LOCUS       sequence-258774-        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        411..824
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             874..894
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1122..1141
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1122..1140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1165..1674
                     /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(2728..3519,3520..3588))
                     /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     3351..3370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3589..3693)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3796..4384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4285..4304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accatctgaa tcatgcgcgg atgacacgaa ctcacgacgg cgatcacaga cattaaccca
       61 cagtacagac actgcgacaa cgtggcaatt cgtcgcaata ccgtctcact gaactggccg
      121 ataattgcag acgaacgcct tgccaacagg gagttcttca gagacatgga ggctcaaaac
      181 gaaattattg acagcctaga catcaatagt catacaacag aaagcgacca cccaactttg
      241 gctgataata gcgtataaac aatgcatact ttgtacgttc aaaatacaat gcagtagata
      301 tatttatgca tattacatat aatacatatc acataggaag caacaggcgc gttggacttt
      361 taattttcga ggaccgcgaa tccttacatc acacccaatc ccccacaagt gatcccccac
      421 acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc
      481 gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct
      541 tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agacaccgcc
      601 tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg
      661 aaaatttttt tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat
      721 aaacggtcat caatttctca agtttcagtt tcatttttct tgttctatta caactttttt
      781 tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttaa ttacaaaaga
      841 tctatggata aagcggaatt aattcccgag cctccaaaaa agaagagaaa ggtcgaattg
      901 ggtaccgccg cctcggagct ccacttagac ggcgaggacg tggcgatggc gcatgccgac
      961 gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg tccgggattt
     1021 accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga gtttgagcag
     1081 atgtttaccg atgcccttgg aattgacgag tacggtgggt ttaatacgac tcactatagg
     1141 gcgagcgccg aagctagcgc caccatgaat ggagctatag gaggtgacct tttgctcaat
     1201 tttcctgaca tgtcggtcct agagcgccaa agggctcacc tcaagtacct caatcccacc
     1261 tttgattctc ctctcgccgg cttctttgcc gattcttcaa tgattaccgg cggcgagatg
     1321 gacagctatc tttcgactgc cggtttgaat cttccgatga tgtacggtga aacgacggtg
     1381 gaaggtgatt caagactctc aatttcgccg gaaacgacgc ttgggactgg aaatttcaag
     1441 aaacggaagt ttgatacaga gactaaggat tgtaatgaga agaagaagaa gatgacgatg
     1501 aacagagatg acctagtaga agaaggagaa gaagagaagt cgaaaataac agagcaaaac
     1561 aatgggagca caaaaagcat caagaagatg aaacacaaag ccaagaaaga agagaacaat
     1621 ttctctaatg attcatctaa agtgacgaag gaattggaga aaacggatta tattcatgtt
     1681 cgtgcacgac gaggccaagc cactgatagt cacagcatag cagaacgagt tagaagagaa
     1741 aagatcagtg agagaatgaa gtttctacaa gatttggttc ctggatgcga caagatcaca
     1801 ggcaaagcag ggatgcttga tgaaatcatt aactatgttc agtctcttca gagacaaatc
     1861 gagttcttat cgatgaaact agcaattgtg aatccaaggc cggattttga tatggatgac
     1921 atttttgcca aagaggttgc ctcaactcca atgactgtgg tgccatctcc tgaaatggtt
     1981 ctttccggtt attctcatga gatggttcac tctggttatt ctagtgagat ggttaactcc
     2041 ggttaccttc atgtcaatcc aatgcagcaa gtgaatacca gttctgatcc attgtcatgc
     2101 ttcaacaatg gcgaagctcc ttcgatgtgg gactctcatg tgcagaatct ctatggcaat
     2161 ttaggagtac cggtcatcga gctcgagctg cagatgaatc gtagatacgg atcctaactc
     2221 gagagctttt gattaagcct tctagtccaa aaaacacgtt tttttgtcat ttatttcatt
     2281 ttcttagaat agtttagttt attcatttta tagtcacgaa tgttttatga ttctatatag
     2341 ggttgcaaac aagcattttt cattttatgt taaaacaatt tcaggtttac cttttattct
     2401 gcttgtggtg acgcgtgtat ccgcccgctc ttttggtcac ccatgtatgc tgccaatgag
     2461 acgacggggt catcacggct catcatgcgc caaacaaatg tgtgcaatac acgctcggat
     2521 gactgcatga tgaccgcact gactggggac agcagatcca cctaagcctg tgagagaagc
     2581 agacacccga cagatcaagg cagttaacta gtgcactgca gtacagcggc cgcgattatc
     2641 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2701 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2761 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2821 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     2881 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2941 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3001 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3061 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3121 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3181 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3241 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3301 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3361 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3421 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3481 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3541 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3601 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3661 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgt
     3721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3841 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4381 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4441 catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg cccctgaatt
     4501 cgcatctaga tggtagagcc acaaacagcc ggtacaagca acgatctcca gg
//
