LOCUS       sequence-259417-        5332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pREV1-Zif268DBD-CRY2 cassette plasmid flanked by ConL1 and ConR2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5332)
  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 5332)
  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: pMM0658
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             201..2036
                     /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"
     CDS             complement(join(2556..3347,3348..3416))
                     /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     3179..3198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3417..3521)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3624..4212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4113..4132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5230..5250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gccatatccg catccatacc ggtcagaagc ccttccagtg tcgaatctgc atgcgtaact
       61 tcagtcgtag tgaccacctt accacccaca tccgcaccca cacaggcgag aagccttttg
      121 cctgtgacat ttgtgggagg aagtttgcca ggagtgatga acgcaagagg cataccaaaa
      181 tccatacagg tgctagcttc atgaagatgg acaaaaagac tatagtttgg tttagaagag
      241 atctaaggat tgaggataat cctgcattag cagcagctgc tcacgaagga tctgtttttc
      301 ctgtcttcat ttggtgtcct gaagaagaag gacagtttta tcctggaaga gcttcaagat
      361 ggtggatgaa acaatcactt gctcacttat ctcaatcctt gaaggctctt ggatctgacc
      421 tcactttaat caaaacccac aacacgattt cagcgatctt ggattgtatc cgcgttaccg
      481 gtgctacaaa agtcgtcttt aaccacctct atgatcctgt ttcgttagtt cgggaccata
      541 ccgtaaagga gaagctggtg gaacgtggga tctctgtgca aagctacaat ggagatctat
      601 tgtatgaacc gtgggagata tactgcgaaa agggcaaacc ttttacgagt ttcaattctt
      661 actggaagaa atgcttagat atgtcgattg aatccgttat gcttcctcct ccttggcggt
      721 tgatgccaat aactgcagcg gctgaagcga tttgggcgtg ttcgattgaa gaactagggc
      781 tggagaatga ggccgagaaa ccgagcaatg cgttgttaac tagagcttgg agtccaggat
      841 ggagcaatgc tgataagtta ctaaatgagt tcatcgagaa gcagttgata gattatgcaa
      901 agaacagcaa gaaagttgtt gggaattcta cttcactact ttctccgtat ctccatttcg
      961 gggaaataag cgtcagacac gttttccagt gtgcccggat gaaacaaatt atatgggcaa
     1021 gagataagaa cagtgaagga gaagaaagtg cagatctttt tcttagggga atcggtttaa
     1081 gagagtattc tcggtatata tgtttcaact tcccgtttac tcacgagcaa tcgttgttga
     1141 gtcatcttcg gtttttccct tgggatgctg atgttgataa gttcaaggcc tggagacaag
     1201 gcaggaccgg ttatccgttg gtggatgccg gaatgagaga gctttgggct accggatgga
     1261 tgcataacag aataagagtg attgtttcaa gctttgctgt gaagtttctt ctccttccat
     1321 ggaaatgggg aatgaagtat ttctgggata cacttttgga tgctgatttg gaatgtgaca
     1381 tccttggctg gcagtatatc tctgggagta tccccgatgg ccacgagctt gatcgcttgg
     1441 acaatcccgc gttacaaggc gccaaatatg acccagaagg tgagtacata aggcaatggc
     1501 ttcccgagct tgcgagattg ccaactgaat ggatccatca tccatgggac gctcctttaa
     1561 ccgtactcaa agcttctggt gtggaactcg gaacaaacta tgcgaaaccc attgtagaca
     1621 tcgacacagc tcgtgagcta ctagctaaag ctatttcaag aacccgtgaa gcacagatca
     1681 tgatcggagc agcacctgat gagattgtag cagatagctt cgaggcctta ggggctaata
     1741 ccattaaaga acctggtctt tgcccatctg tgtcttctaa tgaccaacaa gtaccttcgg
     1801 ctgttcgtta caacgggtca aagagagtga aacctgagga agaagaagag agagacatga
     1861 agaaatctag gggattcgat gaaagggagt tgttttcgac tgctgaatct tcttcttctt
     1921 cgagtgtgtt tttcgtttcg cagtcttgct cgttggcatc agaagggaag aatctggaag
     1981 gtattcaaga ttcatctgat cagattacta caagtttggg aaaaaatggt tgcaaaggat
     2041 cctaactcga ggccaattgg tgcggcaatt gataataacg aaaatgtctt ttaatgatct
     2101 gggtataatg aggaattttc cgaacgtttt tactttatat atatatatac atgtaacata
     2161 tattctatac gctatagaga aaggaaattt ttcaattaaa aaaaaaatag agaaagagtt
     2221 tcacttcttg attatcgcta acactaatgg ttgaagtact gctactttaa ttttatgctg
     2281 gatgtgagac gataaatcgc agccaagtga gtgaatagat gacgcaccac ggtcagacac
     2341 ggccacatcg tatctcacag gagcaagcgc gataggagca ctcacacata gtacggtgat
     2401 ccgctgactc ctttgcccaa ataagacgtg agccactagt gcactgcagt acagcggccg
     2461 cgattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2521 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2581 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2641 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgggac
     2701 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2761 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2821 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2881 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2941 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3001 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3061 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3121 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3181 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3241 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3301 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3361 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3421 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3481 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3541 ccacctgtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4081 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4141 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4201 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4261 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tgcggccgcc
     4321 cctgaattcg catctagaac ggggtcatca cggctcatca tgcgccaaac aaatgtgtgc
     4381 aatacacgct cggatgactg catgatgacc gcactgactg gggacagcag atccacctaa
     4441 gcctgtgaga gaagcagaca cccgacagat caaggcagtt acgtctcacc aaaccagatg
     4501 tcaacacagc tacaacgagt cgaacaagaa gcaggcaaag tttagagcac tgcccctccg
     4561 cactcaaaaa agaaaaaact aggaggaaaa taaaattctc aaccacacaa acacataaac
     4621 acatacaaat acaaatacaa gcttatttac ttgacatcgc gcgatcttcc actattcagc
     4681 gccgtccgcc ctctctcgtg ttttttgttt acgcgacaac tatgcgaaat ccggagcaac
     4741 gggcaaccgt ttggggaaag accacaccca cgcgcgatcg ccatggcaac gaggtcgcac
     4801 acgccccaca cccagacctc cctgcgagcg ggcatgggta caatgtcccc gttgccacag
     4861 acaccacttc gtagcacagc gcagagcgta gcgtgttgtt gctgctgaca aaagaaaatt
     4921 tttcttagca aagcaaagga ggggaagcac gggcagatag caccgtacca tacccttgga
     4981 aactcgaaat gaacgaagca ggaaatgaga gaatgagagt tttgtaggta tatatagcgg
     5041 tagtgtttgc gcgttaccat catcttctgg atctatctat tgttcttttc ctcatcactt
     5101 tccccttttt cgctcttctt cttgtctttt atttctttct tttttttaat tgttccctcg
     5161 attggctatc taccaaagaa tccaaactta atacacgtat ttatttgtcc aattaccaga
     5221 tctatggccc ccaagaagaa gaggaaggtt ggtattcacg ggggtacccg cccatatgct
     5281 tgccctgtcg agtcctgcga tcgccgcttt tctcgctcgg atgagcttac cc
//
