LOCUS       sequence-109519-         868 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Photoactivatable transcription system. Lentiviral expression of 
            NANOG sgRNA2. Also contains a CMV-puro-t2A-mCherry expression 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 868)
  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 868)
  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: sgRNA2_NANOG
FEATURES             Location/Qualifiers
     source          1..868
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      103..112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(119..172)
                     /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(182..778)
                     /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(262..282)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     759..778
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(812..863)
                     /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(841..861)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga
       61 actccttgat gatggccatg ttatcctcct cgcccttgct caccatggtg gcgaccggtg
      121 ggccaggatt ctcctcgacg tcaccgcatg ttagcagact tcctctgccc tctcccgatc
      181 cggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
      241 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
      301 ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc
      361 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
      421 ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac
      481 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
      541 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
      601 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
      661 ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
      721 cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg
      781 tgggctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta
      841 cacgcctacc gcccatttgc gtcaatgg
//
