LOCUS       sequence-199455-         993 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mammalian eGFP with TEV::3xFlag::Ollas::Myc::V5::HAtag::other 
            linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 993)
  AUTHORS   Paix A, Folkmann A, Goldman DH, Kulaga H, Grzelak MJ, Rasoloson D, 
            Paidemarry S, Green R, Reed RR, Seydoux G
  TITLE     Precision genome editing using synthesis-dependent repair of 
            Cas9-induced DNA breaks.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114.
  PUBMED    29183983
REFERENCE   2  (bases 1 to 993)
  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: "Proc Natl 
            Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 
            10.1073/pnas.1711979114."
COMMENT     Sequence Label: pAP2042-1
FEATURES             Location/Qualifiers
     source          1..993
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..21
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             22..87
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             88..129
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             130..159
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             160..201
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             202..228
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(280..282,283..993)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(280..282,283..993)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(322..343)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(583..602)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     930..951
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gagaaccttt attttcaggg agactataag gaccatgacg gggattataa agatcacgac
       61 attgattaca aggacgacga cgataagtcc ggcttcgcta acgaattggg gccaaggctc
      121 atgggcaagg aacagaaatt gatctctgag gaggacctgg gtaaacctat cccaaatccc
      181 ctgttgggtt tggactcaac ttacccctac gacgttcctg actatgcttg ccctggtgac
      241 cgctggagtt caacaggagg cggcagatca agaacgagtg tgagcaaggg cgaggagctg
      301 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      361 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      421 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
      481 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      541 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      601 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      661 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      721 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
      781 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
      841 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
      901 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      961 gggatcactc tcggcatgga cgagctgtac aag
//
