LOCUS       sequence-223221-        1797 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 sgRNA2 
            against the EGFP CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1797)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 1797)
  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-guide2
FEATURES             Location/Qualifiers
     source          1..1797
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     28..47
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        127..202
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        264..475
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      488..497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             524..709
                     /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 ttgnagtttt aaattatgtt ttaaatggac tatcatatgc ttaccgtaac ttgaaagtat
       61 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggagc tggacggcga
      121 cgtaaagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      181 aaaagtggca ccgagtcggt gcttttttga attcgctagc taggtcttga aaggagtggg
      241 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
      301 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
      361 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
      421 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg
      481 gtgatccgcc accatggatg ctaagtcact aactgcctgg tcccggacac tggtgacctt
      541 caaggatgta tttgtggact tcaccaggga ggagtggaag ctgctggaca ctgctcagca
      601 gatcgtgtac agaaatgtga tgctggagaa ctataagaac ctggtttcct tgggttatca
      661 gcttactaag ccagatgtga tcctccggtt ggagaaggga gaagagcccg gcggttccgg
      721 cggagggtcg atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
      781 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
      841 caccgaccgg cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggaga
      901 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
      961 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     1021 cttccacaga ctggaagagt ccttcctggg ngaaaaagga taagaagcac gaacgggcac
     1081 cccatcttcg gcaacatcgt ggaacaaggt ggcctaccac gagaagtacc ccaccatcta
     1141 ccacctgaag aaaaaaactg ggtggaaagc caccgaacag ggcccaaact gcgggctgaa
     1201 tctatttggg ccctgggccc caatgaaaca agtttccggg ggccactttc ccggaatcaa
     1261 ggggcgaacc tggacccccg gaaaaaaggc ggacgtggta aaaaactggt ttattccccc
     1321 tgtgttggaa aaacatcaaa anccancggn gtcgaaggaa aaacccccct caagncccnc
     1381 ggggggtggg gaaccaccag ggccacctct tgtttcgcgc atttagngaa naaaacaaaa
     1441 aagnctgtgg aaaaatttgt atccccncat ctcctccgnc ganaaaaaaa aaataagggc
     1501 tgtgngtcgc acccncatag ngttcccgng caaaggcggg gggcggcgac acacccnatt
     1561 ataagannaa atttccttac cctgggggga agggagtggc cattcggcct gttntaaaga
     1621 agagacacct cccaacacag aacaacccag ngtacccaag ctggcgnggc tccagagtnt
     1681 tcgggaaaca caaaaccccc ttatgtttgt gtgcccgccg caacaaacct agacccccga
     1741 ctcctggnga tggaaacctg gnnaaagaaa gggaaaaccg naccccggcc ccnnttt
//
