LOCUS       sequence-322260-        2317 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 2317)
  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 2317)
  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: Nsp14D-EGFP
FEATURES             Location/Qualifiers
     source          1..2317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1532..1584
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1592..1601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1598..1600,1601..1603,1604..2317)
                     /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(1598..1600,1601..1603,1604..2314)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1643..1664)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1904..1923)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2251..2272
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctagccacc atgcaagcac ctacccacct ttccgtagat acgaagttca aaacggaagg
       61 attgtgtgtg gatataccag ggataccaaa ggatatgacg taccgaaggc tgatttccat
      121 gatgggtttt aagatgaatt accaagttaa tggctacccc aacatgttca tcaccaggga
      181 ggaggcaatt agacacgtaa gagcctggat aggcttcgat gttgaaggtt gccatgcaac
      241 ccgggaagcc gtaggcacaa accttccgtt gcagcttggc ttttccacgg gcgtcaacct
      301 cgttgccgta ccgactggct atgttgacac gccgaacaac accgatttct ctcgcgtaag
      361 tgctaagcct cctccgggag atcaattcaa gcatcttata cctctcatgt acaagggact
      421 gccgtggaat gtagttcgca ttaagatcgt gcagatgctc tccgatactt tgaaaaatct
      481 gagcgacagg gtggtcttcg ttctgtgggc acatggcttc gaacttactt ctatgaaata
      541 ctttgtaaaa atcgggcccg agcggacatg ttgtttgtgc gaccgacgag ctacttgttt
      601 tagcacagca tctgacacct acgcatgctg gcaccacagt ataggcttcg attacgtcta
      661 caatcccttt atgatagacg tacaacaatg gggctttacg ggtaacttgc agagtaatca
      721 cgatctttac tgccaagttc atgggaacgc acacgtggcc tcctgcgatg cgataatgac
      781 gaggtgcttg gccgtgcacg agtgctttgt taagcgggtc gattggacta tagagtatcc
      841 cataatcggt gacgaactta aaattaatgc tgcatgcagg aaagtgcagc acatggtagt
      901 aaaagcagcg ctcctcgcgg ataagtttcc tgtgctccac gatatcggca accctaaggc
      961 cattaaatgc gtcccccaag ctgatgtaga gtggaaattc tatgacgctc agccatgtag
     1021 tgacaaagca tacaagattg aagaattgtt ttattcatat gcgacacact ccgataagtt
     1081 cactgacgga gtttgtctct tttggaactg taatgttgat aggtatccgg cgaatagcat
     1141 agtgtgtcga tttgacacgc gcgttctctc taatttgaac ctccctggct gcgacggggg
     1201 gtctttgtac gtaaacaagc atgcatttca taccccagct tttgacaaat ccgctttcgt
     1261 aaatctgaaa caacttccct tcttctatta ttcagatagc ccttgcgaat ctcacggtaa
     1321 gcaagtggtt agtgacattg actatgtacc acttaaaagt gcaacgtgca ttacacgatg
     1381 caatttgggg ggcgcggttt gccgacacca tgccaacgaa tacagattgt acctggatgc
     1441 atataacatg atgataagtg cagggtttag tctttgggtg tataagcagt tcgatacgta
     1501 taatctctgg aacacattta cccgattgca gctcgagctc aagcttcgaa ttctgcagtc
     1561 gacggtaccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg gcgaggagct
     1621 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1681 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1741 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1801 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1861 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1921 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1981 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2041 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2101 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2161 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     2221 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2281 cgggatcact ctcggcatgg acgagctgta caagtaa
//
