LOCUS       sequence-127647-        1049 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR gRNA expression vector with an improved scaffold and puro/mAG
            markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1049)
  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 1049)
  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)-PGKpuro2AmAG-W
FEATURES             Location/Qualifiers
     source          1..1049
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             74..670
                     /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(74..93)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     570..590
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             689..742
                     /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 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc atccatctag atctcgagca
       61 gctgaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
      121 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
      181 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
      241 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
      301 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
      361 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
      421 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
      481 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
      541 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
      601 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
      661 gcccggtgcc ggcggcgggt ccggaggaga gggcagagga agtctcctaa catgcggtga
      721 cgtggaggag aatcctggcc cagaattcat ggtgagcgtg atcaagcccg agatgaagat
      781 caagctgtgc atgaggggca ccgtgaacgg ccacaacttc gtgatcgagg gcgagggcaa
      841 gggcaacccc tacgagggca cccagatcct ggacctgaac gtgaccgagg gcgcccccct
      901 gcccttcgcc tacgacatcc tgaccaccgt gttccagtac ggcaacaggg ccttcaccaa
      961 gtaccccgcc gacatccagg actacttcaa gcagaccttc cccgagggct accactggga
     1021 gaggagcatg acctacgagg accagggca
//
