatatgaccc agaaggtgag tacataaggc
     1501 aatggcttcc cgagcttgcg agattgccaa ctgaatggat ccatcatcca tgggacgctc
     1561 ctttaaccgt actcaaagct tctggtgtgg aactcggaac aaactatgcg aaacccattg
     1621 tagacatcga cacagctcgt gagctactag ctaaagctat ttcaagaacc cgtgaagcac
     1681 agatcatgat cggagcagca cctgatgaga ttgtagcaga tagcttcgag gccttagggg
     1741 ctaataccat taaagaacct ggtctttgcc catctgtgtc ttctaatgac caacaagtac
     1801 cttcggctgt tcgttacaac gggtcaaaga gagtgaaacc tgaggaagaa gaagagagag
     1861 acatgaagaa atctagggga ttcgatgaaa gggagttgtt ttcgactgct gaatcttctt
     1921 cttcttcgag tgtgtttttc gtttcgcagt cttgctcgtt ggcatcagaa gggaagaatc
     1981 tggaaggtat tcaagattca tctgatcaga ttactacaag tttgggaaaa aatggttgca
     2041 aatgaggatc cgtcgacctg cagccaagct aattccgggc gaatttctta tgatttatga
     2101 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     2161 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     2221 ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg gagctccagc
     2281 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2341 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2401 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2461 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2521 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2581 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2641 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2701 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2761 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2821 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2881 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2941 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3001 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3061 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3121 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3181 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3241 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3301 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3361 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3421 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3481 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3721 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3781 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3901 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4021 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4141 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4201 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4321 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4381 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4441 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc atcacgtgct
     4501 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     4561 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     4621 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     4681 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac
     4741 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     4801 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     4861 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     4921 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     4981 acaggcaagc gatccgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5041 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5101 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5161 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     5221 gagcagattg tactgagagt gcaccataga tcacgacatt actatatata taatatagga
     5281 agcatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac gccagatggc
     5341 agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca atcttgatcc
     5401 ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa aaggagaggg
     5461 ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa
     5521 ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt
     5581 ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg ctgacttgct
     5641 gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat
     5701 ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt
     5761 gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat
     5821 cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg gtttgccagt
     5881 tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa
     5941 acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg aacttttgga
     6001 ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct tacattttat
     6061 gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa atggcgttat
     6121 tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta acaaaatagc
     6181 aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat ttaagtattg
     6241 tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6301 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6361 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6421 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6481 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6541 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6601 aaagggagcc cccgatttag agcttgacgg ggaaagccgg ccaacttctt ttcttttttt
     6661 ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa aaaatgatgg
     6721 aagacactaa aggaaaaaat taacgacaaa gacagcacca acagatgtcg ttgttccaga
     6781 gctgatgagg ggtatctcga agcacacgaa actttttcct tccttcattc acgcacacta
     6841 ctctctaatg agcaacggta tacggccttc cttccagtta cttgaatttg aaataaaaaa
     6901 agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc
     6961 gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata
     7021 ccaagcatac aatcaactat ctcatataca atggccccca agaagaagag gaaggttggt
     7081 attcacgggg gtacccgccc atatgcttgc cctgtcgagt cctgcgatcg ccgcttttct
     7141 cgctcggatg agc
//
