LOCUS       sequence-259406-        5332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTEF1-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: pMM0656
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             309..329
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             612..2447
                     /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(2967..3758,3759..3827))
                     /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     3590..3609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3828..3932)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4035..4623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4524..4543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5202..5332,1..283)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa gacaccgcct
       61 cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc tttttcttga
      121 aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata
      181 aacggtcatc aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt
      241 acttcttgct cattagaaag aaagcatagc aatctaatct aagttttaat tacaaaagat
      301 ctatggcccc caagaagaag aggaaggttg gtattcacgg gggtacccgc ccatatgctt
      361 gccctgtcga gtcctgcgat cgccgctttt ctcgctcgga tgagcttacc cgccatatcc
      421 gcatccatac cggtcagaag cccttccagt gtcgaatctg catgcgtaac ttcagtcgta
      481 gtgaccacct taccacccac atccgcaccc acacaggcga gaagcctttt gcctgtgaca
      541 tttgtgggag gaagtttgcc aggagtgatg aacgcaagag gcataccaaa atccatacag
      601 gtgctagctt catgaagatg gacaaaaaga ctatagtttg gtttagaaga gatctaagga
      661 ttgaggataa tcctgcatta gcagcagctg ctcacgaagg atctgttttt cctgtcttca
      721 tttggtgtcc tgaagaagaa ggacagtttt atcctggaag agcttcaaga tggtggatga
      781 aacaatcact tgctcactta tctcaatcct tgaaggctct tggatctgac ctcactttaa
      841 tcaaaaccca caacacgatt tcagcgatct tggattgtat ccgcgttacc ggtgctacaa
      901 aagtcgtctt taaccacctc tatgatcctg tttcgttagt tcgggaccat accgtaaagg
      961 agaagctggt ggaacgtggg atctctgtgc aaagctacaa tggagatcta ttgtatgaac
     1021 cgtgggagat atactgcgaa aagggcaaac cttttacgag tttcaattct tactggaaga
     1081 aatgcttaga tatgtcgatt gaatccgtta tgcttcctcc tccttggcgg ttgatgccaa
     1141 taactgcagc ggctgaagcg atttgggcgt gttcgattga agaactaggg ctggagaatg
     1201 aggccgagaa accgagcaat gcgttgttaa ctagagcttg gagtccagga tggagcaatg
     1261 ctgataagtt actaaatgag ttcatcgaga agcagttgat agattatgca aagaacagca
     1321 agaaagttgt tgggaattct acttcactac tttctccgta tctccatttc ggggaaataa
     1381 gcgtcagaca cgttttccag tgtgcccgga tgaaacaaat tatatgggca agagataaga
     1441 acagtgaagg agaagaaagt gcagatcttt ttcttagggg aatcggttta agagagtatt
     1501 ctcggtatat atgtttcaac ttcccgttta ctcacgagca atcgttgttg agtcatcttc
     1561 ggtttttccc ttgggatgct gatgttgata agttcaaggc ctggagacaa ggcaggaccg
     1621 gttatccgtt ggtggatgcc ggaatgagag agctttgggc taccggatgg atgcataaca
     1681 gaataagagt gattgtttca agctttgctg tgaagtttct tctccttcca tggaaatggg
     1741 gaatgaagta tttctgggat acacttttgg atgctgattt ggaatgtgac atccttggct
     1801 ggcagtatat ctctgggagt atccccgatg gccacgagct tgatcgcttg gacaatcccg
     1861 cgttacaagg cgccaaatat gacccagaag gtgagtacat aaggcaatgg cttcccgagc
     1921 ttgcgagatt gccaactgaa tggatccatc atccatggga cgctccttta accgtactca
     1981 aagcttctgg tgtggaactc ggaacaaact atgcgaaacc cattgtagac atcgacacag
     2041 ctcgtgagct actagctaaa gctatttcaa gaacccgtga agcacagatc atgatcggag
     2101 cagcacctga tgagattgta gcagatagct tcgaggcctt aggggctaat accattaaag
     2161 aacctggtct ttgcccatct gtgtcttcta atgaccaaca agtaccttcg gctgttcgtt
     2221 acaacgggtc aaagagagtg aaacctgagg aagaagaaga gagagacatg aagaaatcta
     2281 ggggattcga tgaaagggag ttgttttcga ctgctgaatc ttcttcttct tcgagtgtgt
     2341 ttttcgtttc gcagtcttgc tcgttggcat cagaagggaa gaatctggaa ggtattcaag
     2401 attcatctga tcagattact acaagtttgg gaaaaaatgg ttgcaaagga tcctaactcg
     2461 aggccaattg gtgcggcaat tgataataac gaaaatgtct tttaatgatc tgggtataat
     2521 gaggaatttt ccgaacgttt ttactttata tatatatata catgtaacat atattctata
     2581 cgctatagag aaaggaaatt tttcaattaa aaaaaaaata gagaaagagt ttcacttctt
     2641 gattatcgct aacactaatg gttgaagtac tgctacttta attttatgct ggatgtgaga
     2701 cgataaatcg cagccaagtg agtgaataga tgacgcacca cggtcagaca cggccacatc
     2761 gtatctcaca ggagcaagcg cgataggagc actcacacat agtacggtga tccgctgact
     2821 cctttgccca aataagacgt gagccactag tgcactgcag tacagcggcc gcgattatca
     2881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca
     3121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgtc
     3961 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4021 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4081 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4141 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4201 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4261 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4321 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4381 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4441 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4501 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4561 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4621 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4681 atgttctttc ctgcgttatc ccctgattct gtggataacc gtgcggccgc ccctgaattc
     4741 gcatctagaa cggggtcatc acggctcatc atgcgccaaa caaatgtgtg caatacacgc
     4801 tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta agcctgtgag
     4861 agaagcagac acccgacaga tcaaggcagt tacgtctcac caaaccagat gtcaacacag
     4921 ctacaacgcc ttgccaacag ggagttcttc agagacatgg aggctcaaaa cgaaattatt
     4981 gacagcctag acatcaatag tcatacaaca gaaagcgacc acccaacttt ggctgataat
     5041 agcgtataaa caatgcatac tttgtacgtt caaaatacaa tgcagtagat atatttatgc
     5101 atattacata taatacatat cacataggaa gcaacaggcg cgttggactt ttaattttcg
     5161 aggaccgcga atccttacat cacacccaat cccccacaag tgatccccca cacaccatag
     5221 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
     5281 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc tt
//
