LOCUS       sequence-347370-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 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 6786)
  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 6786)
  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-SD1-L452R-E484Q-AVI
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..42
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(80..102)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     202..221
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        372..751
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        752..955
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     906..926
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..976
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2885..2908
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3758..3802
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3825..4049
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4416..4433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4587..5175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4667..4686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5717..6532)
                     /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(5717..6532)
                     /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     6440..6459
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6524..6543
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
       61 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
      121 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      181 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
      241 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcagattg gctattggcc
      301 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
      361 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      421 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      481 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      541 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      601 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      661 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      721 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      781 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      841 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      901 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      961 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
     1021 ccgggaccga tccagcctcc atcggctcgc atctctcctt cacgcgcccg ccgccctacc
     1081 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     1141 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     1201 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     1261 gctcaactct agttaacggt ggagggcagt gtagtctgag cagtactcgt tgctgccgcg
     1321 cgcgccacca gacataatag ctgacagact aacagactgt tcctttccat gggtcttttc
     1381 tgcagtcacc gtcgtcgaca cgtgtgatca gatatcgcgg ccgctctaga ccaccatggg
     1441 atggtcatgt atcatccttt ttctagtagc aactgcaacc ggtgtacatt ccgcaccaga
     1501 gctgctggga ggacctagcg tgttcctgtt tccccctaag cctaaggaca ccctgatgat
     1561 cagccggacc ccagaggtga catgcgtggt ggtggacgtg tcccacgagg accccgaggt
     1621 gaagttcaac tggtacgtgg atggcgtgga ggtgcacaat gccaagacca agccaaggga
     1681 ggagcagtac aactctacct atcgcgtggt gagcgtgctg acagtgctgc accaggactg
     1741 gctgaacggc aaggagtaca agtgtaaggt gtctaataag gccctgcctg ccccaatcga
     1801 gaagaccatc agcaaggcaa agggacagcc aagggagcct caggtgtata ccctgccacc
     1861 cagccgcgac gagctgacaa agaaccaggt gtccctgtac tgcctggtga agggcttcta
     1921 tccttccgat atcgccgtgg agtgggagtc taatggccag ccagagaaca attacaagac
     1981 cacacctcca gtgctggact ccgatggctc tttctttctg tattccaagc tgaccgtgga
     2041 taagagccgg tggcagcagg gcaacgtgtt tagctgttcc gtgatgcacg aggccctgca
     2101 caatcactac acacagaagt ctctgagcct gtcccccggc aagggaggag gaggcagcgg
     2161 cggaggaggc tccggaggcg gcggctctgg cggcggcggc agcgcccctg aactgctggg
     2221 aggaccaagc gtgttcctgt ttccacctaa gcccaaagat accctgatga tctccaggac
     2281 ccctgaagtc acctgcgtgg tcgtggacgt gtctcatgag gaccccgaag tcaagttcaa
     2341 ctggtacgtg gacggagtcg aagtccataa cgccaagacc aagcctagag aagagcagta
     2401 caactccacc tatagagtgg tgtctgtgct gacagtgctg catcaggatt ggctgaatgg
     2461 caaggaatat aagtgtaagg tgagcaataa ggccctgccc gcccctatcg agaagaccat
     2521 ctctaaagcc aaaggacagc caagggagcc acaggtgtat acactgccac ccagccggga
     2581 cgagctgacc aagaaccagg tgagcctgac atgtctggtc aagggcttct accccagtga
     2641 cattgctgtg gagtgggagt ccaatggcca gcctgagaac aattataaaa ctaccccacc
     2701 agtgctggac agcgatggct ccttctttct gaccagcaag ctgacagtgg ataagtcccg
     2761 ctggcagcag ggaaacgtgt tttcttgtag cgtgatgcat gaggccctgc ataaccacta
     2821 cacccagaag tccctgtctc tgagcccagg caagggaggc agcggaggcg gaggcagcgg
     2881 aggcctggag gtgctgtttc agggccctcg ggtgcagcct acagagagca tcgtgcgatt
     2941 tccaaacatt accaacctct gccccttcgg cgaggtgttt aacgccacaa gatttgcctc
     3001 cgtttacgcc tggaatagaa agagaatcag caattgtgtg gccgactact ccgtgctgta
     3061 taacagcgcc tctttcagca ccttcaagtg ctacggcgtt tccccaacaa agctgaatga
     3121 cctgtgcttc accaacgtgt acgccgactc cttcgtaatt agaggcgatg aggtgcggca
     3181 gatcgcacca ggccagaccg gtaagatcgc tgactacaac tataagctgc ctgatgattt
     3241 tacaggctgc gtgatcgcct ggaactctaa caacctggat agcaaggtgg gcggcaacta
     3301 caactacaga taccggctgt ttcgcaagtc taacctgaaa cctttcgaga gagacatctc
     3361 cacagagatc taccaggccg gttctacacc ttgtaacggg gtgcagggct tcaactgtta
     3421 cttccctctg caaagctacg gcttccagcc taccaatgga gtcggctacc agccataccg
     3481 ggtggtcgtg ctgtccttcg agttactcca cgcccccgcc accgtctgcg gtcctaagaa
     3541 gtccaccaat ctggttaaga acaaatgcgt gaacttcaac ttcaacggcc tgaccgggac
     3601 cggcgtgctg accgaaagca acaaaaagtt cctccccttc cagcagttcg gccgtgatat
     3661 cgctgacacc acagatgccg tcagagatcc acagaccctg gaaatcctgg atattacacc
     3721 ctgctccgga ggaggcctgg tgccacagca gagcggaggc ctgaacgaca tcttcgaggc
     3781 ccagaagatc gagtggcacg agggctgatg aggatccaga tctgctgtgc cttctagttg
     3841 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3901 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3961 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4021 gcatgctggg gatgcggtgg gctctatggg tacccaggtg ctgaagaatt gacccggttc
     4081 ctcctgggcc agaaagaagc aggcacatcc ccttctctgt gacacaccct gtccacgccc
     4141 ctggttctta gttccagccc cactcatagg acactcatag ctcaggaggg ctccgccttc
     4201 aatcccaccc gctaaagtac ttggagcggt ctctccctcc ctcatcagcc caccaaacca
     4261 aacctagcct ccaagagtgg gaagaaatta aagcaagata ggctattaag tgcagaggga
     4321 gagaaaatgc ctccaacatg tgaggaagta atgagagaaa tcatagaatt ttaaggccat
     4381 gatttaaggc catcatggcc ttaatcttcc gcttcctcgc tcactgactc gctgcgctcg
     4441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5401 tccatagttg cctgactcgg gggggggggg cgctgaggtc tgcctcgtga agaaggtgtt
     5461 gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg
     5521 ttgatgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg
     5581 gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc aaaagttcga
     5641 tttattcaac aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc
     5701 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     5761 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     5821 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     5881 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     5941 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga
     6001 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     6061 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     6121 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     6181 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     6241 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     6301 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     6361 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     6421 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     6481 tggaatttaa tcgcggcctc gagcaagacg tttcccgttg aatatggctc ataacacccc
     6541 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     6601 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttcccccccc ccccattatt
     6661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6781 ccatta
//
