LOCUS       sequence-347376-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing L452R-E484Q 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-E484Q-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..44
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(82..104)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     204..223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        374..753
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        754..957
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     908..928
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     954..978
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2887..2910
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3532..3576
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3599..3823
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4190..4207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4361..4949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4441..4460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5491..6306)
                     /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(5491..6306)
                     /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     6214..6233
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6298..6317
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
       61 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      121 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      181 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      241 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagat tggctattgg
      301 ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca
      361 ttaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      421 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      481 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      541 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      601 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      661 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      721 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      781 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      841 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      901 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      961 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     1021 caccgggacc gatccagcct ccatcggctc gcatctctcc ttcacgcgcc cgccgcccta
     1081 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     1141 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     1201 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     1261 ttgctcaact ctagttaacg gtggagggca gtgtagtctg agcagtactc gttgctgccg
     1321 cgcgcgccac cagacataat agctgacaga ctaacagact gttcctttcc atgggtcttt
     1381 tctgcagtca ccgtcgtcga cacgtgtgat cagatatcgc ggccgctcta gaccaccatg
     1441 ggatggtcat gtatcatcct ttttctagta gcaactgcaa ccggtgtaca ttccgcacca
     1501 gagctgctgg gaggacctag cgtgttcctg tttcccccta agcctaagga caccctgatg
     1561 atcagccgga ccccagaggt gacatgcgtg gtggtggacg tgtcccacga ggaccccgag
     1621 gtgaagttca actggtacgt ggatggcgtg gaggtgcaca atgccaagac caagccaagg
     1681 gaggagcagt acaactctac ctatcgcgtg gtgagcgtgc tgacagtgct gcaccaggac
     1741 tggctgaacg gcaaggagta caagtgtaag gtgtctaata aggccctgcc tgccccaatc
     1801 gagaagacca tcagcaaggc aaagggacag ccaagggagc ctcaggtgta taccctgcca
     1861 cccagccgcg acgagctgac aaagaaccag gtgtccctgt actgcctggt gaagggcttc
     1921 tatccttccg atatcgccgt ggagtgggag tctaatggcc agccagagaa caattacaag
     1981 accacacctc cagtgctgga ctccgatggc tctttctttc tgtattccaa gctgaccgtg
     2041 gataagagcc ggtggcagca gggcaacgtg tttagctgtt ccgtgatgca cgaggccctg
     2101 cacaatcact acacacagaa gtctctgagc ctgtcccccg gcaagggagg aggaggcagc
     2161 ggcggaggag gctccggagg cggcggctct ggcggcggcg gcagcgcccc tgaactgctg
     2221 ggaggaccaa gcgtgttcct gtttccacct aagcccaaag ataccctgat gatctccagg
     2281 acccctgaag tcacctgcgt ggtcgtggac gtgtctcatg aggaccccga agtcaagttc
     2341 aactggtacg tggacggagt cgaagtccat aacgccaaga ccaagcctag agaagagcag
     2401 tacaactcca cctatagagt ggtgtctgtg ctgacagtgc tgcatcagga ttggctgaat
     2461 ggcaaggaat ataagtgtaa ggtgagcaat aaggccctgc ccgcccctat cgagaagacc
     2521 atctctaaag ccaaaggaca gccaagggag ccacaggtgt atacactgcc acccagccgg
     2581 gacgagctga ccaagaacca ggtgagcctg acatgtctgg tcaagggctt ctaccccagt
     2641 gacattgctg tggagtggga gtccaatggc cagcctgaga acaattataa aactacccca
     2701 ccagtgctgg acagcgatgg ctccttcttt ctgaccagca agctgacagt ggataagtcc
     2761 cgctggcagc agggaaacgt gttttcttgt agcgtgatgc atgaggccct gcataaccac
     2821 tacacccaga agtccctgtc tctgagccca ggcaagggag gcagcggagg cggaggcagc
     2881 ggaggcctgg aggtgctgtt tcagggccct ccaaacatta ccaacctctg ccccttcggc
     2941 gaggtgttta acgccacaag atttgcctcc gtttacgcct ggaatagaaa gagaatcagc
     3001 aattgtgtgg ccgactactc cgtgctgtat aacagcgcct ctttcagcac cttcaagtgc
     3061 tacggcgttt ccccaacaaa gctgaatgac ctgtgcttca ccaacgtgta cgccgactcc
     3121 ttcgtaatta gaggcgatga ggtgcggcag atcgcaccag gccagaccgg taagatcgct
     3181 gactacaact ataagctgcc tgatgatttt acaggctgcg tgatcgcctg gaactctaac
     3241 aacctggata gcaaggtggg cggcaactac aactaccggt accggctgtt tcgcaagtct
     3301 aacctgaaac ctttcgagag agacatctcc acagagatct accaggccgg ttctacacct
     3361 tgtaacgggg tgcagggctt caactgttac ttccctctgc aaagctacgg cttccagcct
     3421 accaatggag tcggctacca gccataccgg gtggtcgtgc tgtccttcga gttactccac
     3481 gcccccgcca ccgtctgcgg tggaggaggc ctggtgccac agcagagcgg aggcctgaac
     3541 gacatcttcg aggcccagaa gatcgagtgg cacgagggct gatgaggatc cagatctgct
     3601 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3661 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3721 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3781 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tgggtaccca ggtgctgaag
     3841 aattgacccg gttcctcctg ggccagaaag aagcaggcac atccccttct ctgtgacaca
     3901 ccctgtccac gcccctggtt cttagttcca gccccactca taggacactc atagctcagg
     3961 agggctccgc cttcaatccc acccgctaaa gtacttggag cggtctctcc ctccctcatc
     4021 agcccaccaa accaaaccta gcctccaaga gtgggaagaa attaaagcaa gataggctat
     4081 taagtgcaga gggagagaaa atgcctccaa catgtgagga agtaatgaga gaaatcatag
     4141 aattttaagg ccatgattta aggccatcat ggccttaatc ttccgcttcc tcgctcactg
     4201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4801 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5161 gtctatttcg ttcatccata gttgcctgac tcgggggggg ggggcgctga ggtctgcctc
     5221 gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc agccagaaag
     5281 tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg attttgaact
     5341 tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact
     5401 cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg taatgctctg
     5461 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     5521 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     5581 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     5641 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     5701 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     5761 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     5821 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     5881 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     5941 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     6001 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     6061 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     6121 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     6181 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     6241 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg
     6301 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     6361 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttcccc
     6421 ccccccccat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6481 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6541 tgacgtctaa gaaaccat
//
