LOCUS       sequence-347369-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing K417N-E484K-N501Y 
            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-E484K-N501Y-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..20
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(58..80)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     180..199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        350..729
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        730..933
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     884..904
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     930..954
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2863..2886
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3508..3552
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3575..3799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4166..4183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4337..4925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4417..4436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5467..6282)
                     /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(5467..6282)
                     /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     6190..6209
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6274..6293
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
       61 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      241 cgtaaggaga aaataccgca tcagattggc tattggccat tgcatacgtt gtatccatat
      301 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
      361 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      421 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      481 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      541 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      601 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      661 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      721 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      781 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      841 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      901 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      961 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccat
     1021 cggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     1081 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     1141 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     1201 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctag ttaacggtgg
     1261 agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga cataatagct
     1321 gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt cgtcgacacg
     1381 tgtgatcaga tatcgcggcc gctctagacc accatgggat ggtcatgtat catccttttt
     1441 ctagtagcaa ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg acctagcgtg
     1501 ttcctgtttc cccctaagcc taaggacacc ctgatgatca gccggacccc agaggtgaca
     1561 tgcgtggtgg tggacgtgtc ccacgaggac cccgaggtga agttcaactg gtacgtggat
     1621 ggcgtggagg tgcacaatgc caagaccaag ccaagggagg agcagtacaa ctctacctat
     1681 cgcgtggtga gcgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtacaag
     1741 tgtaaggtgt ctaataaggc cctgcctgcc ccaatcgaga agaccatcag caaggcaaag
     1801 ggacagccaa gggagcctca ggtgtatacc ctgccaccca gccgcgacga gctgacaaag
     1861 aaccaggtgt ccctgtactg cctggtgaag ggcttctatc cttccgatat cgccgtggag
     1921 tgggagtcta atggccagcc agagaacaat tacaagacca cacctccagt gctggactcc
     1981 gatggctctt tctttctgta ttccaagctg accgtggata agagccggtg gcagcagggc
     2041 aacgtgttta gctgttccgt gatgcacgag gccctgcaca atcactacac acagaagtct
     2101 ctgagcctgt cccccggcaa gggaggagga ggcagcggcg gaggaggctc cggaggcggc
     2161 ggctctggcg gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt gttcctgttt
     2221 ccacctaagc ccaaagatac cctgatgatc tccaggaccc ctgaagtcac ctgcgtggtc
     2281 gtggacgtgt ctcatgagga ccccgaagtc aagttcaact ggtacgtgga cggagtcgaa
     2341 gtccataacg ccaagaccaa gcctagagaa gagcagtaca actccaccta tagagtggtg
     2401 tctgtgctga cagtgctgca tcaggattgg ctgaatggca aggaatataa gtgtaaggtg
     2461 agcaataagg ccctgcccgc ccctatcgag aagaccatct ctaaagccaa aggacagcca
     2521 agggagccac aggtgtatac actgccaccc agccgggacg agctgaccaa gaaccaggtg
     2581 agcctgacat gtctggtcaa gggcttctac cccagtgaca ttgctgtgga gtgggagtcc
     2641 aatggccagc ctgagaacaa ttataaaact accccaccag tgctggacag cgatggctcc
     2701 ttctttctga ccagcaagct gacagtggat aagtcccgct ggcagcaggg aaacgtgttt
     2761 tcttgtagcg tgatgcatga ggccctgcat aaccactaca cccagaagtc cctgtctctg
     2821 agcccaggca agggaggcag cggaggcgga ggcagcggag gcctggaggt gctgtttcag
     2881 ggccctccaa acattaccaa cctctgcccc ttcggcgagg tgtttaacgc cacaagattt
     2941 gcctccgttt acgcctggaa tagaaagaga atcagcaatt gtgtggccga ctactccgtg
     3001 ctgtataaca gcgcctcttt cagcaccttc aagtgctacg gcgtttcccc aacaaagctg
     3061 aatgacctgt gcttcaccaa cgtgtacgcc gactccttcg taattagagg cgatgaggtg
     3121 cggcagatcg caccaggcca gaccggtaac atcgctgact acaactataa gctgcctgat
     3181 gattttacag gctgcgtgat cgcctggaac tctaacaacc tggatagcaa ggtgggcggc
     3241 aactacaact acctgtaccg gctgtttcgc aagtctaacc tgaaaccttt cgagagagac
     3301 atctccacag agatctacca ggccggttct acaccttgta acggggtgaa aggcttcaac
     3361 tgttacttcc ctctgcaaag ctacggcttc cagcctacct acggagtcgg ctaccagcca
     3421 taccgggtgg tcgtgctgtc cttcgagtta ctccacgccc ccgccaccgt ctgcggtgga
     3481 ggaggcctgg tgccacagca gagcggaggc ctgaacgaca tcttcgaggc ccagaagatc
     3541 gagtggcacg agggctgatg aggatccaga tctgctgtgc cttctagttg ccagccatct
     3601 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3661 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3721 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3781 gatgcggtgg gctctatggg tacccaggtg ctgaagaatt gacccggttc ctcctgggcc
     3841 agaaagaagc aggcacatcc ccttctctgt gacacaccct gtccacgccc ctggttctta
     3901 gttccagccc cactcatagg acactcatag ctcaggaggg ctccgccttc aatcccaccc
     3961 gctaaagtac ttggagcggt ctctccctcc ctcatcagcc caccaaacca aacctagcct
     4021 ccaagagtgg gaagaaatta aagcaagata ggctattaag tgcagaggga gagaaaatgc
     4081 ctccaacatg tgaggaagta atgagagaaa tcatagaatt ttaaggccat gatttaaggc
     4141 catcatggcc ttaatcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4741 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5161 cctgactcgg gggggggggg cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat
     5221 accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg ttgatgagag
     5281 ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg gaacggtctg
     5341 cgttgtcggg aagatgcgtg atctgatcct tcaactcagc aaaagttcga tttattcaac
     5401 aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa
     5461 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     5521 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     5581 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     5641 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     5701 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     5761 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     5821 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     5881 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     5941 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     6001 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     6061 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     6121 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     6181 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     6241 tcgcggcctc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     6301 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     6361 acatcagaga ttttgagaca caacgtggct ttcccccccc ccccattatt gaagcattta
     6421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6481 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6541 catgacatta acctataa
//
