LOCUS       sequence-114692-        1004 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid for tet-inducible expression of dCas9-EGFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1004)
  AUTHORS   Ochiai H, Sugawara T, Yamamoto T
  TITLE     Simultaneous live imaging of the transcription and nuclear position 
            of specific genes.
  JOURNAL   Nucleic Acids Res. 2015 Jun 19. pii: gkv624.
  PUBMED    26092696
REFERENCE   2  (bases 1 to 1004)
  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: "Nucleic 
            Acids Res. 2015 Jun 19. pii: gkv624."
COMMENT     Sequence Label: pBSK-Delta-B-TRE-dCas9-EGFP
FEATURES             Location/Qualifiers
     source          1..1004
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(33..746,747..749,750..752))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(36..746,747..749,750..752))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(78..99)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     427..446
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     686..707
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(840..860)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggaattctta gctagcccat cagcattcta gatcacttgt acagctcgtc catgccgaga
       61 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
      121 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
      181 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
      241 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
      301 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
      361 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
      421 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
      481 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
      541 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
      601 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
      661 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
      721 ccggtgaaca gctcctcgcc cttgctcacc atggatccac ctccgcctga ccctccgcca
      781 ccgcttccgc cgcctccgag tgcggccagc gcggggtccg gcctcgagag gctggcctcc
      841 acctttctct tcttcttggg gctgtcgcct cccagctgag acaggtcgat ccgtgtctcg
      901 tacaggccgg tgatgctctg gtggatcagg gtggcgtcca gcacctcttt ggtgctggtg
      961 tacctcttcc ggtcgatggt ggtgtcaaag tacttgaagg cggc
//
