LOCUS       sequence-27540-e         889 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 889)
  AUTHORS   Evers TH, van Dongen EM, Faesen AC, Meijer EW, Merkx M
  TITLE     Quantitative understanding of the energy transfer between 
            fluorescent proteins connected via flexible peptide linkers.
  JOURNAL   Biochemistry. 2006 Nov 7. 45(44):13183-92.
  PUBMED    17073440
REFERENCE   2  (bases 1 to 889)
  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"; journalName: 
            "Biochemistry. 2006 Nov 7. 45(44):13183-92."
COMMENT     Sequence Label: pET28CLY3
FEATURES             Location/Qualifiers
     source          1..889
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             3..25
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             44..61
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             71..88
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(92..94,95..97,98..807)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMXSCT"
     primer_bind     complement(137..158)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(398..417)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     745..766
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 attttgttta actttaagaa ggagatatac tatgggcagc agccatcatc atcatcatca
       61 cagcagcggc ctggtgccgc gcggcagcca tatggtgagc aagggcgagg agctgttcac
      121 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      181 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      241 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca
      301 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      361 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      421 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      481 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa
      541 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
      601 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      661 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
      721 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      781 cactctcggc atgncgagct gtacaagtcc ggaattcgtg gtggatccgg tggatctggt
      841 ggatccggtg gtagtggtgg atccggaggt agcaccatgg tgagcaagg
//
