LOCUS       sequence-133686-         831 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 831)
  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 831)
  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..831
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     85..104
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        184..259
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    307..424
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        493..704
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      724..733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             733..756
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             763..783
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
       61 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
      121 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggaagcat cggctgcgcc
      181 tgggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      241 agtggcaccg agtcggtgct tttttgaatt ctagatcttg agacaaatgg cagtattcat
      301 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
      361 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
      421 ttttcgggtt tattacaggg acagcagaga tccactttgg cgccggctcg agtggctccg
      481 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
      541 tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      601 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      661 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtgtcg tgacgcggga
      721 tccgccacca tggattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
      781 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct g
//
