LOCUS       sequence-247365-        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"
     promoter        complement(13..117)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      220..808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     709..728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1826..1846
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2129..3964
                     /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(4484..5275,5276..5332,1..12))
                     /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     5107..5126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
       61 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      121 atttccccga aaagtgccac ctgtcatgac caaaatccct taacgtgagt tttcgttcca
      181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      361 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      841 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      901 taaccgtgcg gccgcccctg aattcgcatc tagaacgggg tcatcacggc tcatcatgcg
      961 ccaaacaaat gtgtgcaata cacgctcgga tgactgcatg atgaccgcac tgactgggga
     1021 cagcagatcc acctaagcct gtgagagaag cagacacccg acagatcaag gcagttacgt
     1081 ctcaccaaac cagatgtcaa cacagctaca acgagtcgaa caagaagcag gcaaagttta
     1141 gagcactgcc cctccgcact caaaaaagaa aaaactagga ggaaaataaa attctcaacc
     1201 acacaaacac ataaacacat acaaatacaa atacaagctt atttacttga catcgcgcga
     1261 tcttccacta ttcagcgccg tccgccctct ctcgtgtttt ttgtttacgc gacaactatg
     1321 cgaaatccgg agcaacgggc aaccgtttgg ggaaagacca cacccacgcg cgatcgccat
     1381 ggcaacgagg tcgcacacgc cccacaccca gacctccctg cgagcgggca tgggtacaat
     1441 gtccccgttg ccacagacac cacttcgtag cacagcgcag agcgtagcgt gttgttgctg
     1501 ctgacaaaag aaaatttttc ttagcaaagc aaaggagggg aagcacgggc agatagcacc
     1561 gtaccatacc cttggaaact cgaaatgaac gaagcaggaa atgagagaat gagagttttg
     1621 taggtatata tagcggtagt gtttgcgcgt taccatcatc ttctggatct atctattgtt
     1681 cttttcctca tcactttccc ctttttcgct cttcttcttg tcttttattt ctttcttttt
     1741 tttaattgtt ccctcgattg gctatctacc aaagaatcca aacttaatac acgtatttat
     1801 ttgtccaatt accagatcta tggcccccaa gaagaagagg aaggttggta ttcacggggg
     1861 tacccgccca tatgcttgcc ctgtcgagtc ctgcgatcgc cgcttttctc gctcggatga
     1921 gcttacccgc catatccgca tccataccgg tcagaagccc ttccagtgtc gaatctgcat
     1981 gcgtaacttc agtcgtagtg accaccttac cacccacatc cgcacccaca caggcgagaa
     2041 gccttttgcc tgtgacattt gtgggaggaa gtttgccagg agtgatgaac gcaagaggca
     2101 taccaaaatc catacaggtg ctagcttcat gaagatggac aaaaagacta tagtttggtt
     2161 tagaagagat ctaaggattg aggataatcc tgcattagca gcagctgctc acgaaggatc
     2221 tgtttttcct gtcttcattt ggtgtcctga agaagaagga cagttttatc ctggaagagc
     2281 ttcaagatgg tggatgaaac aatcacttgc tcacttatct caatccttga aggctcttgg
     2341 atctgacctc actttaatca aaacccacaa cacgatttca gcgatcttgg attgtatccg
     2401 cgttaccggt gctacaaaag tcgtctttaa ccacctctat gatcctgttt cgttagttcg
     2461 ggaccatacc gtaaaggaga agctggtgga acgtgggatc tctgtgcaaa gctacaatgg
     2521 agatctattg tatgaaccgt gggagatata ctgcgaaaag ggcaaacctt ttacgagttt
     2581 caattcttac tggaagaaat gcttagatat gtcgattgaa tccgttatgc ttcctcctcc
     2641 ttggcggttg atgccaataa ctgcagcggc tgaagcgatt tgggcgtgtt cgattgaaga
     2701 actagggctg gagaatgagg ccgagaaacc gagcaatgcg ttgttaacta gagcttggag
     2761 tccaggatgg agcaatgctg ataagttact aaatgagttc atcgagaagc agttgataga
     2821 ttatgcaaag aacagcaaga aagttgttgg gaattctact tcactacttt ctccgtatct
     2881 ccatttcggg gaaataagcg tcagacacgt tttccagtgt gcccggatga aacaaattat
     2941 atgggcaaga gataagaaca gtgaaggaga agaaagtgca gatctttttc ttaggggaat
     3001 cggtttaaga gagtattctc ggtatatatg tttcaacttc ccgtttactc acgagcaatc
     3061 gttgttgagt catcttcggt ttttcccttg ggatgctgat gttgataagt tcaaggcctg
     3121 gagacaaggc aggaccggtt atccgttggt ggatgccgga atgagagagc tttgggctac
     3181 cggatggatg cataacagaa taagagtgat tgtttcaagc tttgctgtga agtttcttct
     3241 ccttccatgg aaatggggaa tgaagtattt ctgggataca cttttggatg ctgatttgga
     3301 atgtgacatc cttggctggc agtatatctc tgggagtatc cccgatggcc acgagcttga
     3361 tcgcttggac aatcccgcgt tacaaggcgc caaatatgac ccagaaggtg agtacataag
     3421 gcaatggctt cccgagcttg cgagattgcc aactgaatgg atccatcatc catgggacgc
     3481 tcctttaacc gtactcaaag cttctggtgt ggaactcgga acaaactatg cgaaacccat
     3541 tgtagacatc gacacagctc gtgagctact agctaaagct atttcaagaa cccgtgaagc
     3601 acagatcatg atcggagcag cacctgatga gattgtagca gatagcttcg aggccttagg
     3661 ggctaatacc attaaagaac ctggtctttg cccatctgtg tcttctaatg accaacaagt
     3721 accttcggct gttcgttaca acgggtcaaa gagagtgaaa cctgaggaag aagaagagag
     3781 agacatgaag aaatctaggg gattcgatga aagggagttg ttttcgactg ctgaatcttc
     3841 ttcttcttcg agtgtgtttt tcgtttcgca gtcttgctcg ttggcatcag aagggaagaa
     3901 tctggaaggt attcaagatt catctgatca gattactaca agtttgggaa aaaatggttg
     3961 caaaggatcc taactcgagg ccaattggtg cggcaattga taataacgaa aatgtctttt
     4021 aatgatctgg gtataatgag gaattttccg aacgttttta ctttatatat atatatacat
     4081 gtaacatata ttctatacgc tatagagaaa ggaaattttt caattaaaaa aaaaatagag
     4141 aaagagtttc acttcttgat tatcgctaac actaatggtt gaagtactgc tactttaatt
     4201 ttatgctgga tgtgagacga taaatcgcag ccaagtgagt gaatagatga cgcaccacgg
     4261 tcagacacgg ccacatcgta tctcacagga gcaagcgcga taggagcact cacacatagt
     4321 acggtgatcc gctgactcct ttgcccaaat aagacgtgag ccactagtgc actgcagtac
     4381 agcggccgcg attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4621 cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tg
//
