LOCUS       sequence-133769-         914 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 914)
  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 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/nature14363"; journalName: "Nature"; date: "2015-04-16- 
            16"; volume: "520"; issue: "7547"; pages: "363-7"
COMMENT     Sequence Label: sgGLDC_2
FEATURES             Location/Qualifiers
     source          1..914
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     99..118
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        198..273
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    321..438
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        507..718
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      738..747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             747..770
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             777..797
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
       61 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
      121 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgatt
      181 tggggtagac atcgcccgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      241 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg aattctagat cttgagacaa
      301 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
      361 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
      421 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg
      481 ctcgagtggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
      541 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
      601 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
      661 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
      721 gtcgtgacgc gggatccgcc accatggatt acaaagacga tgacgataag atggccccaa
      781 agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag tacagcatcg
      841 gcctggncat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
      901 ccagcaagaa ttca
//
