LOCUS       Exported                1112 bp ds-DNA     linear   SYN 12-JUN-2021
DEFINITION  Expresses dark mutant PA-DAD in mammalian cells, an optogenetic tool
            to activate endogenous diaphanous related formins in live cells 
            using light with LOV2 in the closed conformation.
ACCESSION   .
VERSION     .
KEYWORDS    PA-DAD-dark
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1112)
  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 1112)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1112
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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        79..188
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             209..226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             233..946
                     /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(275..296)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(536..555)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     883..904
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aattggatcg gaccgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tccccggagt taatccggga cctttaattc aacccaacac aatatattat agttaaataa
      121 gaattattat caaatcattt gtatattaat taaaatacta tactgtaaat tacattttat
      181 ttacaatcaa aggagatata ccatggcaca ccatcaccac catcacggtt ctgtgagcaa
      241 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      301 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      361 cctgaagttc atcagcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      421 cctcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
      481 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      541 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      601 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      661 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
      721 aaacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
      781 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
      841 ccagtccaaa ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
      901 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtt ctggaagtgg
      961 atccttggct actacacttg aacgtattga gaagaacttt gtcattactg acccaagatt
     1021 gccagataat cccattatat tcgcgtccga tagtttcttg cagttgacag aatatagccg
     1081 tgaagaaatt ttgggaagaa acatgaggtt tc
//
