LOCUS       sequence-259411-        5332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRPL18B-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: pMM0657
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             205..2040
                     /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(2560..3351,3352..3420))
                     /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     3183..3202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3421..3525)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3628..4216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4117..4136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5234..5254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 acccgccata tccgcatcca taccggtcag aagcccttcc agtgtcgaat ctgcatgcgt
       61 aacttcagtc gtagtgacca ccttaccacc cacatccgca cccacacagg cgagaagcct
      121 tttgcctgtg acatttgtgg gaggaagttt gccaggagtg atgaacgcaa gaggcatacc
      181 aaaatccata caggtgctag cttcatgaag atggacaaaa agactatagt ttggtttaga
      241 agagatctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
      301 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
      361 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
      421 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
      481 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
      541 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
      601 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
      661 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
      721 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
      781 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggagtcca
      841 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
      901 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
      961 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     1021 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     1081 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     1141 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     1201 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     1261 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     1321 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     1381 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     1441 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     1501 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     1561 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     1621 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
     1681 atcatgatcg gagcagcacc tgatgagatt gtagcagata gcttcgaggc cttaggggct
     1741 aataccatta aagaacctgg tctttgccca tctgtgtctt ctaatgacca acaagtacct
     1801 tcggctgttc gttacaacgg gtcaaagaga gtgaaacctg aggaagaaga agagagagac
     1861 atgaagaaat ctaggggatt cgatgaaagg gagttgtttt cgactgctga atcttcttct
     1921 tcttcgagtg tgtttttcgt ttcgcagtct tgctcgttgg catcagaagg gaagaatctg
     1981 gaaggtattc aagattcatc tgatcagatt actacaagtt tgggaaaaaa tggttgcaaa
     2041 ggatcctaac tcgaggccaa ttggtgcggc aattgataat aacgaaaatg tcttttaatg
     2101 atctgggtat aatgaggaat tttccgaacg tttttacttt atatatatat atacatgtaa
     2161 catatattct atacgctata gagaaaggaa atttttcaat taaaaaaaaa atagagaaag
     2221 agtttcactt cttgattatc gctaacacta atggttgaag tactgctact ttaattttat
     2281 gctggatgtg agacgataaa tcgcagccaa gtgagtgaat agatgacgca ccacggtcag
     2341 acacggccac atcgtatctc acaggagcaa gcgcgatagg agcactcaca catagtacgg
     2401 tgatccgctg actcctttgc ccaaataaga cgtgagccac tagtgcactg cagtacagcg
     2461 gccgcgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2701 ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3541 agtgccacct gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc
     4321 cgcccctgaa ttcgcatcta gaacggggtc atcacggctc atcatgcgcc aaacaaatgt
     4381 gtgcaataca cgctcggatg actgcatgat gaccgcactg actggggaca gcagatccac
     4441 ctaagcctgt gagagaagca gacacccgac agatcaaggc agttacgtct caccaaacca
     4501 gatgtcaaca cagctacaac gaagaggatg tccaatattt tttttaagga ataaggatac
     4561 ttcaagacta gattcccccc tgcattccca tcagaaccgt aaaccttggc gctttccttg
     4621 ggaagtattc aagaagtgcc ttgtccggtt tctgtggctc acaaaccagc gcgcccgata
     4681 tggctttctt ttcacttatg aatgtaccag tacgggacaa ttagaacgct cctgtaacaa
     4741 tctctttgca aatgtggggt tacattctaa ccatgtcaca ctgctgacga aattcaaagt
     4801 aaaaaaaaat gggaccacgt cttgagaacg atagattttc tttattttac attgaacagt
     4861 cgttgtctca gcgcgcttta tgttttcatt catacttcat attataaaat aacaaaagaa
     4921 gaatttcata ttcacgccca agaaatcagg ctgctttcca aatgcaattg acacttcatt
     4981 agccatcaca caaaactctt tcttgctgga gcttctttta aaaaagacct cagtacacca
     5041 aacacgttac ccgacctcgt tattttacga caactatgat aaaattctga agaaaaaata
     5101 aaaaaatttt catacttctt gcttttattt aaaccattga atgatttctt ttgaacaaaa
     5161 ctacctgttt caccaaagga aatagaaaga aaaaatcaat tagaagaaaa caaaaaacaa
     5221 aagatctatg gcccccaaga agaagaggaa ggttggtatt cacgggggta cccgcccata
     5281 tgcttgccct gtcgagtcct gcgatcgccg cttttctcgc tcggatgagc tt
//
