LOCUS       Exported                 863 bp ds-DNA     linear   SYN 23-MAY-2021
DEFINITION  Photoactivatable transcription system. Lentiviral expression of 
            ASCL1 sgRNA1. Also contains a CMV-puro-t2A-mCherry expression 
            cassette..
ACCESSION   .
VERSION     .
KEYWORDS    sgRNA1_ASCL1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 863)
  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 863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 23, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..863
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      113..122
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(129..182)
                     /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(192..788)
                     /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(272..292)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     769..788
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(822..860)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
       61 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatggtg
      121 gcgaccggtg ggccaggatt ctcctcgacg tcaccgcatg ttagcagact tcctctgccc
      181 tctcccgatc cggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
      241 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
      301 gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
      361 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
      421 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
      481 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
      541 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
      601 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
      661 atgtggcggt ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
      721 agggtgcgta cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
      781 tcggtcatgg tgggctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc
      841 tcccaccgta cacgcctacc gcc
//
