LOCUS       sequence-241888-        1632 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes lentivirus expression of L. monocytogenes CRISPR Type I-B 
            Flag-Cas6-KRAB-2a-BlastR driven by hUbC promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1632)
  AUTHORS   Pickar-Oliver A, Black JB, Lewis MM, Mutchnick KJ, Klann TS, 
            Gilcrest KA, Sitton MJ, Nelson CE, Barrera A, Bartelt LC, Reddy TE, 
            Beisel CL, Barrangou R, Gersbach CA
  TITLE     Targeted transcriptional modulation with type I CRISPR-Cas systems 
            in human cells.
  JOURNAL   Nat Biotechnol. 2019 Sep 23. pii: 10.1038/s41587-019-0235-7. doi: 
            10.1038/s41587-019-0235-7.
  PUBMED    31548729
REFERENCE   2  (bases 1 to 1632)
  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. 2019 Sep 23. pii: 10.1038/s41587-019-0235-7. doi: 
            10.1038/s41587-019-0235-7."
COMMENT     Sequence Label: pLenti-LmoCas6-KRAB-2a-BlastR
FEATURES             Location/Qualifiers
     source          1..1632
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..27
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             34..54
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             787..834
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             868..1062
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             1129..1149
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1159..1212
                     /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"
ORIGIN
        1 atggactaca aggacgacga cgataagatg gcgccaaaga aaaagcgcaa ggtccgcgga
       61 atgcggctga agattaactg cgacttcgac agcaagatca tccccaagga cttccagtcc
      121 aaggtcgtgt ccctgttcaa agccggaatt atgaaaagct ccccggagag atacgagaac
      181 ctgtttggtg gaaacaagca caagcagtac actttctcgg tgtatctccc aaaacctcag
      241 aacaagggcg ccgaaattca gctgaacgag gcgaactgta tcatcaactt ctccaccggc
      301 gatgctgaaa ccggtattat cttctacaac gccctgatta gcctgagagg ctccaaggtc
      361 ctgtttgggg ccggaaatca catcaccgtg aagaacatcc agatcgtgcc tgagaagaag
      421 atcatcggga agcaaaccat tctgcggacc ctttcgccgg tggtgtcgag ggatcacaac
      481 cgggaaacct tcaagaattg gttctactca ttcgaggacg aagaattcga gcctactttg
      541 aagcggaaca tgctgccgta cctcatcgac gcctttggag aacaggctcg ctacgatctg
      601 gaaaagctga agatcacgcc catttccatg aagaaagtcg tggtgtactg ccatgagatc
      661 cacatcgaga gctccgtggg catcttcgag ctggaagccg aggcatacct ccaaaagtac
      721 ctcgtggaaa acggaatcgg cactatgact ggctcagggt tcggaatgat tgaacagttc
      781 cggggcaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatcc
      841 gatgctaagt cactgactgc ctggtcccgg acactggtga ccttcaagga tgtgtttgtg
      901 gacttcacca gggaggagtg gaagctgctg gacactgctc agcagatcct gtacagaaat
      961 gtgatgctgg agaactataa gaacctggtt tccttgggtt atcagcttac taagccagat
     1021 gtgatcctcc ggttggagaa gggagaagag ccctggctgg tggagagaga aattcaccaa
     1081 gagacccatc ctgattcaga gactgcattt gaaatcaaat catcagttcc gaaaaagaaa
     1141 cgcaaagttg gaagcggaga gggcagagga agtcttctca catgcggtga cgtggaggag
     1201 aatcctggac ctaagacctt caacatctct cagcaggatc tggagctggt ggaggtcgcc
     1261 actgagaaga tcaccatgct ctatgaggac aacaagcacc atgtcggggc ggccatcagg
     1321 accaagactg gggagatcat ctctgctgtc cacattgagg cctacattgg cagggtcact
     1381 gtctgtgctg aagccattgc cattgggtct gctgtgagca acgggcagaa ggactttgac
     1441 accattgtgg ctgtcaggca cccctactct gatgaggtgg acagatccat cagggtggtc
     1501 agcccctgtg gcatgtgcag agagctcatc tctgactatg ctcctgactg ctttgtgctc
     1561 attgagatga atggcaagct ggtcaaaacc accattgagg aactcatccc cctcaagtac
     1621 accaggaact aa
//
