LOCUS       sequence-347372-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 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 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-K417N-E484K-N501Y-AVI
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(134..949)
                     /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(134..949)
                     /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     857..876
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     941..960
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1227..1245
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1283..1305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1405..1424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1575..1954
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1955..2158
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2109..2129
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2155..2179
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4088..4111
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4961..5005
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5028..5252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5619..5636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5790..6378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5870..5889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt
       61 attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat
      121 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
      181 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
      241 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
      301 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
      361 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
      421 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
      481 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
      541 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
      601 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
      661 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
      721 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
      781 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
      841 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
      901 aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg
      961 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     1021 caatgtaaca tcagagattt tgagacacaa cgtggctttc cccccccccc cattattgaa
     1081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1201 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1261 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1321 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     1381 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     1441 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcaga ttggctattg
     1501 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     1561 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     1621 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1681 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1741 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1801 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1861 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1921 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1981 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2041 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2101 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     2161 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     2221 acaccgggac cgatccagcc tccatcggct cgcatctctc cttcacgcgc ccgccgccct
     2281 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     2341 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     2401 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     2461 cttgctcaac tctagttaac ggtggagggc agtgtagtct gagcagtact cgttgctgcc
     2521 gcgcgcgcca ccagacataa tagctgacag actaacagac tgttcctttc catgggtctt
     2581 ttctgcagtc accgtcgtcg acacgtgtga tcagatatcg cggccgctct agaccaccat
     2641 gggatggtca tgtatcatcc tttttctagt agcaactgca accggtgtac attccgcacc
     2701 agagctgctg ggaggaccta gcgtgttcct gtttccccct aagcctaagg acaccctgat
     2761 gatcagccgg accccagagg tgacatgcgt ggtggtggac gtgtcccacg aggaccccga
     2821 ggtgaagttc aactggtacg tggatggcgt ggaggtgcac aatgccaaga ccaagccaag
     2881 ggaggagcag tacaactcta cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga
     2941 ctggctgaac ggcaaggagt acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat
     3001 cgagaagacc atcagcaagg caaagggaca gccaagggag cctcaggtgt ataccctgcc
     3061 acccagccgc gacgagctga caaagaacca ggtgtccctg tactgcctgg tgaagggctt
     3121 ctatccttcc gatatcgccg tggagtggga gtctaatggc cagccagaga acaattacaa
     3181 gaccacacct ccagtgctgg actccgatgg ctctttcttt ctgtattcca agctgaccgt
     3241 ggataagagc cggtggcagc agggcaacgt gtttagctgt tccgtgatgc acgaggccct
     3301 gcacaatcac tacacacaga agtctctgag cctgtccccc ggcaagggag gaggaggcag
     3361 cggcggagga ggctccggag gcggcggctc tggcggcggc ggcagcgccc ctgaactgct
     3421 gggaggacca agcgtgttcc tgtttccacc taagcccaaa gataccctga tgatctccag
     3481 gacccctgaa gtcacctgcg tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt
     3541 caactggtac gtggacggag tcgaagtcca taacgccaag accaagccta gagaagagca
     3601 gtacaactcc acctatagag tggtgtctgt gctgacagtg ctgcatcagg attggctgaa
     3661 tggcaaggaa tataagtgta aggtgagcaa taaggccctg cccgccccta tcgagaagac
     3721 catctctaaa gccaaaggac agccaaggga gccacaggtg tatacactgc cacccagccg
     3781 ggacgagctg accaagaacc aggtgagcct gacatgtctg gtcaagggct tctaccccag
     3841 tgacattgct gtggagtggg agtccaatgg ccagcctgag aacaattata aaactacccc
     3901 accagtgctg gacagcgatg gctccttctt tctgaccagc aagctgacag tggataagtc
     3961 ccgctggcag cagggaaacg tgttttcttg tagcgtgatg catgaggccc tgcataacca
     4021 ctacacccag aagtccctgt ctctgagccc aggcaaggga ggcagcggag gcggaggcag
     4081 cggaggcctg gaggtgctgt ttcagggccc tcgggtgcag cctacagaga gcatcgtgcg
     4141 atttccaaac attaccaacc tctgcccctt cggcgaggtg tttaacgcca caagatttgc
     4201 ctccgtttac gcctggaata gaaagagaat cagcaattgt gtggccgact actccgtgct
     4261 gtataacagc gcctctttca gcaccttcaa gtgctacggc gtttccccaa caaagctgaa
     4321 tgacctgtgc ttcaccaacg tgtacgccga ctccttcgta attagaggcg atgaggtgcg
     4381 gcagatcgca ccaggccaga ccggtaacat cgctgactac aactataagc tgcctgatga
     4441 ttttacaggc tgcgtgatcg cctggaactc taacaacctg gatagcaagg tgggcggcaa
     4501 ctacaactac ctgtaccggc tgtttcgcaa gtctaacctg aaacctttcg agagagacat
     4561 ctccacagag atctaccagg ccggttctac accttgtaac ggggtgaaag gcttcaactg
     4621 ttacttccct ctgcaaagct acggcttcca gcctacctac ggagtcggct accagccata
     4681 ccgggtggtc gtgctgtcct tcgagttact ccacgccccc gccaccgtct gcggtcctaa
     4741 gaagtccacc aatctggtta agaacaaatg cgtgaacttc aacttcaacg gcctgaccgg
     4801 gaccggcgtg ctgaccgaaa gcaacaaaaa gttcctcccc ttccagcagt tcggccgtga
     4861 tatcgctgac accacagatg ccgtcagaga tccacagacc ctggaaatcc tggatattac
     4921 accctgctcc ggaggaggcc tggtgccaca gcagagcgga ggcctgaacg acatcttcga
     4981 ggcccagaag atcgagtggc acgagggctg atgaggatcc agatctgctg tgccttctag
     5041 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     5101 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     5161 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     5221 caggcatgct ggggatgcgg tgggctctat gggtacccag gtgctgaaga attgacccgg
     5281 ttcctcctgg gccagaaaga agcaggcaca tccccttctc tgtgacacac cctgtccacg
     5341 cccctggttc ttagttccag ccccactcat aggacactca tagctcagga gggctccgcc
     5401 ttcaatccca cccgctaaag tacttggagc ggtctctccc tccctcatca gcccaccaaa
     5461 ccaaacctag cctccaagag tgggaagaaa ttaaagcaag ataggctatt aagtgcagag
     5521 ggagagaaaa tgcctccaac atgtgaggaa gtaatgagag aaatcataga attttaaggc
     5581 catgatttaa ggccatcatg gccttaatct tccgcttcct cgctcactga ctcgctgcgc
     5641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6601 tcatccatag ttgcctgact cggggggggg gggcgctgag gtctgcctcg tgaagaaggt
     6661 gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca
     6721 cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc
     6781 acggaa
//
