LOCUS       sequence-163213-         973 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  stable knockout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 973)
  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 973)
  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-sgSHMT1-G3
FEATURES             Location/Qualifiers
     source          1..973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     125..144
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        224..299
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    347..464
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        533..744
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      764..773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             773..796
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             803..823
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
       61 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      121 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      181 cttgtggaaa ggacgaaaca ccggaacggg gcgtatctca tgggttttag agctagaaat
      241 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      301 tttttgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      361 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      421 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      481 acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca gtgggcagag
      541 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      601 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      661 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      721 aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca tggattacaa
      781 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      841 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      901 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      961 ccggcacagc atc
//
