LOCUS       sequence-247364-        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"
     promoter        complement(17..121)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      224..812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     713..732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1830..1850
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2133..3968
                     /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(4488..5279,5280..5332,1..16))
                     /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     5111..5130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
       61 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      121 gcacatttcc ccgaaaagtg ccacctgtca tgaccaaaat cccttaacgt gagttttcgt
      181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      301 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      361 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      901 tggataaccg tgcggccgcc cctgaattcg catctagaac ggggtcatca cggctcatca
      961 tgcgccaaac aaatgtgtgc aatacacgct cggatgactg catgatgacc gcactgactg
     1021 gggacagcag atccacctaa gcctgtgaga gaagcagaca cccgacagat caaggcagtt
     1081 acgtctcacc aaaccagatg tcaacacagc tacaacgaag aggatgtcca atattttttt
     1141 taaggaataa ggatacttca agactagatt cccccctgca ttcccatcag aaccgtaaac
     1201 cttggcgctt tccttgggaa gtattcaaga agtgccttgt ccggtttctg tggctcacaa
     1261 accagcgcgc ccgatatggc tttcttttca cttatgaatg taccagtacg ggacaattag
     1321 aacgctcctg taacaatctc tttgcaaatg tggggttaca ttctaaccat gtcacactgc
     1381 tgacgaaatt caaagtaaaa aaaaatggga ccacgtcttg agaacgatag attttcttta
     1441 ttttacattg aacagtcgtt gtctcagcgc gctttatgtt ttcattcata cttcatatta
     1501 taaaataaca aaagaagaat ttcatattca cgcccaagaa atcaggctgc tttccaaatg
     1561 caattgacac ttcattagcc atcacacaaa actctttctt gctggagctt cttttaaaaa
     1621 agacctcagt acaccaaaca cgttacccga cctcgttatt ttacgacaac tatgataaaa
     1681 ttctgaagaa aaaataaaaa aattttcata cttcttgctt ttatttaaac cattgaatga
     1741 tttcttttga acaaaactac ctgtttcacc aaaggaaata gaaagaaaaa atcaattaga
     1801 agaaaacaaa aaacaaaaga tctatggccc ccaagaagaa gaggaaggtt ggtattcacg
     1861 ggggtacccg cccatatgct tgccctgtcg agtcctgcga tcgccgcttt tctcgctcgg
     1921 atgagcttac ccgccatatc cgcatccata ccggtcagaa gcccttccag tgtcgaatct
     1981 gcatgcgtaa cttcagtcgt agtgaccacc ttaccaccca catccgcacc cacacaggcg
     2041 agaagccttt tgcctgtgac atttgtggga ggaagtttgc caggagtgat gaacgcaaga
     2101 ggcataccaa aatccataca ggtgctagct tcatgaagat ggacaaaaag actatagttt
     2161 ggtttagaag agatctaagg attgaggata atcctgcatt agcagcagct gctcacgaag
     2221 gatctgtttt tcctgtcttc atttggtgtc ctgaagaaga aggacagttt tatcctggaa
     2281 gagcttcaag atggtggatg aaacaatcac ttgctcactt atctcaatcc ttgaaggctc
     2341 ttggatctga cctcacttta atcaaaaccc acaacacgat ttcagcgatc ttggattgta
     2401 tccgcgttac cggtgctaca aaagtcgtct ttaaccacct ctatgatcct gtttcgttag
     2461 ttcgggacca taccgtaaag gagaagctgg tggaacgtgg gatctctgtg caaagctaca
     2521 atggagatct attgtatgaa ccgtgggaga tatactgcga aaagggcaaa ccttttacga
     2581 gtttcaattc ttactggaag aaatgcttag atatgtcgat tgaatccgtt atgcttcctc
     2641 ctccttggcg gttgatgcca ataactgcag cggctgaagc gatttgggcg tgttcgattg
     2701 aagaactagg gctggagaat gaggccgaga aaccgagcaa tgcgttgtta actagagctt
     2761 ggagtccagg atggagcaat gctgataagt tactaaatga gttcatcgag aagcagttga
     2821 tagattatgc aaagaacagc aagaaagttg ttgggaattc tacttcacta ctttctccgt
     2881 atctccattt cggggaaata agcgtcagac acgttttcca gtgtgcccgg atgaaacaaa
     2941 ttatatgggc aagagataag aacagtgaag gagaagaaag tgcagatctt tttcttaggg
     3001 gaatcggttt aagagagtat tctcggtata tatgtttcaa cttcccgttt actcacgagc
     3061 aatcgttgtt gagtcatctt cggtttttcc cttgggatgc tgatgttgat aagttcaagg
     3121 cctggagaca aggcaggacc ggttatccgt tggtggatgc cggaatgaga gagctttggg
     3181 ctaccggatg gatgcataac agaataagag tgattgtttc aagctttgct gtgaagtttc
     3241 ttctccttcc atggaaatgg ggaatgaagt atttctggga tacacttttg gatgctgatt
     3301 tggaatgtga catccttggc tggcagtata tctctgggag tatccccgat ggccacgagc
     3361 ttgatcgctt ggacaatccc gcgttacaag gcgccaaata tgacccagaa ggtgagtaca
     3421 taaggcaatg gcttcccgag cttgcgagat tgccaactga atggatccat catccatggg
     3481 acgctccttt aaccgtactc aaagcttctg gtgtggaact cggaacaaac tatgcgaaac
     3541 ccattgtaga catcgacaca gctcgtgagc tactagctaa agctatttca agaacccgtg
     3601 aagcacagat catgatcgga gcagcacctg atgagattgt agcagatagc ttcgaggcct
     3661 taggggctaa taccattaaa gaacctggtc tttgcccatc tgtgtcttct aatgaccaac
     3721 aagtaccttc ggctgttcgt tacaacgggt caaagagagt gaaacctgag gaagaagaag
     3781 agagagacat gaagaaatct aggggattcg atgaaaggga gttgttttcg actgctgaat
     3841 cttcttcttc ttcgagtgtg tttttcgttt cgcagtcttg ctcgttggca tcagaaggga
     3901 agaatctgga aggtattcaa gattcatctg atcagattac tacaagtttg ggaaaaaatg
     3961 gttgcaaagg atcctaactc gaggccaatt ggtgcggcaa ttgataataa cgaaaatgtc
     4021 ttttaatgat ctgggtataa tgaggaattt tccgaacgtt tttactttat atatatatat
     4081 acatgtaaca tatattctat acgctataga gaaaggaaat ttttcaatta aaaaaaaaat
     4141 agagaaagag tttcacttct tgattatcgc taacactaat ggttgaagta ctgctacttt
     4201 aattttatgc tggatgtgag acgataaatc gcagccaagt gagtgaatag atgacgcacc
     4261 acggtcagac acggccacat cgtatctcac aggagcaagc gcgataggag cactcacaca
     4321 tagtacggtg atccgctgac tcctttgccc aaataagacg tgagccacta gtgcactgca
     4381 gtacagcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4441 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4501 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4561 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4621 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4681 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4741 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4801 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4861 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4921 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4981 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5041 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5101 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5161 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5221 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5281 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg ga
//
