LOCUS       sequence-121154-        1110 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses wild type PA-DAD in mammalian cells, an optogenetic tool 
            to activate endogenous diaphanous related formins in live cells 
            using light..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1110)
  AUTHORS   Rao MV, Chu PH, Hahn KM, Zaidel-Bar R
  TITLE     An optogenetic tool for the activation of endogenous 
            diaphanous-related formins induces thickening of stress fibers 
            without an increase in contractility.
  JOURNAL   Cytoskeleton (Hoboken). 2013 Jul;70(7):394-407. doi: 
            10.1002/cm.21115. Epub 2013 May 24.
  PUBMED    23677607
REFERENCE   2  (bases 1 to 1110)
  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"; doi: "10.1002/cm.21115"; 
            journalName: "Cytoskeleton (Hoboken)"; date: "2013-07"; volume: 
            "70"; issue: "7"; pages: "394-407"
COMMENT     Sequence Label: PA-DAD-wt
FEATURES             Location/Qualifiers
     source          1..1110
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    46..70
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        86..195
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             216..233
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(240..242,243..953)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(282..303)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(543..562)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     890..911
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggcaaagaat tggatcggac cgaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc ccggagttaa tccgggacct ttaattcaac ccaacacaat atattatagt
      121 taaataagaa ttattatcaa atcatttgta tattaattaa aatactatac tgtaaattac
      181 attttattta caatcaaagg agatatacca tggcacacca tcaccaccat cacggttctg
      241 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      301 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      361 agctgaccct gaagttcatc agcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      421 tgaccaccct cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      481 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      541 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      601 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      661 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
      721 tcaaggcaaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
      781 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
      841 tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
      901 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggttctg
      961 gaagtggatc cttggctact acacttgaac gtattgagaa gaactttgtc attactgacc
     1021 caagattgcc agataatccc attatattcg cgtccgatag tttcttgcag ttgacagaat
     1081 atagccgtga agaaattttg ggaagaaact
//
