LOCUS       sequence-135783-         803 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR-Cas9 induced gene disruption.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 803)
  AUTHORS   Kim D, Fiske BP, Birsoy K, Freinkman E, Kami K, Possemato RL, 
            Chudnovsky Y, Pacold ME, Chen WW, Cantor JR, Shelton LM, Gui DY, 
            Kwon M, Ramkissoon SH, Ligon KL, Kang SW, Snuderl M, Vander Heiden 
            MG, Sabatini DM
  TITLE     SHMT2 drives glioma cell survival in ischaemia but imposes a 
            dependence on glycine clearance.
  JOURNAL   Nature. 2015 Apr 16;520(7547):363-7. doi: 10.1038/nature14363. Epub 
            2015 Apr 8.
  PUBMED    25855294
REFERENCE   2  (bases 1 to 803)
  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/nature14363"; journalName: "Nature"; date: "2015-04-16- 
            16"; volume: "520"; issue: "7547"; pages: "363-7"
COMMENT     Sequence Label: sgGCAT_2
FEATURES             Location/Qualifiers
     source          1..803
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     124..143
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        223..298
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    346..463
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        532..743
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      763..772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             772..795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
       61 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
      121 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
      181 ttgtggaaag gacgaaacac cggaagcatc ggctgcgcct gggttttaga gctagaaata
      241 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      301 ttttgaattc tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
      361 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
      421 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
      481 cagcagagat ccactttggc gccggctcga gtggctccgg tgcccgtcag tgggcagagc
      541 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
      601 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
      661 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
      721 acgggtttgc cgccagaaca caggtgtcgt gacgcgggat ccgccaccat ggattacaaa
      781 gacgatgacg ataagatggc ccc
//
