LOCUS       sequence-146638-         901 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  ""All-in-one"" CRISPR/Cas9 (wt) plasmid for cloning of custom gRNA 
            with blue/white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 901)
  AUTHORS   Findlay SD, Vincent KM, Berman JR, Postovit LM
  TITLE     A Digital PCR-Based Method for Efficient and Highly Specific 
            Screening of Genome Edited Cells.
  JOURNAL   PLoS One. 2016 Apr 18;11(4):e0153901. doi: 
            10.1371/journal.pone.0153901. eCollection 2016.
  PUBMED    27089539
REFERENCE   2  (bases 1 to 901)
  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: "PLoS One.";
            date: "2016-04-18"; volume: "11(4):e0153901. doi"; pages: " 
            10.1371/journal.pone.0153901. eCollection 2016"
COMMENT     Sequence Label: All_in_one_CRISPR/Cas9_LacZ
FEATURES             Location/Qualifiers
     source          1..901
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             287..307
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             308..334
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             308..334
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(351..371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        356..685
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      536..671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     598..617
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     regulatory      743..752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcgcgtgatc ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg
       61 cgacaagccc atccgcgagc aggccgagaa catcatccac ctgttcaccc tgaccaacct
      121 gggcgccccc gccgccttca agtacttcga caccaccatc gaccgcaagc gctacaccag
      181 caccaaggag gtgctggacg ccaccctgat ccaccagagc atcaccggtc tgtacgagac
      241 ccgcatcgac ctgagccagc tgggcggcga cggcggctcc ggacctccaa agaaaaagag
      301 aaaagtatac ccctacgacg tgcccgacta cgcctaataa ctcgagctgt ggaatgtgtg
      361 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      421 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      481 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
      541 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
      601 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
      661 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
      721 tgatcggcgc gccataccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
      781 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
      841 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
      901 g
//
