LOCUS       sequence-93072-e         882 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral Vector for Sp sgRNA delivery without 
            SpCas9, GFP, EFS Promoter driven.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 882)
  AUTHORS   Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey 
            ME, Thielke A, Aster JC, Regev A, Ebert BL
  TITLE     Generation of mouse models of myeloid malignancy with combinatorial 
            genetic lesions using CRISPR-Cas9 genome editing.
  JOURNAL   Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951.
  PUBMED    24952903
REFERENCE   2  (bases 1 to 882)
  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"; journalName: "Nat 
            Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951."
COMMENT     Sequence Label: pLKO5.sgRNA.EFS.GFP
FEATURES             Location/Qualifiers
     source          1..882
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(43..756,757..759,760..762))
                     /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(46..756,757..759,760..762))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(88..109)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     437..456
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     696..717
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      759..768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 acaaattttg taatccagag gttgattgtc gacttaacgc gtttacttgt acagctcgtc
       61 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
      121 ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag
      181 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
      241 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
      301 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
      361 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
      421 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
      481 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
      541 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
      601 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
      661 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
      721 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcgg atcccgcgtc
      781 acgacacctg tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg
      841 cacttatata cggttctccc ccaccctcgg gaaaaaggcg ga
//
