LOCUS       sequence-110712-         444 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of the Celeste (cyan) spectral variant of eGFP in 
            bacteria and in mammalian cells. Could be 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 444)
  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 444)
  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(W66)
FEATURES             Location/Qualifiers
     source          1..444
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     67..86
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        67..85
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    86..110
                     /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             125..147
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             166..183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             193..210
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             214..246
                     /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      265..274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(316..337)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 tttcaggtgt cgtgaggaat tctgcagtcg acggtaccgc gggcccagat ctcgatcccg
       61 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
      121 taattttgtt taactttaag aaggagatat accatgggca gcagccatca tcatcatcat
      181 cacagcagcg gcctggtgcc gcgcggcagc catatggcta gcatgactgg tggacagcaa
      241 atgggtcgcg gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      301 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      361 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      421 accggcaagc tgcccgtgcc ctgg
//
