LOCUS       sequence-347364-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing K417N-L452R-T478K 
            mutations with single chain Fc, HRV3C protease cleavage site, and 
            AVI tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  AUTHORS   Teng IT, Nazzari AF, Choe M, Liu T, Oliveira de Souza M, Petrova Y, 
            Tsybovsky Y, Wang S, Zhang B, Artamonov M, Madan B, Huang A, Lopez 
            Acevedo SN, Pan X, Ruckwardt TJ, DeKosky BJ, Mascola JR, Misasi J, 
            Sullivan NJ, Zhou T, Kwong PD
  TITLE     Molecular probes of spike ectodomain and its subdomains for 
            SARS-CoV-2 variants, Alpha through Omicron.
  JOURNAL   PLoS One. 2022 May 24;17(5):e0268767. doi: 
            10.1371/journal.pone.0268767. eCollection 2022.
  PUBMED    35609088
REFERENCE   2  (bases 1 to 6558)
  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: "PLoS One.";
            date: "2022-05-24"; volume: "17(5):e0268767. doi"; pages: " 
            10.1371/journal.pone.0268767. eCollection 2022"
COMMENT     Sequence Label: pVRC8400-SARS-CoV-2-RBD-K417N-L452R-T478K-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(570..1385)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(570..1385)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1293..1312
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1377..1396
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1663..1681
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1719..1741)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1841..1860
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2011..2390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2391..2594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2545..2565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2591..2615
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4524..4547
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5169..5213
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5236..5460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5827..5844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5998..6558,1..28))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6078..6097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
       61 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      121 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      181 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      241 tctatttcgt tcatccatag ttgcctgact cggggggggg gggcgctgag gtctgcctcg
      301 tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt
      361 gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt
      421 ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc
      481 agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc
      541 cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
      601 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
      661 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
      721 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
      781 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
      841 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
      901 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
      961 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1021 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1081 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1141 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1201 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1261 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1321 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg
     1381 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1441 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttccccc
     1501 cccccccatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1561 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1621 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     1681 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1741 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1801 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1861 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     1921 atcagattgg ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata
     1981 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
     2041 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2101 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2161 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2221 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2281 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2341 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2401 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2461 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2521 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2581 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     2641 ttgacctcca tagaagacac cgggaccgat ccagcctcca tcggctcgca tctctccttc
     2701 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     2761 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     2821 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     2881 gctttgcctg accctgcttg ctcaactcta gttaacggtg gagggcagtg tagtctgagc
     2941 agtactcgtt gctgccgcgc gcgccaccag acataatagc tgacagacta acagactgtt
     3001 cctttccatg ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag atatcgcggc
     3061 cgctctagac caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg
     3121 gtgtacattc cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc
     3181 ctaaggacac cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt
     3241 cccacgagga ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg
     3301 ccaagaccaa gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga
     3361 cagtgctgca ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg
     3421 ccctgcctgc cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc
     3481 aggtgtatac cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact
     3541 gcctggtgaa gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc
     3601 cagagaacaa ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt
     3661 attccaagct gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg
     3721 tgatgcacga ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca
     3781 agggaggagg aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca
     3841 gcgcccctga actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata
     3901 ccctgatgat ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg
     3961 accccgaagt caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca
     4021 agcctagaga agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc
     4081 atcaggattg gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg
     4141 cccctatcga gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata
     4201 cactgccacc cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca
     4261 agggcttcta ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca
     4321 attataaaac taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc
     4381 tgacagtgga taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg
     4441 aggccctgca taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca
     4501 gcggaggcgg aggcagcgga ggcctggagg tgctgtttca gggccctcca aacattacca
     4561 acctctgccc cttcggcgag gtgtttaacg ccacaagatt tgcctccgtt tacgcctgga
     4621 atagaaagag aatcagcaat tgtgtggccg actactccgt gctgtataac agcgcctctt
     4681 tcagcacctt caagtgctac ggcgtttccc caacaaagct gaatgacctg tgcttcacca
     4741 acgtgtacgc cgactccttc gtaattagag gcgatgaggt gcggcagatc gcaccaggcc
     4801 agaccggtaa catcgctgac tacaactata agctgcctga tgattttaca ggctgcgtga
     4861 tcgcctggaa ctctaacaac ctggatagca aggtgggcgg caactacaac taccggtacc
     4921 ggctgtttcg caagtctaac ctgaaacctt tcgagagaga catctccaca gagatctacc
     4981 aggccggttc taagccttgt aacggggtgg aaggcttcaa ctgttacttc cctctgcaaa
     5041 gctacggctt ccagcctacc aatggagtcg gctaccagcc ataccgggtg gtcgtgctgt
     5101 ccttcgagtt actccacgcc cccgccaccg tctgcggtgg aggaggcctg gtgccacagc
     5161 agagcggagg cctgaacgac atcttcgagg cccagaagat cgagtggcac gagggctgat
     5221 gaggatccag atctgctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5281 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5341 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5401 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5461 gtacccaggt gctgaagaat tgacccggtt cctcctgggc cagaaagaag caggcacatc
     5521 cccttctctg tgacacaccc tgtccacgcc cctggttctt agttccagcc ccactcatag
     5581 gacactcata gctcaggagg gctccgcctt caatcccacc cgctaaagta cttggagcgg
     5641 tctctccctc cctcatcagc ccaccaaacc aaacctagcc tccaagagtg ggaagaaatt
     5701 aaagcaagat aggctattaa gtgcagaggg agagaaaatg cctccaacat gtgaggaagt
     5761 aatgagagaa atcatagaat tttaaggcca tgatttaagg ccatcatggc cttaatcttc
     5821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6541 ttttgtttgc aagcagca
//
