LOCUS       sequence-258773-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRPL18B-VP16-CIB1 cassette plasmid flanked by ConLS and ConR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4552)
  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 4552)
  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: pMM0621
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             996..1016
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1244..1263
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1244..1262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1287..1796
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             complement(join(2850..3641,3642..3710))
                     /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     3473..3492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3711..3815)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3918..4506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4407..4426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
       61 cccctgaatt cgcatctaga tggtagagcc acaaacagcc ggtacaagca acgatctcca
      121 ggaccatctg aatcatgcgc ggatgacacg aactcacgac ggcgatcaca gacattaacc
      181 cacagtacag acactgcgac aacgtggcaa ttcgtcgcaa taccgtctca ctgaactggc
      241 cgataattgc agacgaacga agaggatgtc caatattttt tttaaggaat aaggatactt
      301 caagactaga ttcccccctg cattcccatc agaaccgtaa accttggcgc tttccttggg
      361 aagtattcaa gaagtgcctt gtccggtttc tgtggctcac aaaccagcgc gcccgatatg
      421 gctttctttt cacttatgaa tgtaccagta cgggacaatt agaacgctcc tgtaacaatc
      481 tctttgcaaa tgtggggtta cattctaacc atgtcacact gctgacgaaa ttcaaagtaa
      541 aaaaaaatgg gaccacgtct tgagaacgat agattttctt tattttacat tgaacagtcg
      601 ttgtctcagc gcgctttatg ttttcattca tacttcatat tataaaataa caaaagaaga
      661 atttcatatt cacgcccaag aaatcaggct gctttccaaa tgcaattgac acttcattag
      721 ccatcacaca aaactctttc ttgctggagc ttcttttaaa aaagacctca gtacaccaaa
      781 cacgttaccc gacctcgtta ttttacgaca actatgataa aattctgaag aaaaaataaa
      841 aaaattttca tacttcttgc ttttatttaa accattgaat gatttctttt gaacaaaact
      901 acctgtttca ccaaaggaaa tagaaagaaa aaatcaatta gaagaaaaca aaaaacaaaa
      961 gatctatgga taaagcggaa ttaattcccg agcctccaaa aaagaagaga aaggtcgaat
     1021 tgggtaccgc cgcctcggag ctccacttag acggcgagga cgtggcgatg gcgcatgccg
     1081 acgcgctaga cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat
     1141 ttacccccca cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc
     1201 agatgtttac cgatgccctt ggaattgacg agtacggtgg gtttaatacg actcactata
     1261 gggcgagcgc cgaagctagc gccaccatga atggagctat aggaggtgac cttttgctca
     1321 attttcctga catgtcggtc ctagagcgcc aaagggctca cctcaagtac ctcaatccca
     1381 cctttgattc tcctctcgcc ggcttctttg ccgattcttc aatgattacc ggcggcgaga
     1441 tggacagcta tctttcgact gccggtttga atcttccgat gatgtacggt gaaacgacgg
     1501 tggaaggtga ttcaagactc tcaatttcgc cggaaacgac gcttgggact ggaaatttca
     1561 agaaacggaa gtttgataca gagactaagg attgtaatga gaagaagaag aagatgacga
     1621 tgaacagaga tgacctagta gaagaaggag aagaagagaa gtcgaaaata acagagcaaa
     1681 acaatgggag cacaaaaagc atcaagaaga tgaaacacaa agccaagaaa gaagagaaca
     1741 atttctctaa tgattcatct aaagtgacga aggaattgga gaaaacggat tatattcatg
     1801 ttcgtgcacg acgaggccaa gccactgata gtcacagcat agcagaacga gttagaagag
     1861 aaaagatcag tgagagaatg aagtttctac aagatttggt tcctggatgc gacaagatca
     1921 caggcaaagc agggatgctt gatgaaatca ttaactatgt tcagtctctt cagagacaaa
     1981 tcgagttctt atcgatgaaa ctagcaattg tgaatccaag gccggatttt gatatggatg
     2041 acatttttgc caaagaggtt gcctcaactc caatgactgt ggtgccatct cctgaaatgg
     2101 ttctttccgg ttattctcat gagatggttc actctggtta ttctagtgag atggttaact
     2161 ccggttacct tcatgtcaat ccaatgcagc aagtgaatac cagttctgat ccattgtcat
     2221 gcttcaacaa tggcgaagct ccttcgatgt gggactctca tgtgcagaat ctctatggca
     2281 atttaggagt accggtcatc gagctcgagc tgcagatgaa tcgtagatac ggatcctaac
     2341 tcgagagctt ttgattaagc cttctagtcc aaaaaacacg tttttttgtc atttatttca
     2401 ttttcttaga atagtttagt ttattcattt tatagtcacg aatgttttat gattctatat
     2461 agggttgcaa acaagcattt ttcattttat gttaaaacaa tttcaggttt accttttatt
     2521 ctgcttgtgg tgacgcgtgt atccgcccgc tcttttggtc acccatgtat gctgccaatg
     2581 agacgacggg gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg
     2641 atgactgcat gatgaccgca ctgactgggg acagcagatc cacctaagcc tgtgagagaa
     2701 gcagacaccc gacagatcaa ggcagttaac tagtgcactg cagtacagcg gccgcgatta
     2761 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2821 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2881 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2941 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
     3001 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3061 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3121 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3181 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3241 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3301 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3361 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3421 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3481 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3541 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3601 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3661 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3721 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3781 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3841 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3901 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3961 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4021 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4081 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4141 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4201 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4261 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4321 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4381 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4441 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4501 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gc
//
