LOCUS       sequence-135855-         887 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral dual CRISPR gRNA expression vector (human 7SK and human 
            U6 promoters).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 887)
  AUTHORS   Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, 
            Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi 
            S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli 
            P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly 
            BJ, Iorio F, Pina C, Vassiliou GS, Yusa K
  TITLE     A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and 
            Therapeutic Targets in Acute Myeloid Leukemia.
  JOURNAL   Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 
            10.1016/j.celrep.2016.09.079.
  PUBMED    27760321
REFERENCE   2  (bases 1 to 887)
  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.celrep.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-18- 18"; volume: "17"; issue: "4"; pages: "1193-1205"
COMMENT     Sequence Label: pKLV2.2-h7SKgRNA5(SapI)-hU6gRNA5(BbsI)-PGKpuro...
FEATURES             Location/Qualifiers
     source          1..887
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             71..667
                     /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(71..90)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     567..587
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             686..739
                     /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"
ORIGIN
        1 gttctcctct tcctcatctc cgggcctttc gacctgcatc catctagatc tcgagcagct
       61 gaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
      121 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
      181 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
      241 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
      301 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
      361 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
      421 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
      481 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
      541 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
      601 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
      661 cggtgccggc ggcgggtccg gaggagaggg cagaggaagt ctcctaacat gcggtgacgt
      721 ggaggagaat cctggcccaa tgagcgagct gattaaggag aacatgcaca tgaagctgta
      781 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc
      841 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggcc
//
