LOCUS       sequence-322256-        1621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression and localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1621)
  AUTHORS   Miserey-Lenkei S, Trajkovic K, D'Ambrosio JM, Patel AJ, Copic A, 
            Mathur P, Schauer K, Goud B, Albanese V, Gautier R, Subra M, Kovacs 
            D, Barelli H, Antonny B
  TITLE     A comprehensive library of fluorescent constructs of SARS-CoV-2 
            proteins and their initial characterisation in different cell types.
  JOURNAL   Biol Cell. 2021 Jul;113(7):311-328. doi: 10.1111/boc.202000158. Epub
            2021 May 10.
  PUBMED    33666950
REFERENCE   2  (bases 1 to 1621)
  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: 
            "10.1111/boc.202000158"; journalName: "Biol Cell"; date: "2021-07"; 
            volume: "113"; issue: "7"; pages: "311-328"
COMMENT     Sequence Label: ORF3a-EGFP
FEATURES             Location/Qualifiers
     source          1..1621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      5..14
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    836..888
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      896..905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(902..904,905..907,908..1621)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(902..904,905..907,908..1618)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(947..968)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1208..1227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1555..1576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctagccacc atggaccttt ttatgagaat ctttaccatc gggactgtca cactgaaaca
       61 gggagaaatt aaggacgcga ccccgagtga ctttgtgagg gccactgcta caattcccat
      121 ccaagcctct cttccattcg gctggctcat agtgggtgtg gcactgctcg cggtattcca
      181 gtcagcgtct aagatcataa cgctgaagaa gagatggcag ttggctctca gcaaaggtgt
      241 tcatttcgtc tgtaacctgc ttcttctttt cgtcactgtc tattctcact tgctcttggt
      301 tgcagccggg ctggaagccc cgtttctgta tctgtacgcg ctcgtttact tcctccagag
      361 cataaatttc gtacgaatta ttatgagact ctggttgtgc tggaagtgtc ggagcaagaa
      421 tcctctcttg tatgacgcca attacttcct ttgttggcac actaattgtt atgattactg
      481 cataccttac aatagcgtta cctcatccat agtaatcacc tccggtgacg gaacaacgtc
      541 accaatctcc gagcatgatt accagattgg aggctatacg gaaaaatggg agagtggtgt
      601 caaggactgt gtggtgctgc atagctattt cacaagcgat tattatcaac tgtacagtac
      661 acagttgtcc acggataccg gggtcgagca tgtcacattt tttatttata acaagatcgt
      721 tgatgagcca gaggagcatg tgcagatcca cacgatagat ggaagctccg gtgtagtgaa
      781 tccagtcatg gaaccgatct atgacgagcc caccacgacc acatctgtgc ccctgctcga
      841 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac
      901 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1081 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1381 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1501 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1621 a
//
