LOCUS       sequence-247341-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTEF1-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: pMM0620
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(21..125)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      228..816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     717..736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1395..1808
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1858..1878
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     2106..2125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2106..2124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2149..2658
                     /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(3712..4503,4504..4552,1..20))
                     /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     4335..4354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
       61 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      121 ccgcgcacat ttccccgaaa agtgccacct gtcatgacca aaatccctta acgtgagttt
      181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      361 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      841 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      901 tctgtggata accgtgcggc cgcccctgaa ttcgcatcta gatggtagag ccacaaacag
      961 ccggtacaag caacgatctc caggaccatc tgaatcatgc gcggatgaca cgaactcacg
     1021 acggcgatca cagacattaa cccacagtac agacactgcg acaacgtggc aattcgtcgc
     1081 aataccgtct cactgaactg gccgataatt gcagacgaac gccttgccaa cagggagttc
     1141 ttcagagaca tggaggctca aaacgaaatt attgacagcc tagacatcaa tagtcataca
     1201 acagaaagcg accacccaac tttggctgat aatagcgtat aaacaatgca tactttgtac
     1261 gttcaaaata caatgcagta gatatattta tgcatattac atataataca tatcacatag
     1321 gaagcaacag gcgcgttgga cttttaattt tcgaggaccg cgaatcctta catcacaccc
     1381 aatcccccac aagtgatccc ccacacacca tagcttcaaa atgtttctac tcctttttta
     1441 ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca
     1501 gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt
     1561 ttggaaaaga aaaaagacac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa
     1621 tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct ctttcgatga
     1681 cctcccattg atatttaagt taataaacgg tcatcaattt ctcaagtttc agtttcattt
     1741 ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct
     1801 aatctaagtt ttaattacaa aagatctatg gataaagcgg aattaattcc cgagcctcca
     1861 aaaaagaaga gaaaggtcga attgggtacc gccgcctcgg agctccactt agacggcgag
     1921 gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg atctggacat gttgggggac
     1981 ggggattccc cgggtccggg atttaccccc cacgactccg ccccctacgg cgctctggat
     2041 atggccgact tcgagtttga gcagatgttt accgatgccc ttggaattga cgagtacggt
     2101 gggtttaata cgactcacta tagggcgagc gccgaagcta gcgccaccat gaatggagct
     2161 ataggaggtg accttttgct caattttcct gacatgtcgg tcctagagcg ccaaagggct
     2221 cacctcaagt acctcaatcc cacctttgat tctcctctcg ccggcttctt tgccgattct
     2281 tcaatgatta ccggcggcga gatggacagc tatctttcga ctgccggttt gaatcttccg
     2341 atgatgtacg gtgaaacgac ggtggaaggt gattcaagac tctcaatttc gccggaaacg
     2401 acgcttggga ctggaaattt caagaaacgg aagtttgata cagagactaa ggattgtaat
     2461 gagaagaaga agaagatgac gatgaacaga gatgacctag tagaagaagg agaagaagag
     2521 aagtcgaaaa taacagagca aaacaatggg agcacaaaaa gcatcaagaa gatgaaacac
     2581 aaagccaaga aagaagagaa caatttctct aatgattcat ctaaagtgac gaaggaattg
     2641 gagaaaacgg attatattca tgttcgtgca cgacgaggcc aagccactga tagtcacagc
     2701 atagcagaac gagttagaag agaaaagatc agtgagagaa tgaagtttct acaagatttg
     2761 gttcctggat gcgacaagat cacaggcaaa gcagggatgc ttgatgaaat cattaactat
     2821 gttcagtctc ttcagagaca aatcgagttc ttatcgatga aactagcaat tgtgaatcca
     2881 aggccggatt ttgatatgga tgacattttt gccaaagagg ttgcctcaac tccaatgact
     2941 gtggtgccat ctcctgaaat ggttctttcc ggttattctc atgagatggt tcactctggt
     3001 tattctagtg agatggttaa ctccggttac cttcatgtca atccaatgca gcaagtgaat
     3061 accagttctg atccattgtc atgcttcaac aatggcgaag ctccttcgat gtgggactct
     3121 catgtgcaga atctctatgg caatttagga gtaccggtca tcgagctcga gctgcagatg
     3181 aatcgtagat acggatccta actcgagagc ttttgattaa gccttctagt ccaaaaaaca
     3241 cgtttttttg tcatttattt cattttctta gaatagttta gtttattcat tttatagtca
     3301 cgaatgtttt atgattctat atagggttgc aaacaagcat ttttcatttt atgttaaaac
     3361 aatttcaggt ttacctttta ttctgcttgt ggtgacgcgt gtatccgccc gctcttttgg
     3421 tcacccatgt atgctgccaa tgagacgacg gggtcatcac ggctcatcat gcgccaaaca
     3481 aatgtgtgca atacacgctc ggatgactgc atgatgaccg cactgactgg ggacagcaga
     3541 tccacctaag cctgtgagag aagcagacac ccgacagatc aaggcagtta actagtgcac
     3601 tgcagtacag cggccgcgat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3841 aatgataccg cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga ca
//
