LOCUS       sequence-109593-         848 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 848)
  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 848)
  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..848
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      28..37
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(56..88)
                     /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(92..109)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(119..136)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             155..177
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    192..216
                     /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(216..235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(217..235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          296..499
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     complement(319..339)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc ccgcgaccca
       61 tttgctgtcc accagtcatg ctagccatat ggctgccgcg cggcaccagg ccgctgctgt
      121 gatgatgatg atgatggctg ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt
      181 atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc
      241 gcgggatcga gatctgggcc cgcggtaccg tcgactgcag aattcctcac gacacctgaa
      301 atggaagaaa aaaactttga accactgtct gaggcttgag aatgaaccaa gatccaaact
      361 caaaaagggc aaattccaag gagaattaca tcaagtgcca agctggccta acttcagtct
      421 ccacccactc agtgtgggga aactccatcg cataaaaccc ctccccccaa cctaaagacg
      481 acgtactcca aaagctcgag aactaatcga ggtgcctgga cggcgcccgg tactccgtgg
      541 agtcacatga agcgacggct gaggacggaa aggccctttt cctttgtgtg ggtgactcac
      601 ccgcccgctc tcccgagcgc cgcgtcctcc attttgagct ccctgcagca gggccgggaa
      661 gcggccatct ttccgctcac gcaactggtg ccgaccgggc cagccttgcc gcccagggcg
      721 gggcgataca cggcggcgcg aggccaggca ccagagcagg ccggccagct tgagactacc
      781 cccgtccgat tctcggtggc cgcgctcgca ggccccgcct cgccgaacat gtgcgctggg
      841 acgcacgg
//
