LOCUS       sequence-347377-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain containing E484K mutation 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-E484K-AVI
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(118..933)
                     /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(118..933)
                     /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     841..860
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     925..944
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1211..1229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1267..1289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1389..1408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1559..1938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1939..2142
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2093..2113
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2139..2163
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4072..4095
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4945..4989
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5012..5236
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5603..5620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5774..6362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5854..5873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa caaagccgcc
       61 gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta
      121 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
      181 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
      241 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
      301 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
      361 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
      421 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
      481 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
      541 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
      601 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
      661 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
      721 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
      781 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
      841 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
      901 cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
      961 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     1021 attttgagac acaacgtggc tttccccccc cccccattat tgaagcattt atcagggtta
     1081 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1141 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1201 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     1261 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1321 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     1381 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     1441 gcacagatgc gtaaggagaa aataccgcat cagattggct attggccatt gcatacgttg
     1501 tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc gccatgttga
     1561 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1621 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1681 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1741 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1801 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1861 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1921 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1981 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2041 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2101 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     2161 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     2221 agcctccatc ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     2281 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     2341 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     2401 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctagt
     2461 taacggtgga gggcagtgta gtctgagcag tactcgttgc tgccgcgcgc gccaccagac
     2521 ataatagctg acagactaac agactgttcc tttccatggg tcttttctgc agtcaccgtc
     2581 gtcgacacgt gtgatcagat atcgcggccg ctctagacca ccatgggatg gtcatgtatc
     2641 atcctttttc tagtagcaac tgcaaccggt gtacattccg caccagagct gctgggagga
     2701 cctagcgtgt tcctgtttcc ccctaagcct aaggacaccc tgatgatcag ccggacccca
     2761 gaggtgacat gcgtggtggt ggacgtgtcc cacgaggacc ccgaggtgaa gttcaactgg
     2821 tacgtggatg gcgtggaggt gcacaatgcc aagaccaagc caagggagga gcagtacaac
     2881 tctacctatc gcgtggtgag cgtgctgaca gtgctgcacc aggactggct gaacggcaag
     2941 gagtacaagt gtaaggtgtc taataaggcc ctgcctgccc caatcgagaa gaccatcagc
     3001 aaggcaaagg gacagccaag ggagcctcag gtgtataccc tgccacccag ccgcgacgag
     3061 ctgacaaaga accaggtgtc cctgtactgc ctggtgaagg gcttctatcc ttccgatatc
     3121 gccgtggagt gggagtctaa tggccagcca gagaacaatt acaagaccac acctccagtg
     3181 ctggactccg atggctcttt ctttctgtat tccaagctga ccgtggataa gagccggtgg
     3241 cagcagggca acgtgtttag ctgttccgtg atgcacgagg ccctgcacaa tcactacaca
     3301 cagaagtctc tgagcctgtc ccccggcaag ggaggaggag gcagcggcgg aggaggctcc
     3361 ggaggcggcg gctctggcgg cggcggcagc gcccctgaac tgctgggagg accaagcgtg
     3421 ttcctgtttc cacctaagcc caaagatacc ctgatgatct ccaggacccc tgaagtcacc
     3481 tgcgtggtcg tggacgtgtc tcatgaggac cccgaagtca agttcaactg gtacgtggac
     3541 ggagtcgaag tccataacgc caagaccaag cctagagaag agcagtacaa ctccacctat
     3601 agagtggtgt ctgtgctgac agtgctgcat caggattggc tgaatggcaa ggaatataag
     3661 tgtaaggtga gcaataaggc cctgcccgcc cctatcgaga agaccatctc taaagccaaa
     3721 ggacagccaa gggagccaca ggtgtataca ctgccaccca gccgggacga gctgaccaag
     3781 aaccaggtga gcctgacatg tctggtcaag ggcttctacc ccagtgacat tgctgtggag
     3841 tgggagtcca atggccagcc tgagaacaat tataaaacta ccccaccagt gctggacagc
     3901 gatggctcct tctttctgac cagcaagctg acagtggata agtcccgctg gcagcaggga
     3961 aacgtgtttt cttgtagcgt gatgcatgag gccctgcata accactacac ccagaagtcc
     4021 ctgtctctga gcccaggcaa gggaggcagc ggaggcggag gcagcggagg cctggaggtg
     4081 ctgtttcagg gccctcgggt gcagcctaca gagagcatcg tgcgatttcc aaacattacc
     4141 aacctctgcc ccttcggcga ggtgtttaac gccacaagat ttgcctccgt ttacgcctgg
     4201 aatagaaaga gaatcagcaa ttgtgtggcc gactactccg tgctgtataa cagcgcctct
     4261 ttcagcacct tcaagtgcta cggcgtttcc ccaacaaagc tgaatgacct gtgcttcacc
     4321 aacgtgtacg ccgactcctt cgtaattaga ggcgatgagg tgcggcagat cgcaccaggc
     4381 cagaccggta agatcgctga ctacaactat aagctgcctg atgattttac aggctgcgtg
     4441 atcgcctgga actctaacaa cctggatagc aaggtgggcg gcaactacaa ctacctgtac
     4501 cggctgtttc gcaagtctaa cctgaaacct ttcgagagag acatctccac agagatctac
     4561 caggccggtt ctacaccttg taacggggtg aaaggcttca actgttactt ccctctgcaa
     4621 agctacggct tccagcctac caatggagtc ggctaccagc cataccgggt ggtcgtgctg
     4681 tccttcgagt tactccacgc ccccgccacc gtctgcggtc ctaagaagtc caccaatctg
     4741 gttaagaaca aatgcgtgaa cttcaacttc aacggcctga ccgggaccgg cgtgctgacc
     4801 gaaagcaaca aaaagttcct ccccttccag cagttcggcc gtgatatcgc tgacaccaca
     4861 gatgccgtca gagatccaca gaccctggaa atcctggata ttacaccctg ctccggagga
     4921 ggcctggtgc cacagcagag cggaggcctg aacgacatct tcgaggccca gaagatcgag
     4981 tggcacgagg gctgatgagg atccagatct gctgtgcctt ctagttgcca gccatctgtt
     5041 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5101 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5161 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     5221 gcggtgggct ctatgggtac ccaggtgctg aagaattgac ccggttcctc ctgggccaga
     5281 aagaagcagg cacatcccct tctctgtgac acaccctgtc cacgcccctg gttcttagtt
     5341 ccagccccac tcataggaca ctcatagctc aggagggctc cgccttcaat cccacccgct
     5401 aaagtacttg gagcggtctc tccctccctc atcagcccac caaaccaaac ctagcctcca
     5461 agagtgggaa gaaattaaag caagataggc tattaagtgc agagggagag aaaatgcctc
     5521 caacatgtga ggaagtaatg agagaaatca tagaatttta aggccatgat ttaaggccat
     5581 catggcctta atcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6601 gactcggggg gggggggcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc
     6661 aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt
     6721 tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt
     6781 tgtcgg
//
