LOCUS       sequence-163490-        1039 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  stable knockout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1039)
  AUTHORS   Pacold ME, Brimacombe KR, Chan SH, Rohde JM, Lewis CA, Swier LJ, 
            Possemato R, Chen WW, Sullivan LB, Fiske BP, Cho S, Freinkman E, 
            Birsoy K, Abu-Remaileh M, Shaul YD, Liu CM, Zhou M, Koh MJ, Chung H,
            Davidson SM, Luengo A, Wang AQ, Xu X, Yasgar A, Liu L, Rai G, 
            Westover KD, Vander Heiden MG, Shen M, Gray NS, Boxer MB, Sabatini 
            DM
  TITLE     A PHGDH inhibitor reveals coordination of serine synthesis and 
            one-carbon unit fate.
  JOURNAL   Nat Chem Biol. 2016 Jun;12(6):452-8. doi: 10.1038/nchembio.2070. 
            Epub 2016 Apr 25.
  PUBMED    27110680
REFERENCE   2  (bases 1 to 1039)
  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/nchembio.2070"; journalName: "Nat Chem Biol"; date: 
            "2016-06"; volume: "12"; issue: "6"; pages: "452-8"
COMMENT     Sequence Label: pLentiCRISPRv1-sgPHGDH-G5
FEATURES             Location/Qualifiers
     source          1..1039
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     131..150
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        230..305
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    353..470
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        539..750
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      770..779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             779..802
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             809..829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
       61 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      121 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      181 atatatcttg tggaaaggac gaaacaccgg cttctgccag accaatccag ttttagagct
      241 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      301 ggtgcttttt tgaattctag atcttgagac aaatggcagt attcatccac aattttaaaa
      361 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
      421 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
      481 acagggacag cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg
      541 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
      601 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
      661 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
      721 tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga
      781 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
      841 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
      901 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
      961 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     1021 aacagccgag gccacccgg
//
