LOCUS       sequence-110725-         765 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of a non-sense mutant version of eGFP (dark) in bacteria 
            and in mammalian cells. Could be used as a reporter for gene 
            targeting and recombineering in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 765)
  AUTHORS   Valledor M, Hu Q, Schiller P, Myers RS
  TITLE     Fluorescent protein engineering by in vivo site-directed 
            mutagenesis.
  JOURNAL   IUBMB Life. 2012 Aug;64(8):684-9. doi: 10.1002/iub.1041. Epub 2012 
            May 28.
  PUBMED    22639380
REFERENCE   2  (bases 1 to 765)
  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: "10.1002/iub.1041"; 
            journalName: "IUBMB Life"; date: "2012-08"; volume: "64"; issue: 
            "8"; pages: "684-9"
COMMENT     Sequence Label: pDual-eGFP(Stop66)
FEATURES             Location/Qualifiers
     source          1..765
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     222..243
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      285..294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(313..345)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(349..366)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(376..393)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             412..434
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    449..473
                     /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)."
     primer_bind     complement(473..492)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(474..492)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          553..756
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     complement(576..596)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
       61 ggggtagcgg ctgaagcact gcacgcccta ggtcagggtg gtcacgaggg tgggccaggg
      121 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
      181 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
      241 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
      301 tggatccccg cgacccattt gctgtccacc agtcatgcta gccatatggc tgccgcgcgg
      361 caccaggccg ctgctgtgat gatgatgatg atggctgctg cccatggtat atctccttct
      421 taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag
      481 tgagtcgtat taatttcgcg ggatcgagat ctgggcccgc ggtaccgtcg actgcagaat
      541 tcctcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat
      601 gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc
      661 tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc
      721 cccccaacct aaagacgacg tactccaaaa gctcgagaac taatc
//
