LOCUS       sequence-110721-         577 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 577)
  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 577)
  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..577
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    81..105
                     /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)."
     RBS             120..142
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             161..178
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             188..205
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             209..241
                     /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"
     regulatory      260..269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(311..332)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ggtgtcgtga ggaattctgc agtcgacggt accgcgggcc cagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
      121 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
      181 cagcggcctg gtgccgcgcg gcagccatat ggctagcatg actggtggac agcaaatggg
      241 tcgcggggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      301 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      361 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      421 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctagggcg tgcagtgctt
      481 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      541 ctacgtccag gagcgcacca tcttcttcaa ggacgac
//
