LOCUS       sequence-182227-         936 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  TEV::eGFP::ER::Ollas::linker::3XFlag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 936)
  AUTHORS   Paix A, Folkmann A, Seydoux G
  TITLE     Precision genome editing using CRISPR-Cas9 and linear repair 
            templates in C. elegans.
  JOURNAL   Methods. 2017 May 15;121-122:86-93. doi: 
            10.1016/j.ymeth.2017.03.023. Epub 2017 Apr 7.
  PUBMED    28392263
REFERENCE   2  (bases 1 to 936)
  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: "Methods."; 
            date: "2017-05-15"; volume: "121-122:86-93. doi"; pages: " 
            10.1016/j.ymeth.2017.03.023. Epub 2017 Apr 7"
COMMENT     Sequence Label: pAP684-3
FEATURES             Location/Qualifiers
     source          1..936
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             16..36
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(37..39,40..750)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(67..95)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     690..710
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             763..804
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             856..921
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cggtacccgg ggatcgagaa cctctacttc caaggagtga gtaaaggaga agaacttttc
       61 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
      121 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
      181 actactggaa aactacctgt tccatggcca acacttgtca ctactctgac ctacggcgtg
      241 cagtgcttca gccggtaccc cgaccacatg aagcagcatg actttttcaa gagtgccatg
      301 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
      361 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
      421 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
      481 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
      541 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
      601 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
      661 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
      721 attacactcg gcatggacga gctgtacaag tacgtagccg gctccggatt cgccaacgag
      781 ctcggaccac gtctcatggg aaagtgcccg ggggatcggt ggagctccac cggtggcggc
      841 cgctctagaa ctagtgacta caaagaccat gacggtgatt ataaagatca tgatatcgat
      901 tacaaggatg acgatgacaa ggatcctcta gagtcg
//
