LOCUS       sequence-223223-        1656 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPRi negative control. Catalytically inactive Cas9 from S. 
            pyogenes with 2A-BlastR under the EF1a core promoter, and sgRNA3 
            against the EGFP CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1656)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 1656)
  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"
COMMENT     Sequence Label: Lenti-(BB)-EF1a-KRAB-dCas9-P2A-BlastR EGFP-guide3
FEATURES             Location/Qualifiers
     source          1..1656
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     30..49
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        129..204
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        266..477
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      490..499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             526..711
                     /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="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
ORIGIN
        1 tnnttgnagt tttaattatg ttttaaatgg actatcatat gcttaccgta acttgaaagt
       61 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgga tgccgttctt
      121 ctgcttgtgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      181 gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta gctaggtctt gaaaggagtg
      241 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
      301 ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg
      361 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
      421 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggac
      481 cggtgatccg ccaccatgga tgctaagtca ctaactgcct ggtcccggac actggtgacc
      541 ttcaaggatg tatttgtgga cttcaccagg gaggagtgga agctgctgga cactgctcag
      601 cagatcgtgt acagaaatgt gatgctggag aactataaga acctggtttc cttgggttat
      661 cagcttacta agccagatgt gatcctccgg ttggagaagg gagaagagcc cggcggttcc
      721 ggcggagggt cgatggacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg
      781 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
      841 aacaccgacc ggcacagcat caagaagaac ctgatcggcg ccctgctgtt cgacagcgga
      901 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
      961 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     1021 ttcttccaaa gactggaaga gtccttcctg gtggaaaaag gataaaaagc acaaccggca
     1081 ccccatcttc ggcaacatcc tggaacaagg tggcctaccc ccaaaaagta cccccacctt
     1141 ctaccacctg gaaaagaaaa ctgggtggaa cagcaccgaa caaggcccaa cctgggggct
     1201 gaatctaatt gggccctggg cccaaatgaa acaagtttcc gggggccctt tctcgaatta
     1261 aaggggcaac cttgaacccc ggaaaaaacc ggcaacgtgg gaaaaaactt gttatccacc
     1321 gcttggtggc aaacncttca accaaccgtt gttcgaagan gaaaaccctc ttacannccc
     1381 gcgagggggg tggggaaccg cacaggccct cctccgggtc ncgccaacan atnnganaaa
     1441 gaaccaaacg ctggtggaaa aatcaattag cccccccctt ctctcgcgnc gaagaaaaat
     1501 anngattngc gttttgtggc ganncacctg tanatgtccc cgggnantcg ggggcgtggg
     1561 acacccccac cttntttnga aaaggccttn tccctgccgc ctcgggaaag aagtntantc
     1621 atctctntng ctggaagaag aaaccagccn atttta
//
