LOCUS       sequence-182232-         954 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3XFlag::TEV::6XHis::mTurquoise::6XHis::TEV::3XFlag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 954)
  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 954)
  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: pAP639-2F
FEATURES             Location/Qualifiers
     source          1..954
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             16..81
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             82..102
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             103..120
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(121..123,124..834)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(424..443)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     771..792
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             835..852
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             853..873
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             874..939
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cggtacccgg ggatcgatta taaagatcat gatggggatt ataaagatca tgatatagat
       61 tataaagatg atgatgataa agaaaattta tattttcagg gtcatcacca tcaccatcac
      121 gtgagcaagg gcgaggagct gttcaccgga gtggtgccca tcctggtcga gctggacgga
      181 gacgtaaacg gccacaagtt cagcgtgtcc ggagagggcg agggcgatgc cacctacggc
      241 aagctgaccc tcaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      301 gtgaccaccc tgtcctgggg cgtgcaatgc ttcgcccgct accccgacca catgaagcag
      361 cacgacttct tcaagtccgc catgcccgaa ggatacgtcc aggagcgcac catcttcttc
      421 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggaga caccctggtg
      481 aaccgcatcg agctgaaggg aatcgacttc aaggaggacg gcaacatcct ggggcacaag
      541 ctggagtaca actacatcag cgacaacgtc tacatcaccg ccgacaagca gaagaacggc
      601 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
      661 cactaccagc agaacacccc catcggagac ggccccgtgc tgctccccga caaccactac
      721 ctgagcaccc agtccaagct gtccaaagac cccaacgaga agcgcgatca catggtcctg
      781 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcaccat
      841 caccatcacc atgagaacct ctacttccaa ggagactaca aagaccatga cggtgattat
      901 aaagatcatg atatcgatta caaggatgac gatgacaagg atcctctaga gtcg
//
