LOCUS       sequence-347374-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain containing K417T-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-K417T-E484K-N501Y-...
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..57
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(95..117)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     217..236
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        387..766
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        767..970
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     921..941
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     967..991
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2900..2923
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3773..3817
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3840..4064
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4431..4448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4602..5190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4682..4701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5732..6547)
                     /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(5732..6547)
                     /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     6455..6474
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6539..6558
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
       61 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
      121 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      181 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
      241 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      301 gattggctat tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
      361 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
      421 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      481 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      541 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      601 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      661 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      721 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      781 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      841 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      901 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      961 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     1021 cctccataga agacaccggg accgatccag cctccatcgg ctcgcatctc tccttcacgc
     1081 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     1141 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     1201 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     1261 tgcctgaccc tgcttgctca actctagtta acggtggagg gcagtgtagt ctgagcagta
     1321 ctcgttgctg ccgcgcgcgc caccagacat aatagctgac agactaacag actgttcctt
     1381 tccatgggtc ttttctgcag tcaccgtcgt cgacacgtgt gatcagatat cgcggccgct
     1441 ctagaccacc atgggatggt catgtatcat cctttttcta gtagcaactg caaccggtgt
     1501 acattccgca ccagagctgc tgggaggacc tagcgtgttc ctgtttcccc ctaagcctaa
     1561 ggacaccctg atgatcagcc ggaccccaga ggtgacatgc gtggtggtgg acgtgtccca
     1621 cgaggacccc gaggtgaagt tcaactggta cgtggatggc gtggaggtgc acaatgccaa
     1681 gaccaagcca agggaggagc agtacaactc tacctatcgc gtggtgagcg tgctgacagt
     1741 gctgcaccag gactggctga acggcaagga gtacaagtgt aaggtgtcta ataaggccct
     1801 gcctgcccca atcgagaaga ccatcagcaa ggcaaaggga cagccaaggg agcctcaggt
     1861 gtataccctg ccacccagcc gcgacgagct gacaaagaac caggtgtccc tgtactgcct
     1921 ggtgaagggc ttctatcctt ccgatatcgc cgtggagtgg gagtctaatg gccagccaga
     1981 gaacaattac aagaccacac ctccagtgct ggactccgat ggctctttct ttctgtattc
     2041 caagctgacc gtggataaga gccggtggca gcagggcaac gtgtttagct gttccgtgat
     2101 gcacgaggcc ctgcacaatc actacacaca gaagtctctg agcctgtccc ccggcaaggg
     2161 aggaggaggc agcggcggag gaggctccgg aggcggcggc tctggcggcg gcggcagcgc
     2221 ccctgaactg ctgggaggac caagcgtgtt cctgtttcca cctaagccca aagataccct
     2281 gatgatctcc aggacccctg aagtcacctg cgtggtcgtg gacgtgtctc atgaggaccc
     2341 cgaagtcaag ttcaactggt acgtggacgg agtcgaagtc cataacgcca agaccaagcc
     2401 tagagaagag cagtacaact ccacctatag agtggtgtct gtgctgacag tgctgcatca
     2461 ggattggctg aatggcaagg aatataagtg taaggtgagc aataaggccc tgcccgcccc
     2521 tatcgagaag accatctcta aagccaaagg acagccaagg gagccacagg tgtatacact
     2581 gccacccagc cgggacgagc tgaccaagaa ccaggtgagc ctgacatgtc tggtcaaggg
     2641 cttctacccc agtgacattg ctgtggagtg ggagtccaat ggccagcctg agaacaatta
     2701 taaaactacc ccaccagtgc tggacagcga tggctccttc tttctgacca gcaagctgac
     2761 agtggataag tcccgctggc agcagggaaa cgtgttttct tgtagcgtga tgcatgaggc
     2821 cctgcataac cactacaccc agaagtccct gtctctgagc ccaggcaagg gaggcagcgg
     2881 aggcggaggc agcggaggcc tggaggtgct gtttcagggc cctcgggtgc agcctacaga
     2941 gagcatcgtg cgatttccaa acattaccaa cctctgcccc ttcggcgagg tgtttaacgc
     3001 cacaagattt gcctccgttt acgcctggaa tagaaagaga atcagcaatt gtgtggccga
     3061 ctactccgtg ctgtataaca gcgcctcttt cagcaccttc aagtgctacg gcgtttcccc
     3121 aacaaagctg aatgacctgt gcttcaccaa cgtgtacgcc gactccttcg taattagagg
     3181 cgatgaggtg cggcagatcg caccaggcca gaccggtacc atcgctgact acaactataa
     3241 gctgcctgat gattttacag gctgcgtgat cgcctggaac tctaacaacc tggatagcaa
     3301 ggtgggcggc aactacaact acctgtaccg gctgtttcgc aagtctaacc tgaaaccttt
     3361 cgagagagac atctccacag agatctacca ggccggttct acaccttgta acggggtgaa
     3421 aggcttcaac tgttacttcc ctctgcaaag ctacggcttc cagcctacct acggagtcgg
     3481 ctaccagcca taccgggtgg tcgtgctgtc cttcgagtta ctccacgccc ccgccaccgt
     3541 ctgcggtcct aagaagtcca ccaatctggt taagaacaaa tgcgtgaact tcaacttcaa
     3601 cggcctgacc gggaccggcg tgctgaccga aagcaacaaa aagttcctcc ccttccagca
     3661 gttcggccgt gatatcgctg acaccacaga tgccgtcaga gatccacaga ccctggaaat
     3721 cctggatatt acaccctgct ccggaggagg cctggtgcca cagcagagcg gaggcctgaa
     3781 cgacatcttc gaggcccaga agatcgagtg gcacgagggc tgatgaggat ccagatctgc
     3841 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3901 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3961 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4021 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atgggtaccc aggtgctgaa
     4081 gaattgaccc ggttcctcct gggccagaaa gaagcaggca catccccttc tctgtgacac
     4141 accctgtcca cgcccctggt tcttagttcc agccccactc ataggacact catagctcag
     4201 gagggctccg ccttcaatcc cacccgctaa agtacttgga gcggtctctc cctccctcat
     4261 cagcccacca aaccaaacct agcctccaag agtgggaaga aattaaagca agataggcta
     4321 ttaagtgcag agggagagaa aatgcctcca acatgtgagg aagtaatgag agaaatcata
     4381 gaattttaag gccatgattt aaggccatca tggccttaat cttccgcttc ctcgctcact
     4441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5401 tgtctatttc gttcatccat agttgcctga ctcggggggg gggggcgctg aggtctgcct
     5461 cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa
     5521 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac
     5581 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
     5641 tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct
     5701 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     5761 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     5821 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     5881 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     5941 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     6001 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     6061 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     6121 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     6181 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     6241 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     6301 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     6361 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     6421 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     6481 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
     6541 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     6601 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc
     6661 ccccccccca ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6781 ctgacg
//
