LOCUS       sequence-160831-        1239 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  stable knockout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1239)
  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 1239)
  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-sgAAVS1
FEATURES             Location/Qualifiers
     source          1..1239
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     146..165
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        244..319
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    367..484
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        553..764
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      784..793
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             793..816
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             823..843
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cttcatctgc tatacgatac aaggctgtta gagagtaatt agaattaatt tgactgtaaa
       61 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      121 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      181 cgatttcttg gctttatata tcttgtggaa aggacgaaac accggggcca ctagggacag
      241 gatgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      301 agtggcaccg agtcggtgct tttttgaatt ctagatcttg agacaaatgg cagtattcat
      361 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
      421 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
      481 ttttcgggtt tattacaggg acagcagaga tccactttgg cgccggctcg agtggctccg
      541 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
      601 tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      661 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      721 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtgtcg tgacgcggga
      781 tccgccacca tggattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
      841 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
      901 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
      961 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     1021 tcgacagcgg cgaaacagcg agccacccgc tgagagaccg cagaagaaga tacacagacg
     1081 agacgatctg ctatctgcaa gagatctcag cacggatgca gttgacgaag ctctccanga
     1141 ctgagagtct ctgtggagag atagagcnac gaaccaactc gnaacttctg nagtgctacn
     1201 gagatcacat taactgagac tgtggagcca cgccaagnc
//
