LOCUS       sequence-142631-         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Can clone in a C2H2 zinc finger via BsmBI restriction sites and 
            monitor post-translational degradation using EGFP:mCherry ratio 
            (Lentiviral, PGK.BsmBICloneSite-EGFP.IRES.mCherry.cppt.EF1a.PuroR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 886)
  AUTHORS   Slabicki M, Kozicka Z, Petzold G, Li YD, Manojkumar M, Bunker RD, 
            Donovan KA, Sievers QL, Koeppel J, Suchyta D, Sperling AS, Fink EC, 
            Gasser JA, Wang LR, Corsello SM, Sellar RS, Jan M, Gillingham D, 
            Scholl C, Frohling S, Golub TR, Fischer ES, Thoma NH, Ebert BL
  TITLE     The CDK inhibitor CR8 acts as a molecular glue degrader that 
            depletes cyclin K.
  JOURNAL   Nature. 2020 Sep;585(7824):293-297. doi: 10.1038/s41586-020-2374-x. 
            Epub 2020 Jun 3.
  PUBMED    32494016
REFERENCE   2  (bases 1 to 886)
  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/s41586-020-2374-x"; journalName: "Nature"; date: "2020-09";
            volume: "585"; issue: "7824"; pages: "293-297"
COMMENT     Sequence Label: Cilantro 2
FEATURES             Location/Qualifiers
     source          1..886
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      61..70
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(115..117,118..831)
                     /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(115..117,118..828)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(157..178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(418..437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     765..786
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cctcgttgac cgaatcaccg acctctctcc ccagggatcg ataccgtcga ctccggaata
       61 gccaccatgg agacggacgt ctcaggcggc ggcggctccg gcggcggcgg ctccgtgagc
      121 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      181 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      241 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      301 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      361 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      421 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      481 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      541 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
      601 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
      661 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
      721 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
      781 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aatgcatgag
      841 taactgagga tccgcccctc tccctccccc ccccctaacg ttactg
//
