LOCUS       sequence-66585-e         909 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 909)
  AUTHORS   Orlowska KP, Klosowska K, Szczesny RJ, Cysewski D, Krawczyk PS, 
            Dziembowski A
  TITLE     A new strategy for gene targeting and functional proteomics using 
            the DT40 cell line.
  JOURNAL   Nucleic Acids Res. 2013 Sep;41(17):e167. doi: 10.1093/nar/gkt650. 
            Epub 2013 Jul 27.
  PUBMED    23892402
REFERENCE   2  (bases 1 to 909)
  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."; date: "2013-09"; volume: "41(17):e167. doi"; pages: " 
            10.1093/nar/gkt650. Epub 2013 Jul 27"
COMMENT     Sequence Label: pQuantEGFP-hygro-kana
FEATURES             Location/Qualifiers
     source          1..909
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(89..122)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             168..188
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(195..197,198..200,201..908)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(195..197,198..200,201..908)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(240..261)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(501..520)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     848..869
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctaacta attgagatgc
       61 atgctttgca tacttctgcc tgctggaaat aacttcgtat agcatacatt atacgaacgg
      121 tactcgaggg gaagcttgcc gccgatatca ccagcctgta caagggcgag aatttgtatt
      181 ttcagggtga gctcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      241 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      301 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      361 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      421 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      481 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      541 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      601 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      661 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
      721 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
      781 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
      841 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
      901 agctgtaca
//
