LOCUS       sequence-135181-        1097 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized S. pyogenes Cas9 protein and Venus 
            reporter from EFS promoter. Lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1097)
  AUTHORS   Canver MC, Smith EC, Sher F, Pinello L, Sanjana NE, Shalem O, Chen 
            DD, Schupp PG, Vinjamur DS, Garcia SP, Luc S, Kurita R, Nakamura Y, 
            Fujiwara Y, Maeda T, Yuan GC, Zhang F, Orkin SH, Bauer DE
  TITLE     BCL11A enhancer dissection by Cas9-mediated in situ saturating 
            mutagenesis.
  JOURNAL   Nature. 2015 Sep 16. doi: 10.1038/nature15521.
  PUBMED    26375006
REFERENCE   2  (bases 1 to 1097)
  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: "Nature. 
            2015 Sep 16. doi: 10.1038/nature15521."
COMMENT     Sequence Label: lentiCas9-Venus
FEATURES             Location/Qualifiers
     source          1..1097
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(90..803,804..806,807..809))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(135..156)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     484..503
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     743..764
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(810..866)
                     /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(876..899)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(900..947)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 taaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
       61 tgattatcga taagcttgat atcgaattct tacttgtaca gctcgtccat gccgagagtg
      121 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
      181 ttgctcaggg cggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
      241 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
      301 tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
      361 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
      421 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
      481 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
      541 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
      601 tgcaggccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
      661 gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
      721 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
      781 gtgaacagct cctcgccctt gctcaccatc ggtccaggat tctcttcgac atctccggct
      841 tgtttcagca gagagaagtt tgttgcgccg gatcccttat cgtcatcgtc tttgtaatct
      901 ttcttcttct tagcctgtcc agccttcttt gtggcggcag gtcgcttgtc gcctcccagc
      961 tgagacaggt cgatccgtgt ctcgtacagg ccggtgatgc tctggtggat cagggtggcg
     1021 tccagcacct ctttggtgct ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg
     1081 aaggcggcag gggctcc
//
