LOCUS       sequence-347373-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain containing L452R-T478K 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-T478K-AVI
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        277..656
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        657..860
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     811..831
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     857..881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2790..2813
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3663..3707
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3730..3954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4321..4338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4492..5080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4572..4591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5622..6437)
                     /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(5622..6437)
                     /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     6345..6364
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6429..6448
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6715..6733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6771..6786,1..7))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
       61 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
      121 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
      181 taccgcatca gattggctat tggccattgc atacgttgta tccatatcat aatatgtaca
      241 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      301 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      361 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      421 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      481 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      541 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      601 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      661 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      721 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      781 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      841 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      901 gctgttttga cctccataga agacaccggg accgatccag cctccatcgg ctcgcatctc
      961 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     1021 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     1081 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     1141 ctccacgctt tgcctgaccc tgcttgctca actctagtta acggtggagg gcagtgtagt
     1201 ctgagcagta ctcgttgctg ccgcgcgcgc caccagacat aatagctgac agactaacag
     1261 actgttcctt tccatgggtc ttttctgcag tcaccgtcgt cgacacgtgt gatcagatat
     1321 cgcggccgct ctagaccacc atgggatggt catgtatcat cctttttcta gtagcaactg
     1381 caaccggtgt acattccgca ccagagctgc tgggaggacc tagcgtgttc ctgtttcccc
     1441 ctaagcctaa ggacaccctg atgatcagcc ggaccccaga ggtgacatgc gtggtggtgg
     1501 acgtgtccca cgaggacccc gaggtgaagt tcaactggta cgtggatggc gtggaggtgc
     1561 acaatgccaa gaccaagcca agggaggagc agtacaactc tacctatcgc gtggtgagcg
     1621 tgctgacagt gctgcaccag gactggctga acggcaagga gtacaagtgt aaggtgtcta
     1681 ataaggccct gcctgcccca atcgagaaga ccatcagcaa ggcaaaggga cagccaaggg
     1741 agcctcaggt gtataccctg ccacccagcc gcgacgagct gacaaagaac caggtgtccc
     1801 tgtactgcct ggtgaagggc ttctatcctt ccgatatcgc cgtggagtgg gagtctaatg
     1861 gccagccaga gaacaattac aagaccacac ctccagtgct ggactccgat ggctctttct
     1921 ttctgtattc caagctgacc gtggataaga gccggtggca gcagggcaac gtgtttagct
     1981 gttccgtgat gcacgaggcc ctgcacaatc actacacaca gaagtctctg agcctgtccc
     2041 ccggcaaggg aggaggaggc agcggcggag gaggctccgg aggcggcggc tctggcggcg
     2101 gcggcagcgc ccctgaactg ctgggaggac caagcgtgtt cctgtttcca cctaagccca
     2161 aagataccct gatgatctcc aggacccctg aagtcacctg cgtggtcgtg gacgtgtctc
     2221 atgaggaccc cgaagtcaag ttcaactggt acgtggacgg agtcgaagtc cataacgcca
     2281 agaccaagcc tagagaagag cagtacaact ccacctatag agtggtgtct gtgctgacag
     2341 tgctgcatca ggattggctg aatggcaagg aatataagtg taaggtgagc aataaggccc
     2401 tgcccgcccc tatcgagaag accatctcta aagccaaagg acagccaagg gagccacagg
     2461 tgtatacact gccacccagc cgggacgagc tgaccaagaa ccaggtgagc ctgacatgtc
     2521 tggtcaaggg cttctacccc agtgacattg ctgtggagtg ggagtccaat ggccagcctg
     2581 agaacaatta taaaactacc ccaccagtgc tggacagcga tggctccttc tttctgacca
     2641 gcaagctgac agtggataag tcccgctggc agcagggaaa cgtgttttct tgtagcgtga
     2701 tgcatgaggc cctgcataac cactacaccc agaagtccct gtctctgagc ccaggcaagg
     2761 gaggcagcgg aggcggaggc agcggaggcc tggaggtgct gtttcagggc cctcgggtgc
     2821 agcctacaga gagcatcgtg cgatttccaa acattaccaa cctctgcccc ttcggcgagg
     2881 tgtttaacgc cacaagattt gcctccgttt acgcctggaa tagaaagaga atcagcaatt
     2941 gtgtggccga ctactccgtg ctgtataaca gcgcctcttt cagcaccttc aagtgctacg
     3001 gcgtttcccc aacaaagctg aatgacctgt gcttcaccaa cgtgtacgcc gactccttcg
     3061 taattagagg cgatgaggtg cggcagatcg caccaggcca gaccggtaag atcgctgact
     3121 acaactataa gctgcctgat gattttacag gctgcgtgat cgcctggaac tctaacaacc
     3181 tggatagcaa ggtgggcggc aactacaact acagataccg gctgtttcgc aagtctaacc
     3241 tgaaaccttt cgagagagac atctccacag agatctacca ggccggttct aagccttgta
     3301 acggggtgga aggcttcaac tgttacttcc ctctgcaaag ctacggcttc cagcctacca
     3361 atggagtcgg ctaccagcca taccgggtgg tcgtgctgtc cttcgagtta ctccacgccc
     3421 ccgccaccgt ctgcggtcct aagaagtcca ccaatctggt taagaacaaa tgcgtgaact
     3481 tcaacttcaa cggcctgact gggaccggcg tgctgaccga aagcaacaaa aagttcctcc
     3541 ccttccagca gttcggccgt gatatcgctg acaccacaga tgccgtcaga gatccacaga
     3601 ccctggaaat cctggatatt acaccctgct ccggaggagg cctggtgcca cagcagagcg
     3661 gaggcctgaa cgacatcttc gaggcccaga agatcgagtg gcacgagggc tgatgaggat
     3721 ccagatctgc tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3781 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3841 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atgggtaccc
     3961 aggtgctgaa gaattgaccc ggttcctcct gggccagaaa gaagcaggca catccccttc
     4021 tctgtgacac accctgtcca cgcccctggt tcttagttcc agccccactc ataggacact
     4081 catagctcag gagggctccg ccttcaatcc cacccgctaa agtacttgga gcggtctctc
     4141 cctccctcat cagcccacca aaccaaacct agcctccaag agtgggaaga aattaaagca
     4201 agataggcta ttaagtgcag agggagagaa aatgcctcca acatgtgagg aagtaatgag
     4261 agaaatcata gaattttaag gccatgattt aaggccatca tggccttaat cttccgcttc
     4321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctcggggggg gggggcgctg
     5341 aggtctgcct cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc
     5401 cagccagaaa gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt
     5461 gattttgaac ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg
     5521 atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc
     5581 gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
     5641 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     5701 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     5761 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     5821 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     5881 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     5941 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     6001 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     6061 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     6121 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     6181 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     6241 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     6301 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     6361 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc
     6421 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     6481 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     6541 tggctttccc ccccccccca ttattgaagc atttatcagg gttattgtct catgagcgga
     6601 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6661 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6721 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6781 atgcag
//
