LOCUS       sequence-125666-         948 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 activity reporter (control) with BFP and GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 948)
  AUTHORS   Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, 
            Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi 
            S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli 
            P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly 
            BJ, Iorio F, Pina C, Vassiliou GS, Yusa K
  TITLE     A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and 
            Therapeutic Targets in Acute Myeloid Leukemia.
  JOURNAL   Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 
            10.1016/j.celrep.2016.09.079.
  PUBMED    27760321
REFERENCE   2  (bases 1 to 948)
  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: 
            "10.1016/j.celrep.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-18- 18"; volume: "17"; issue: "4"; pages: "1193-1205"
COMMENT     Sequence Label: pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W
FEATURES             Location/Qualifiers
     source          1..948
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(65..778,779..781,782..784))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(68..778,779..781,782..784))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(110..131)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     459..478
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     718..739
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(791..844)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
ORIGIN
        1 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgagcgg
       61 ccgcttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
      121 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
      181 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
      241 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
      301 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
      361 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
      421 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
      481 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
      541 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
      601 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
      661 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
      721 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
      781 ccatgaattc aggtccgggg ttctcttcca cgtcgccgca ggtgagcagg gagccccttc
      841 cttccccgcc gctgccccct ccattaagct tgtgccccag tttgctaggg aggtcgcagt
      901 atctggccac tgccacctcg tgctgctcga cgtaggtctc gttgttgg
//
