LOCUS       sequence-247342-        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             741..761
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     989..1008
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        989..1007
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1032..1541
                     /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(2595..3386,3387..3455))
                     /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     3218..3237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3456..3560)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3663..4251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4152..4171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgaagagg atgtccaata ttttttttaa ggaataagga tacttcaaga ctagattccc
       61 ccctgcattc ccatcagaac cgtaaacctt ggcgctttcc ttgggaagta ttcaagaagt
      121 gccttgtccg gtttctgtgg ctcacaaacc agcgcgcccg atatggcttt cttttcactt
      181 atgaatgtac cagtacggga caattagaac gctcctgtaa caatctcttt gcaaatgtgg
      241 ggttacattc taaccatgtc acactgctga cgaaattcaa agtaaaaaaa aatgggacca
      301 cgtcttgaga acgatagatt ttctttattt tacattgaac agtcgttgtc tcagcgcgct
      361 ttatgttttc attcatactt catattataa aataacaaaa gaagaatttc atattcacgc
      421 ccaagaaatc aggctgcttt ccaaatgcaa ttgacacttc attagccatc acacaaaact
      481 ctttcttgct ggagcttctt ttaaaaaaga cctcagtaca ccaaacacgt tacccgacct
      541 cgttatttta cgacaactat gataaaattc tgaagaaaaa ataaaaaaat tttcatactt
      601 cttgctttta tttaaaccat tgaatgattt cttttgaaca aaactacctg tttcaccaaa
      661 ggaaatagaa agaaaaaatc aattagaaga aaacaaaaaa caaaagatct atggataaag
      721 cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt accgccgcct
      781 cggagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt
      841 tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact
      901 ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg
      961 cccttggaat tgacgagtac ggtgggttta atacgactca ctatagggcg agcgccgaag
     1021 ctagcgccac catgaatgga gctataggag gtgacctttt gctcaatttt cctgacatgt
     1081 cggtcctaga gcgccaaagg gctcacctca agtacctcaa tcccaccttt gattctcctc
     1141 tcgccggctt ctttgccgat tcttcaatga ttaccggcgg cgagatggac agctatcttt
     1201 cgactgccgg tttgaatctt ccgatgatgt acggtgaaac gacggtggaa ggtgattcaa
     1261 gactctcaat ttcgccggaa acgacgcttg ggactggaaa tttcaagaaa cggaagtttg
     1321 atacagagac taaggattgt aatgagaaga agaagaagat gacgatgaac agagatgacc
     1381 tagtagaaga aggagaagaa gagaagtcga aaataacaga gcaaaacaat gggagcacaa
     1441 aaagcatcaa gaagatgaaa cacaaagcca agaaagaaga gaacaatttc tctaatgatt
     1501 catctaaagt gacgaaggaa ttggagaaaa cggattatat tcatgttcgt gcacgacgag
     1561 gccaagccac tgatagtcac agcatagcag aacgagttag aagagaaaag atcagtgaga
     1621 gaatgaagtt tctacaagat ttggttcctg gatgcgacaa gatcacaggc aaagcaggga
     1681 tgcttgatga aatcattaac tatgttcagt ctcttcagag acaaatcgag ttcttatcga
     1741 tgaaactagc aattgtgaat ccaaggccgg attttgatat ggatgacatt tttgccaaag
     1801 aggttgcctc aactccaatg actgtggtgc catctcctga aatggttctt tccggttatt
     1861 ctcatgagat ggttcactct ggttattcta gtgagatggt taactccggt taccttcatg
     1921 tcaatccaat gcagcaagtg aataccagtt ctgatccatt gtcatgcttc aacaatggcg
     1981 aagctccttc gatgtgggac tctcatgtgc agaatctcta tggcaattta ggagtaccgg
     2041 tcatcgagct cgagctgcag atgaatcgta gatacggatc ctaactcgag agcttttgat
     2101 taagccttct agtccaaaaa acacgttttt ttgtcattta tttcattttc ttagaatagt
     2161 ttagtttatt cattttatag tcacgaatgt tttatgattc tatatagggt tgcaaacaag
     2221 catttttcat tttatgttaa aacaatttca ggtttacctt ttattctgct tgtggtgacg
     2281 cgtgtatccg cccgctcttt tggtcaccca tgtatgctgc caatgagacg acggggtcat
     2341 cacggctcat catgcgccaa acaaatgtgt gcaatacacg ctcggatgac tgcatgatga
     2401 ccgcactgac tggggacagc agatccacct aagcctgtga gagaagcaga cacccgacag
     2461 atcaaggcag ttaactagtg cactgcagta cagcggccgc gattatcaaa aaggatcttc
     2521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2641 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2701 ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat
     2761 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2821 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2881 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2941 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3001 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3061 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3121 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3181 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3241 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3301 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgtcat gaccaaaatc
     3601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3721 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3781 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3841 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3901 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3961 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4021 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4081 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4141 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4201 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4261 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4321 gcgttatccc ctgattctgt ggataaccgt gcggccgccc ctgaattcgc atctagatgg
     4381 tagagccaca aacagccggt acaagcaacg atctccagga ccatctgaat catgcgcgga
     4441 tgacacgaac tcacgacggc gatcacagac attaacccac agtacagaca ctgcgacaac
     4501 gtggcaattc gtcgcaatac cgtctcactg aactggccga taattgcaga cg
//
