LOCUS       sequence-158737-         900 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  3rd generation lentiviral vector expressing GFP alongside Cas9 and 
            an sgRNA cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 900)
  AUTHORS   Walter DM, Venancio OS, Buza EL, Tobias JW, Deshpande C, Gudiel AA, 
            Kim-Kiselak C, Cicchini M, Yates TJ, Feldser DM
  TITLE     Systematic in vivo inactivation of chromatin regulating enzymes 
            identifies Setd2 as a potent tumor suppressor in lung 
            adenocarcinoma.
  JOURNAL   Cancer Res. 2017 Feb 15. pii: canres.2159.2016. doi: 
            10.1158/0008-5472.CAN-16-2159.
  PUBMED    28202515
REFERENCE   2  (bases 1 to 900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer Res.
            2017 Feb 15. pii: canres.2159.2016. doi: 
            10.1158/0008-5472.CAN-16-2159."
COMMENT     Sequence Label: LentiCRISPRv2GFP
FEATURES             Location/Qualifiers
     source          1..900
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             43..90
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             91..114
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             124..180
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(181..183,184..186,187..897)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(226..247)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(487..506)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     834..855
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaagcgacc tgccgccaca
       61 aagaaggctg gacaggctaa gaagaagaaa gattacaaag acgatgacga taagggatcc
      121 ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
      181 atggtgtcta agggcgaaga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      241 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      301 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      361 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      421 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      481 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      541 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      601 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      661 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      721 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      781 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      841 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
//
