LOCUS Exported 2317 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION mammalian expression and localization. ACCESSION . VERSION . KEYWORDS Nsp14D-EGFP 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 characterization in different cell types JOURNAL bioRxiv 423586 REFERENCE 2 (bases 1 to 2317) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2317 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1532..1584 /label=MCS /note="multiple cloning site" regulatory 1592..1601 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1598..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 1598..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 //