LOCUS       sequence-109588-         859 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Photoactivatable transcription system. Lentiviral expression of 
            sgRNA3 to target GAL4UAS-luciferase. Also contains a 
            CMV-puro-t2A-mCherry expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 859)
  AUTHORS   Nihongaki Y, Yamamoto S, Kawano F, Suzuki H, Sato M
  TITLE     CRISPR-Cas9-based Photoactivatable Transcription System.
  JOURNAL   Chem Biol. 2015 Feb 19;22(2):169-74. doi: 
            10.1016/j.chembiol.2014.12.011. Epub 2015 Jan 22.
  PUBMED    25619936
REFERENCE   2  (bases 1 to 859)
  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: 
            "10.1016/j.chembiol.2014.12.011"; journalName: "Chem Biol"; date: 
            "2015-02-19- 19"; volume: "22"; issue: "2"; pages: "169-74"
COMMENT     Sequence Label: sgRNA3_GAL4UAS-Luciferase reporter
FEATURES             Location/Qualifiers
     source          1..859
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      98..107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(114..167)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(177..773)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(257..277)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     754..773
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(807..858)
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(836..856)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc
       61 ttgatgatgg ccatgttatc ctcctcgccc ttgctcacca tggtggcgac cggtgggcca
      121 ggattctcct cgacgtcacc gcatgttagc agacttcctc tgccctctcc cgatccggca
      181 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
      241 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag
      301 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
      361 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
      421 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
      481 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
      541 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
      601 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccggg
      661 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
      721 cgggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catggtgggc
      781 tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc
      841 ctaccgccca tttgcgtca
//
