LOCUS       sequence-347366-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 domain containing L452R 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-L452R-AVI
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..144
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(182..204)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     304..323
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        474..853
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        854..1057
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1008..1028
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1054..1078
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2987..3010
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3860..3904
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3927..4151
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4518..4535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4689..5277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4769..4788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5819..6634)
                     /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(5819..6634)
                     /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     6542..6561
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6626..6645
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
       61 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
      121 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
      181 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
      241 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
      301 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
      361 gatgcgtaag gagaaaatac cgcatcagat tggctattgg ccattgcata cgttgtatcc
      421 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg
      481 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      541 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      601 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      661 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      721 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      781 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      841 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      901 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      961 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1021 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     1081 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     1141 ccatcggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     1201 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     1261 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     1321 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctagttaacg
     1381 gtggagggca gtgtagtctg agcagtactc gttgctgccg cgcgcgccac cagacataat
     1441 agctgacaga ctaacagact gttcctttcc atgggtcttt tctgcagtca ccgtcgtcga
     1501 cacgtgtgat cagatatcgc ggccgctcta gaccaccatg ggatggtcat gtatcatcct
     1561 ttttctagta gcaactgcaa ccggtgtaca ttccgcacca gagctgctgg gaggacctag
     1621 cgtgttcctg tttcccccta agcctaagga caccctgatg atcagccgga ccccagaggt
     1681 gacatgcgtg gtggtggacg tgtcccacga ggaccccgag gtgaagttca actggtacgt
     1741 ggatggcgtg gaggtgcaca atgccaagac caagccaagg gaggagcagt acaactctac
     1801 ctatcgcgtg gtgagcgtgc tgacagtgct gcaccaggac tggctgaacg gcaaggagta
     1861 caagtgtaag gtgtctaata aggccctgcc tgccccaatc gagaagacca tcagcaaggc
     1921 aaagggacag ccaagggagc ctcaggtgta taccctgcca cccagccgcg acgagctgac
     1981 aaagaaccag gtgtccctgt actgcctggt gaagggcttc tatccttccg atatcgccgt
     2041 ggagtgggag tctaatggcc agccagagaa caattacaag accacacctc cagtgctgga
     2101 ctccgatggc tctttctttc tgtattccaa gctgaccgtg gataagagcc ggtggcagca
     2161 gggcaacgtg tttagctgtt ccgtgatgca cgaggccctg cacaatcact acacacagaa
     2221 gtctctgagc ctgtcccccg gcaagggagg aggaggcagc ggcggaggag gctccggagg
     2281 cggcggctct ggcggcggcg gcagcgcccc tgaactgctg ggaggaccaa gcgtgttcct
     2341 gtttccacct aagcccaaag ataccctgat gatctccagg acccctgaag tcacctgcgt
     2401 ggtcgtggac gtgtctcatg aggaccccga agtcaagttc aactggtacg tggacggagt
     2461 cgaagtccat aacgccaaga ccaagcctag agaagagcag tacaactcca cctatagagt
     2521 ggtgtctgtg ctgacagtgc tgcatcagga ttggctgaat ggcaaggaat ataagtgtaa
     2581 ggtgagcaat aaggccctgc ccgcccctat cgagaagacc atctctaaag ccaaaggaca
     2641 gccaagggag ccacaggtgt atacactgcc acccagccgg gacgagctga ccaagaacca
     2701 ggtgagcctg acatgtctgg tcaagggctt ctaccccagt gacattgctg tggagtggga
     2761 gtccaatggc cagcctgaga acaattataa aactacccca ccagtgctgg acagcgatgg
     2821 ctccttcttt ctgaccagca agctgacagt ggataagtcc cgctggcagc agggaaacgt
     2881 gttttcttgt agcgtgatgc atgaggccct gcataaccac tacacccaga agtccctgtc
     2941 tctgagccca ggcaagggag gcagcggagg cggaggcagc ggaggcctgg aggtgctgtt
     3001 tcagggccct cgggtgcagc ctacagagag catcgtgcga tttccaaaca ttaccaacct
     3061 ctgccccttc ggcgaggtgt ttaacgccac aagatttgcc tccgtttacg cctggaatag
     3121 aaagagaatc agcaattgtg tggccgacta ctccgtgctg tataacagcg cctctttcag
     3181 caccttcaag tgctacggcg tttccccaac aaagctgaat gacctgtgct tcaccaacgt
     3241 gtacgccgac tccttcgtaa ttagaggcga tgaggtgcgg cagatcgcac caggccagac
     3301 cggtaagatc gctgactaca actataagct gcctgatgat tttacaggct gcgtgatcgc
     3361 ctggaactct aacaacctgg atagcaaggt gggcggcaac tacaactacc ggtaccggct
     3421 gtttcgcaag tctaacctga aacctttcga gagagacatc tccacagaga tctaccaggc
     3481 cggttctaca ccttgtaacg gggtggaagg cttcaactgt tacttccctc tgcaaagcta
     3541 cggcttccag cctaccaatg gagtcggcta ccagccatac cgggtggtcg tgctgtcctt
     3601 cgagttactc cacgcccccg ccaccgtctg cggtcctaag aagtccacca atctggttaa
     3661 gaacaaatgc gtgaacttca acttcaacgg cctgaccggg accggcgtgc tgaccgaaag
     3721 caacaaaaag ttcctcccct tccagcagtt cggccgtgat atcgctgaca ccacagatgc
     3781 cgtcagagat ccacagaccc tggaaatcct ggatattaca ccctgctccg gaggaggcct
     3841 ggtgccacag cagagcggag gcctgaacga catcttcgag gcccagaaga tcgagtggca
     3901 cgagggctga tgaggatcca gatctgctgt gccttctagt tgccagccat ctgttgtttg
     3961 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4021 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4081 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4141 gggctctatg ggtacccagg tgctgaagaa ttgacccggt tcctcctggg ccagaaagaa
     4201 gcaggcacat ccccttctct gtgacacacc ctgtccacgc ccctggttct tagttccagc
     4261 cccactcata ggacactcat agctcaggag ggctccgcct tcaatcccac ccgctaaagt
     4321 acttggagcg gtctctccct ccctcatcag cccaccaaac caaacctagc ctccaagagt
     4381 gggaagaaat taaagcaaga taggctatta agtgcagagg gagagaaaat gcctccaaca
     4441 tgtgaggaag taatgagaga aatcatagaa ttttaaggcc atgatttaag gccatcatgg
     4501 ccttaatctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5521 gggggggggg ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc
     5581 tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg
     5641 taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg
     5701 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
     5761 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     5821 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     5881 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     5941 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     6001 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     6061 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     6121 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     6181 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     6241 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     6301 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     6361 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     6421 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     6481 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     6541 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     6601 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     6661 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     6721 gattttgaga cacaacgtgg ctttcccccc ccccccatta ttgaagcatt tatcagggtt
     6781 attgtc
//
