LOCUS       sequence-288259-         825 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for sACE2-sfGFP high affinity variant 
            4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 825)
  AUTHORS   Chan KK, Dorosky D, Sharma P, Abbasi SA, Dye JM, Kranz DM, Herbert 
            AS, Procko E
  TITLE     Engineering human ACE2 to optimize binding to the spike protein of 
            SARS coronavirus 2.
  JOURNAL   Science. 2020 Sep 4;369(6508):1261-1265. doi: 
            10.1126/science.abc0870. Epub 2020 Aug 4.
  PUBMED    32753553
REFERENCE   2  (bases 1 to 825)
  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: 
            "10.1126/science.abc0870"; journalName: "Science"; date: "2020-09-4-
            4"; volume: "369"; issue: "6508"; pages: "1261-1265"
COMMENT     Sequence Label: pcDNA3-sACE2v4-sfGFP
FEATURES             Location/Qualifiers
     source          1..825
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(12..725)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             complement(15..725)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(55..75)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     670..698
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gactcgagtt atcatttgta gagctcatcc atgccatgtg taatcccagc agcagttaca
       61 aactcaagaa ggaccatgtg gtcacgcttt tcgttgggat ctttcgaaag gacagattgt
      121 gtcgacaggt aatggttgtc tggtaaaagg acagggccat cgccaattgg agtattttgt
      181 tgataatggt ctgctagttg aacggaacca tcttcaacgt tgtggcgaat tttgaagtta
      241 gctttgattc cattcttttg tttgtctgcc gtgatgtata cattgtgtga gttaaagttg
      301 tactccagtt tgtgtccgag aatgtttcca tcttctttaa aatcaatacc ttttaactcg
      361 atacgattaa caagggtatc accttcaaac ttgacttcag cacgcgtctt gtaggtcccg
      421 tcatctttga aagatatagt gcgttcctgt acataacctt cgggcatggc actcttgaaa
      481 aagtcatgcc gtttcatgtg atccggataa cgggaaaagc attgaacacc ataggtcaga
      541 gtagtgacaa gtgttggcca tggaacaggt agttttccag tagtgcaaat aaatttaagg
      601 gtgagttttc cgtttgtagc atcaccttca ccctctccac ggacagaaaa tttgtgccca
      661 ttaacatcac catctaattc aacaagaatt gggacaactc cagtgaaaag ttcttctcct
      721 ttggaaccac cactaccgga cccaccggat ccgtctgcat atggactcca gtcggtactc
      781 catcccacaa aagaattctt gttctggtct ttcagccagg taaat
//
