LOCUS       sequence-314367-        4074 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Expresses LightR-bRaf-mVenus in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4074)
  AUTHORS   Shaaya M, Fauser J, Zhurikhina A, Conage-Pough JE, Huyot V, Brennan 
            M, Flower CT, Matsche J, Khan S, Natarajan V, Rehman J, Kota P, 
            White FM, Tsygankov D, Karginov AV
  TITLE     Light-Regulated allosteric switch enables temporal and subcellular 
            control of enzyme activity.
  JOURNAL   Elife. 2020 Sep 23;9. pii: 60647. doi: 10.7554/eLife.60647.
  PUBMED    32965214
REFERENCE   2  (bases 1 to 4074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2020-09-23"; volume: "9. pii: 60647. doi"; pages: " 
            10.7554/eLife.60647"
COMMENT     Sequence Label: LightR-bRaf-mVenus
FEATURES             Location/Qualifiers
     source          1..4074
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1966..2415
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLIQIMNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     regulatory      3352..3361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3358..3360,3361..3363,3364..4074)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3403..3424)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3664..3683)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4011..4032
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atggcggcgc tgagcggtgg cggtggtggc ggcgcggagc cgggccaggc tctgttcaac
       61 ggggacatgg agcccgaggc cggcgccggc gccggcgccg cggcctcttc ggctgcggac
      121 cctgccattc cggaggaggt gtggaatatc aaacaaatga ttaagttgac acaggaacat
      181 atagaggccc tattggacaa atttggtggg gagcataatc caccatcaat atatctggag
      241 gcctatgaag aatacaccag caagctagat gcactccaac aaagagaaca acagttattg
      301 gaatctctgg ggaacggaac tgatttttct gtttctagct ctgcatcaat ggataccgtt
      361 acatcttctt cctcttctag cctttcagtg ctaccttcat ctctttcagt ttttcaaaat
      421 cccacagatg tggcacggag caaccccaag tcaccacaaa aacctatcgt tagagtcttc
      481 ctgcccaaca aacagaggac agtggtacct gcaaggtgtg gagttacagt ccgagacagt
      541 ctaaagaaag cactgatgat gagaggtcta atcccagagt gctgtgctgt ttacagaatt
      601 caggatggag agaagaaacc aattggttgg gacactgata tttcctggct tactggagaa
      661 gaattgcatg tggaagtgtt ggagaatgtt ccacttacaa cacacaactt tgtacgaaaa
      721 acgtttttca ccttagcatt ttgtgacttt tgtcgaaagc tgcttttcca gggtttccgc
      781 tgtcaaacat gtggttataa atttcaccag cgttgtagta cagaagttcc actgatgtgt
      841 gttaattatg accaacttga tttgctgttt gtctccaagt tctttgaaca ccacccaata
      901 ccacaggaag aggcgtcctt agcagagact gccctaacat ctggatcatc cccttccgca
      961 cccgcctcgg actctattgg gccccaaatt ctcaccagtc cgtctccttc aaaatccatt
     1021 ccaattccac agcccttccg accagcagat gaagatcatc gaaatcaatt tgggcaacga
     1081 gaccgatcct catcagctcc caatgtgcat ataaacacaa tagaacctgt caatattgat
     1141 gacttgatta gagaccaagg atttcgtggt gatggaggat caaccacagg tttgtctgct
     1201 accccccctg cctcattacc tggctcacta actaacgtga aagccttaca gaaatctcca
     1261 ggacctcagc gagaaaggaa gtcatcttca tcctcagaag acaggaatcg aatgaaaaca
     1321 cttggtagac gggactcgag tgatgattgg gagattcctg atgggcagat tacagtggga
     1381 caaagaattg gatctggatc atttggaaca gtctacaagg gaaagtgggg accaggtggc
     1441 agcggaggtc ataccttgta tgcgccgggg ggttatgaca tcatgggtta cctcatacag
     1501 atcatgaata ggccgaaccc acaagtggag ctcggacccg tcgatacctc ctgcgctctc
     1561 attctgtgtg accttaagca gaaggatacc cctatcgtgt acgcctccga ggcatttctg
     1621 tacatgacag ggtactcgaa cgccgaagtg ctgggacgga actgccgctt cctgcaaagc
     1681 ccggatggaa tggtgaagcc taagtcaacc cgcaaatacg tggactccaa cactatcaac
     1741 accatgcgca aggccattga ccgcaatgct gaggtgcaag tggaagtggt gaacttcaag
     1801 aagaatggac agcgcttcgt caacttcctg actatgatcc ccgtgcgcga cgagaccggc
     1861 gaataccggt acagcatggg gtttcagtgt gagacagagg gcggctccgg aggcagcggc
     1921 ggttctggag gttccggtgg cggctccgga ggtagcggag ggtctcacac tctttacgcc
     1981 cctggaggat acgacattat gggatatttg attcagatta tgaaccgccc aaaccctcag
     2041 gtcgaactgg ggcctgtgga cacgtcatgt gccctgatcc tgtgcgatct gaagcaaaag
     2101 gacactccga ttgtctacgc ctcggaagcc ttcttgtata tgaccggata cagcaatgca
     2161 gaggtgctcg gtaggaactg cagattcctg cagtcccccg acgggatggt gaaaccaaag
     2221 tcgactcgca aatatgtgga ctcgaacacg atcaatacaa tgcggaaggc catcgaccgg
     2281 aacgccgagg tccaggtgga ggtggtcaac tttaagaaga acggccagcg gttcgtgaac
     2341 tttctcacca tgattccggt ccgggatgaa accggagagt acagatactc catgggattc
     2401 cagtgcgaaa ccgaagggtc cggaggtccc ggaggtgatg tggcagtgaa aatgttgaat
     2461 gtgacagcac ctacacctca gcagttacaa gccttcaaaa atgaagtagg agtactcagg
     2521 aaaacacgac atgtgaatat cctactcttc atgggctatt ccacaaagcc acaactggct
     2581 attgttaccc agtggtgtga gggctccagc ttgtatcacc atctccatat cattgagacc
     2641 aaatttgaga tgatcaaact tatagatatt gcacgacaga ctgcacaggg catggattac
     2701 ttacacgcca agtcaatcat ccacagagac ctcaagagta ataatatatt tcttcatgaa
     2761 gacctcacag taaaaatagg tgattttggt ctagctacag agaaatctcg atggagtggg
     2821 tcccatcagt ttgaacagtt gtctggatcc attttgtgga tggcaccaga agtcatcaga
     2881 atgcaagata aaaatccata cagctttcag tcagatgtat atgcatttgg aattgttctg
     2941 tatgaattga tgactggaca gttaccttat tcaaacatca acaacaggga ccagataatt
     3001 tttatggtgg gacgaggata cctgtctcca gatctcagta aggtacggag taactgtcca
     3061 aaagccatga agagattaat ggcagagtgc ctcaaaaaga aaagagatga gagaccactc
     3121 tttccccaaa ttctcgcctc tattgagctg ctggcccgct cattgccaaa aattcaccgc
     3181 agtgcatcag aaccctcctt gaatcgggct ggtttccaaa cagaggattt tagtctatat
     3241 gcttgtgctt ctccaaaaac acccatccag gcagggggat atggtgcgtt tcctgtccac
     3301 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg
     3361 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3421 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     3481 aagctgaccc tgaagttgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3541 gtgaccaccc tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     3601 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3661 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3721 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3781 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     3841 atcaaggcaa acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     3901 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3961 ctgagctacc agtccaaact gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4021 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caag
//
