LOCUS       sequence-127883-        1044 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses TEV::eGFP::myc::3Xflag in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1044)
  AUTHORS   Paix A, Folkmann A, Rasoloson D, Seydoux G
  TITLE     High Efficiency, Homology-Directed Genome Editing in Caenorhabditis 
            elegans Using CRISPR-Cas9 Ribonucleoprotein Complexes.
  JOURNAL   Genetics. 2015 Sep;201(1):47-54. doi: 10.1534/genetics.115.179382. 
            Epub 2015 Jul 17.
  PUBMED    26187122
REFERENCE   2  (bases 1 to 1044)
  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"; doi: 
            "10.1534/genetics.115.179382"; journalName: "Genetics"; date: 
            "2015-09"; volume: "201"; issue: "1"; pages: "47-54"
COMMENT     Sequence Label: AP682-1
FEATURES             Location/Qualifiers
     source          1..1044
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             18..38
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(39..41,42..752)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(69..97)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     692..712
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             765..794
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             846..911
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(959..975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(959..975)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(972..994)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    983..999
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1007..1013,1014..1031,1032..1037))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ctcggtaccc ggggatcgag aacctctact tccaaggagt gagtaaagga gaagaacttt
       61 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
      121 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
      181 gcactactgg aaaactacct gttccatggc caacacttgt cactactctg acctacggcg
      241 tgcagtgctt cagccggtac cccgaccaca tgaagcagca tgactttttc aagagtgcca
      301 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
      361 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
      421 ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac
      481 acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac ttcaaaatta
      541 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
      601 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
      661 cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg
      721 ggattacact cggcatggac gagctgtaca agtacgtagc cggcgaacaa aaactgatat
      781 cagaagagga tctgtgcccg ggggatcggt ggagctccac cggtggcggc cgctctagaa
      841 ctagtgacta caaagaccat gacggtgatt ataaagatca tgatatcgat tacaaggatg
      901 acgatgacaa ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt
      961 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1021 gaagcataaa gtgtaaagcc tggg
//
