LOCUS       sequence-125680-        1000 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR gRNA expression vector with an improved l scaffold and 
            puro/BFP markers without WPRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1000)
  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 1000)
  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-U6gRNA5(BbsI)-PGKpuro2ABFP
FEATURES             Location/Qualifiers
     source          1..1000
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             73..669
                     /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(73..92)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     569..589
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             688..741
                     /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 ctgttctcct cttcctcatc tccgggcctt tcgacctgca tccatctaga tctcgagcag
       61 ctgaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
      121 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
      181 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
      241 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
      301 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
      361 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
      421 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
      481 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
      541 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
      601 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
      661 cccggtgccg gcggcgggtc cggaggagag ggcagaggaa gtctcctaac atgcggtgac
      721 gtggaggaga atcctggccc aatgagcgag ctgattaagg agaacatgca catgaagctg
      781 tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag
      841 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
      901 gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc
      961 cagggcatcc ccgacttctt caagcagtcc ttccctgagg
//
