LOCUS       sequence-410240-        1551 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses cFOS-FRB-VP16 and EGFP component in rat hippocampal neuron
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1551)
  AUTHORS   Cui M, Lee S, Ban SH, Ryu JR, Shen M, Yang SH, Kim JY, Choi SK, Han 
            J, Kim Y, Han K, Lee D, Sun W, Kwon HB, Lee D
  TITLE     A single-component, light-assisted uncaging switch for 
            endoproteolytic release.
  JOURNAL   Nat Chem Biol. 2024 Mar;20(3):353-364. doi: 
            10.1038/s41589-023-01480-6. Epub 2023 Nov 16.
  PUBMED    37973890
REFERENCE   2  (bases 1 to 1551)
  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"; doi: 
            "10.1038/s41589-023-01480-6"; journalName: "Nat Chem Biol"; date: 
            "2024-03"; volume: "20"; issue: "3"; pages: "353-364"
COMMENT     Sequence Label: pAAV-cFOS-FRB-VP16-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..1551
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     13..32
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             91..369
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             526..759
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             769..825
                     /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"
     CDS             join(835..837,838..1551)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(835..837,838..1548)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(877..898)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1138..1157)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1485..1506
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcagccca
       61 agcggatcgg ccgccaccat ggcttctcga atcctctggc atgagatgtg gcatgaaggc
      121 ctggaagagg catctcgttt gtactttggg gaaaggaacg tgaaaggcat gtttgaggtg
      181 ctggagccct tgcatgctat gatggaacgg ggcccccaga ctctgaagga aacatccttt
      241 aatcaggcct atggtcgaga tttaatggag gcccaagagt ggtgcaggaa gtacatgaaa
      301 tcagggaatg tcaaggacct cctccaagcc tgggacctct attatcatgt gttccgacga
      361 atctcaaaga ctagtgcgta cagccgcgcg cgtacgaaaa acaattacgg gtctaccatc
      421 gagggcctgc tcgatctccc ggacgacgac gcccccgaag aggcggggct ggcggctccg
      481 cgcctgtcct ttctccccgc gggacacacg cgcagactat cgacggcccc cccgaccgat
      541 gtcagcctgg gggacgagct ccacttagac ggcgaggacg tggcgatggc gcatgccgac
      601 gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg tccgggattt
      661 accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga gtttgagcag
      721 atgtttaccg atgcccttgg aattgacgag tacggtgggg gaagcggagc tactaacttc
      781 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctgaatt cggggtgagc
      841 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      901 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      961 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1021 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1081 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1141 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1201 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1261 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1321 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1381 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1441 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1501 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg a
//
