LOCUS       sequence-92777-e         914 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  An empty gRNA expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 914)
  AUTHORS   Koike-Yusa H, Li Y, Tan EP, Velasco-Herrera Mdel C, Yusa K
  TITLE     Genome-wide recessive genetic screening in mammalian cells with a 
            lentiviral CRISPR-guide RNA library.
  JOURNAL   Nat Biotechnol. 2014 Mar;32(3):267-73. doi: 10.1038/nbt.2800. Epub 
            2013 Dec 23.
  PUBMED    24535568
REFERENCE   2  (bases 1 to 914)
  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: "10.1038/nbt.2800"; 
            journalName: "Nat Biotechnol"; date: "2014-03"; volume: "32"; issue:
            "3"; pages: "267-73"
COMMENT     Sequence Label: pKLV-U6gRNA(BbsI)-PGKpuro2ABFP
FEATURES             Location/Qualifiers
     source          1..914
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             65..661
                     /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(65..84)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     561..581
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             680..733
                     /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 ctcttcctca tctccgggcc tttcgacctg catccatcta gatctcgagc agctgaagct
       61 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
      121 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
      181 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
      241 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
      301 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
      361 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
      421 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
      481 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
      541 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
      601 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
      661 cggcggcggg tccggaggag agggcagagg aagtctccta acatgcggtg acgtggagga
      721 gaatcctggc ccaatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
      781 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
      841 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
      901 catcctggct acta
//
