LOCUS       sequence-223215-        1423 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Catalytically inactive Cas9 from S. pyogenes with P2A-EGFP 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 1423)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 1423)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..1423
                     /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             429..482
                     /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"
     CDS             join(483..485,486..998)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIE"
     primer_bind     complement(525..546)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(786..805)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 nnnnnntnnn gnntntnnag 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 gggatcccga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
      481 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      541 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      601 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      661 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      721 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      781 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      841 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      901 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
      961 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1021 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1081 acctgagcac ccagtccgcc ctgaacaaag accccaacga gaagcgcgat cacatggtcc
     1141 tggctggaat tcgtgaccgc cgcccgggat nnctcncggc atggacaanc tgtacaagga
     1201 attctaancg nnttaannga caatcaaccn tnggnattnc aaaattgtgg aagaaatgan
     1261 gggggattct taacannntn gcccntttta cccnagtggn aaacccngct tnanagccct
     1321 tgtttaagcc nnntggttcc cccaaggggt tnttttttct ccccntgaaa aaacccggnt
     1381 ncggncctct ttnnnnnaan nntgggcccc ttncntgnaa agg
//
