LOCUS       sequence-347375-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing 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-L452R-T478K-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..64)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     164..183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        334..713
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        714..917
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     868..888
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     914..938
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2847..2870
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3492..3536
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3559..3783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4150..4167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4321..4909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4401..4420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5451..6266)
                     /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(5451..6266)
                     /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     6174..6193
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6258..6277
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     join(6544..6558,1..4)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
       61 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      121 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      181 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      241 cgcatcagat tggctattgg ccattgcata cgttgtatcc atatcataat atgtacattt
      301 atattggctc atgtccaaca ttaccgccat gttgacattg attattgact agttattaat
      361 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      421 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      481 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      541 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      601 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      661 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      721 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      781 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      841 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      901 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
      961 gttttgacct ccatagaaga caccgggacc gatccagcct ccatcggctc gcatctctcc
     1021 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc
     1081 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag
     1141 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc
     1201 cacgctttgc ctgaccctgc ttgctcaact ctagttaacg gtggagggca gtgtagtctg
     1261 agcagtactc gttgctgccg cgcgcgccac cagacataat agctgacaga ctaacagact
     1321 gttcctttcc atgggtcttt tctgcagtca ccgtcgtcga cacgtgtgat cagatatcgc
     1381 ggccgctcta gaccaccatg ggatggtcat gtatcatcct ttttctagta gcaactgcaa
     1441 ccggtgtaca ttccgcacca gagctgctgg gaggacctag cgtgttcctg tttcccccta
     1501 agcctaagga caccctgatg atcagccgga ccccagaggt gacatgcgtg gtggtggacg
     1561 tgtcccacga ggaccccgag gtgaagttca actggtacgt ggatggcgtg gaggtgcaca
     1621 atgccaagac caagccaagg gaggagcagt acaactctac ctatcgcgtg gtgagcgtgc
     1681 tgacagtgct gcaccaggac tggctgaacg gcaaggagta caagtgtaag gtgtctaata
     1741 aggccctgcc tgccccaatc gagaagacca tcagcaaggc aaagggacag ccaagggagc
     1801 ctcaggtgta taccctgcca cccagccgcg acgagctgac aaagaaccag gtgtccctgt
     1861 actgcctggt gaagggcttc tatccttccg atatcgccgt ggagtgggag tctaatggcc
     1921 agccagagaa caattacaag accacacctc cagtgctgga ctccgatggc tctttctttc
     1981 tgtattccaa gctgaccgtg gataagagcc ggtggcagca gggcaacgtg tttagctgtt
     2041 ccgtgatgca cgaggccctg cacaatcact acacacagaa gtctctgagc ctgtcccccg
     2101 gcaagggagg aggaggcagc ggcggaggag gctccggagg cggcggctct ggcggcggcg
     2161 gcagcgcccc tgaactgctg ggaggaccaa gcgtgttcct gtttccacct aagcccaaag
     2221 ataccctgat gatctccagg acccctgaag tcacctgcgt ggtcgtggac gtgtctcatg
     2281 aggaccccga agtcaagttc aactggtacg tggacggagt cgaagtccat aacgccaaga
     2341 ccaagcctag agaagagcag tacaactcca cctatagagt ggtgtctgtg ctgacagtgc
     2401 tgcatcagga ttggctgaat ggcaaggaat ataagtgtaa ggtgagcaat aaggccctgc
     2461 ccgcccctat cgagaagacc atctctaaag ccaaaggaca gccaagggag ccacaggtgt
     2521 atacactgcc acccagccgg gacgagctga ccaagaacca ggtgagcctg acatgtctgg
     2581 tcaagggctt ctaccccagt gacattgctg tggagtggga gtccaatggc cagcctgaga
     2641 acaattataa aactacccca ccagtgctgg acagcgatgg ctccttcttt ctgaccagca
     2701 agctgacagt ggataagtcc cgctggcagc agggaaacgt gttttcttgt agcgtgatgc
     2761 atgaggccct gcataaccac tacacccaga agtccctgtc tctgagccca ggcaagggag
     2821 gcagcggagg cggaggcagc ggaggcctgg aggtgctgtt tcagggccct ccaaacatta
     2881 ccaacctctg ccccttcggc gaggtgttta acgccacaag atttgcctcc gtttacgcct
     2941 ggaatagaaa gagaatcagc aattgtgtgg ccgactactc cgtgctgtat aacagcgcct
     3001 ctttcagcac cttcaagtgc tacggcgttt ccccaacaaa gctgaatgac ctgtgcttca
     3061 ccaacgtgta cgccgactcc ttcgtaatta gaggcgatga ggtgcggcag atcgcaccag
     3121 gccagaccgg taagatcgct gactacaact ataagctgcc tgatgatttt acaggctgcg
     3181 tgatcgcctg gaactctaac aacctggata gcaaggtggg cggcaactac aactaccggt
     3241 accggctgtt tcgcaagtct aacctgaaac ctttcgagag agacatctcc acagagatct
     3301 accaggccgg ttctaagcct tgtaacgggg tggaaggctt caactgttac ttccctctgc
     3361 aaagctacgg cttccagcct accaatggag tcggctacca gccataccgg gtggtcgtgc
     3421 tgtccttcga gttactccac gcccccgcca ccgtctgcgg tggaggaggc ctggtgccac
     3481 agcagagcgg aggcctgaac gacatcttcg aggcccagaa gatcgagtgg cacgagggct
     3541 gatgaggatc cagatctgct gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3601 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3661 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3721 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3781 tgggtaccca ggtgctgaag aattgacccg gttcctcctg ggccagaaag aagcaggcac
     3841 atccccttct ctgtgacaca ccctgtccac gcccctggtt cttagttcca gccccactca
     3901 taggacactc atagctcagg agggctccgc cttcaatccc acccgctaaa gtacttggag
     3961 cggtctctcc ctccctcatc agcccaccaa accaaaccta gcctccaaga gtgggaagaa
     4021 attaaagcaa gataggctat taagtgcaga gggagagaaa atgcctccaa catgtgagga
     4081 agtaatgaga gaaatcatag aattttaagg ccatgattta aggccatcat ggccttaatc
     4141 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4201 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4261 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4741 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4861 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4921 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4981 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tcgggggggg
     5161 ggggcgctga ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc
     5221 cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga
     5281 ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg
     5341 cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt
     5401 caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac
     5461 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     5521 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     5581 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     5641 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     5701 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     5761 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     5821 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     5881 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     5941 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     6001 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     6061 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     6121 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     6181 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa
     6241 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     6301 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     6361 gacacaacgt ggctttcccc ccccccccat tattgaagca tttatcaggg ttattgtctc
     6421 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6481 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6541 aaaaataggc gtatcacg
//
