LOCUS       sequence-82638-e         897 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  serves as PCR-template to generate HR donors for C-terminal GFP-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 897)
  AUTHORS   Bottcher R, Hollmann M, Merk K, Nitschko V, Obermaier C, 
            Philippou-Massier J, Wieland I, Gaul U, Forstemann K
  TITLE     Efficient chromosomal gene modification with CRISPR/cas9 and 
            PCR-based homologous recombination donors in cultured Drosophila 
            cells.
  JOURNAL   Nucleic Acids Res. 2014 Apr 19.
  PUBMED    24748663
REFERENCE   2  (bases 1 to 897)
  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. 2014 Apr 19."
COMMENT     Sequence Label: pMH3
FEATURES             Location/Qualifiers
     source          1..897
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(7..9,10..12,13..726)
                     /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             join(7..9,10..12,13..723)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(52..73)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(313..332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     660..681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(733..766)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ctcgagatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
       61 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      121 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      181 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      241 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      301 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      361 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      421 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
      481 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
      541 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      601 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
      661 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      721 aagtaaacta gtgaagttcc tatactttct agagaatagg aacttcaaat aaacatatgc
      781 tgttggaata tactattcaa cctacaaaaa taacgttaaa caacactact ttatatttga
      841 tatgaatggc cacacctttt atgccataaa acatattgta agagaatacc actcttt
//
