LOCUS       sequence-223211-        1298 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Catalytically inactive Cas9 from S. pyogenes with P2A-BlastR under 
            the EF1a core promoter, and cloning backbone for sgRNA. Contains 
            BsmBI sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1298)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 1298)
  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
FEATURES             Location/Qualifiers
     source          1..1298
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             294..320
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             300..326
                     /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="YDVPDYASL"
     CDS             339..359
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             366..386
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             432..488
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(968..988)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 aanntnngcg agttctccag agagtgatcc tggccgacgc taatctggac aaggtgctga
       61 gcgcctacaa caagcacaga gacaagccta tcagagagca ggccgagaat atcatccacc
      121 tgtttaccct gaccagtctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
      181 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
      241 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac gcctatccct
      301 atgacgtgcc cgattatgcc agcctgggca gcggctcccc caagaaaaaa cgcaaggtgg
      361 aagatcctaa gaaaaagcgg aaagtggacg gcattggtag tgggagcaac ggcagcagcg
      421 gggatccggc cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc gaagagaatc
      481 ctggaccgat ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
      541 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
      601 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
      661 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
      721 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
      781 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
      841 ttgggattcg tgaattgctg ccctctggtt angttgtggg aagggctaan aaattcgata
      901 tcaagcttat cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
      961 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcagg
     1021 ctattgcttc ccgaagggct ttcattttcc ccccctggaa aaaacccggg tggcggctcc
     1081 tttnntannn nnnntnggcc gcttttcaag caacaaggcc ccggntnggc cacttggttt
     1141 gttgaaccaa ccccccccgg gtgtggggga tggccccccc cggggagccc ctttcnngga
     1201 gatttntttt tccccccccc attgnngggc ggagacaatt nccccccccc tttccccctc
     1261 ttagagaaag ggcgcggttg tgggcttaga aatacaac
//
