LOCUS       sequence-108077-         893 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized S. pyogenes Cas9 protein and EGFP 
            from EFS promoter. 3rd generation lentiviral backbone. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 893)
  AUTHORS   Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song 
            J, Pan JQ, Weissleder R, Lee H, Zhang F, Sharp PA
  TITLE     Genome-wide CRISPR screen in a mouse model of tumor growth and 
            metastasis.
  JOURNAL   Cell. 2015 Mar 12;160(6):1246-60. doi: 10.1016/j.cell.2015.02.038. 
            Epub 2015 Mar 5.
  PUBMED    25748654
REFERENCE   2  (bases 1 to 893)
  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.cell.2015.02.038"; journalName: "Cell"; date: 
            "2015-03-12- 12"; volume: "160"; issue: "6"; pages: "1246-60"
COMMENT     Sequence Label: lentiCas9-EGFP
FEATURES             Location/Qualifiers
     source          1..893
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(88..801,802..804))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(133..154)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     482..501
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     741..762
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(805..861)
                     /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             complement(871..885)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
       61 attaccgata agcttgatat cgaattctta cttgtacagc tcgtccatgc cgagagtgat
      121 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
      181 gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
      241 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
      301 gcggatcttg aagttcgcct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
      361 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
      421 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
      481 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
      541 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
      601 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
      661 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccagacac
      721 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
      781 gaacagctcc tcgcccttgc tcaccggtcc aggattctct tcgacatctc cggcttgttt
      841 cagcagagag aagtttgttg cgccggatcc cttatcgtca tcgtctttgt aat
//
