LOCUS       sequence-361466-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain from the BA.2 variant 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-BA.2-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(92..907)
                     /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(92..907)
                     /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     815..834
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     899..918
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1185..1203
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1241..1263)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1363..1382
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1533..1912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1913..2116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2067..2087
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2113..2137
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4046..4069
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4691..4735
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    4758..4982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5349..5366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5520..6108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5600..5619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct
       61 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
      121 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
      181 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
      241 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
      301 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
      361 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
      421 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
      481 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
      541 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
      601 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
      661 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
      721 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
      781 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
      841 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
      901 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
      961 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttncc
     1021 ccccccccca ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1141 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1201 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1261 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1321 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     1381 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     1441 gcatcagatt ggctattggc cattgcatac gttgtatcca tatcataata tgtacattta
     1501 tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata
     1561 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     1621 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1681 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1741 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1801 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1861 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1921 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1981 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2041 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2101 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     2161 ttttgacctc catagaagac accgggaccg atccagcctc catcggctcg catctctcct
     2221 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     2281 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     2341 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     2401 acgctttgcc tgaccctgct tgctcaactc tagttaacgg tggagggcag tgtagtctga
     2461 gcagtactcg ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg
     2521 ttcctttcca tgggtctttt ctgcagtcac cgtcgtcgac acgtgtgatc agatatcgcg
     2581 gccgctctag accaccatgg gatggtcatg tatcatcctt tttctagtag caactgcaac
     2641 cggtgtacat tccgcaccag agctgctggg aggacctagc gtgttcctgt ttccccctaa
     2701 gcctaaggac accctgatga tcagccggac cccagaggtg acatgcgtgg tggtggacgt
     2761 gtcccacgag gaccccgagg tgaagttcaa ctggtacgtg gatggcgtgg aggtgcacaa
     2821 tgccaagacc aagccaaggg aggagcagta caactctacc tatcgcgtgg tgagcgtgct
     2881 gacagtgctg caccaggact ggctgaacgg caaggagtac aagtgtaagg tgtctaataa
     2941 ggccctgcct gccccaatcg agaagaccat cagcaaggca aagggacagc caagggagcc
     3001 tcaggtgtat accctgccac ccagccgcga cgagctgaca aagaaccagg tgtccctgta
     3061 ctgcctggtg aagggcttct atccttccga tatcgccgtg gagtgggagt ctaatggcca
     3121 gccagagaac aattacaaga ccacacctcc agtgctggac tccgatggct ctttctttct
     3181 gtattccaag ctgaccgtgg ataagagccg gtggcagcag ggcaacgtgt ttagctgttc
     3241 cgtgatgcac gaggccctgc acaatcacta cacacagaag tctctgagcc tgtcccccgg
     3301 caagggagga ggaggcagcg gcggaggagg ctccggaggc ggcggctctg gcggcggcgg
     3361 cagcgcccct gaactgctgg gaggaccaag cgtgttcctg tttccaccta agcccaaaga
     3421 taccctgatg atctccagga cccctgaagt cacctgcgtg gtcgtggacg tgtctcatga
     3481 ggaccccgaa gtcaagttca actggtacgt ggacggagtc gaagtccata acgccaagac
     3541 caagcctaga gaagagcagt acaactccac ctatagagtg gtgtctgtgc tgacagtgct
     3601 gcatcaggat tggctgaatg gcaaggaata taagtgtaag gtgagcaata aggccctgcc
     3661 cgcccctatc gagaagacca tctctaaagc caaaggacag ccaagggagc cacaggtgta
     3721 tacactgcca cccagccggg acgagctgac caagaaccag gtgagcctga catgtctggt
     3781 caagggcttc taccccagtg acattgctgt ggagtgggag tccaatggcc agcctgagaa
     3841 caattataaa actaccccac cagtgctgga cagcgatggc tccttctttc tgaccagcaa
     3901 gctgacagtg gataagtccc gctggcagca gggaaacgtg ttttcttgta gcgtgatgca
     3961 tgaggccctg cataaccact acacccagaa gtccctgtct ctgagcccag gcaagggagg
     4021 cagcggaggc ggaggcagcg gaggcctgga ggtgctgttt cagggccctc caaacattac
     4081 caacctctgc cccttcgatg aggtgtttaa cgccacaaga tttgcctccg tttacgcctg
     4141 gaatagaaag agaatcagca attgtgtggc cgactactcc gtgctgtata actttgcccc
     4201 gttcttcgcc ttcaagtgct acggcgtttc cccaacaaag ctgaatgacc tgtgcttcac
     4261 caacgtgtac gccgactcct tcgtaattag aggcaatgag gtgagccaga tcgcaccagg
     4321 ccagaccggt aacatcgctg actacaacta taagctgcct gatgatttta caggctgcgt
     4381 gatcgcctgg aactctaaca aactggatag caaggtgggc ggcaactaca actacctgta
     4441 ccggctgttt cgcaagtcta acctgaaacc tttcgagaga gacatctcca cagagatcta
     4501 ccaggccggt aataaacctt gtaacggggt ggcaggcttc aactgttact tccctctgcg
     4561 tagctacggc ttccgtccta cctatggagt cggccaccag ccataccggg tggtcgtgct
     4621 gtccttcgag ttactccacg cccccgccac cgtctgcggt ggaggaggcc tggtgccaca
     4681 gcagagcgga ggcctgaacg acatcttcga ggcccagaag atcgagtggc acgagggctg
     4741 atgaggatcc agatctgctg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4801 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4861 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4921 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4981 gggtacccag gtgctgaaga attgacccgg ttcctcctgg gccagaaaga agcaggcaca
     5041 tccccttctc tgtgacacac cctgtccacg cccctggttc ttagttccag ccccactcat
     5101 aggacactca tagctcagga gggctccgcc ttcaatccca cccgctaaag tacttggagc
     5161 ggtctctccc tccctcatca gcccaccaaa ccaaacctag cctccaagag tgggaagaaa
     5221 ttaaagcaag ataggctatt aagtgcagag ggagagaaaa tgcctccaac atgtgaggaa
     5281 gtaatgagag aaatcataga attttaaggc catgatttaa ggccatcatg gccttaatct
     5341 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5401 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5461 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5521 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5581 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5641 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5701 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5761 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5821 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5881 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5941 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6001 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6061 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6121 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6181 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6241 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6301 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact cgggggnggg
     6361 gggcgctgag gtctgcctcg tgaagaaggt gttgctgact cataccaggc ctgaatcgcc
     6421 ccatcatcca gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac
     6481 cagttggtga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc
     6541 gtgatctgat ccttcaac
//
