LOCUS       sequence-108225-         831 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of eGFP in bacteria and in mammalian cells. Used as a 
            reporter for quantifying gene targeting and recombineering in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 831)
  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 831)
  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
FEATURES             Location/Qualifiers
     source          1..831
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     92..111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        92..110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    111..135
                     /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             150..172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             191..208
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             218..235
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             239..271
                     /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      290..299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(296..298,299..301,302..814)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(341..362)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(602..621)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattctgc agtcgacggt
       61 accgcgggcc cagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      121 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
      181 gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat
      241 ggctagcatg actggtggac agcaaatggg tcgcggggat ccaccggtcg ccaccatggt
      301 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      361 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      421 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      481 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
      541 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      601 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      661 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
      721 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
      781 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc t
//
