LOCUS       sequence-106129-         947 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lenti vector encoding dCAS9-VP64 with 2A Blast 
            resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 947)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
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"; journalName: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: lenti dCAS-VP64_Blast
FEATURES             Location/Qualifiers
     source          1..947
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             105..125
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             150..299
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             321..374
                     /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"
     CDS             375..773
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(851..871)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac agcgctggag
       61 gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag aaaaagagga
      121 aggtggcggc cgctggatcc ggacgggctg acgcattgga cgattttgat ctggatatgc
      181 tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg
      241 actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga
      301 ttaactgtac aggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
      361 agaatcctgg cccaatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
      421 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
      481 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
      541 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
      601 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
      661 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
      721 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagaattcg
      781 atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
      841 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
      901 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaa
//
