LOCUS       sequence-124759-         947 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 activity reporter with BFP and GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 947)
  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 947)
  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(gGFP)-PGKBFP2AGFP-W
FEATURES             Location/Qualifiers
     source          1..947
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(77..790,791..793,794..796))
                     /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(80..790,791..793,794..796))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(122..143)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     471..490
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     730..751
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(803..856)
                     /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 gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca
       61 gaggttgagc ggccgcttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
      121 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
      181 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
      241 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
      301 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
      361 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
      421 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
      481 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
      541 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
      601 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
      661 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
      721 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
      781 cgcccttgct caccatgaat tcaggtccgg ggttctcttc cacgtcgccg caggtgagca
      841 gggagcccct tccttccccg ccgctgcccc ctccattaag cttgtgcccc agtttgctag
      901 ggaggtcgca gtatctggcc actgccacct cgtgctgctc gacgtag
//
