LOCUS       sequence-354500-        6543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain from the B.1.1.529 (Omicron) variant
            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 6543)
  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 6543)
  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(aa335-526)-B.1.1.529-AVI
FEATURES             Location/Qualifiers
     source          1..6543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        321..700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        701..904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     855..875
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     901..925
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2834..2857
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3464..3508
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3531..3755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4122..4139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4293..4881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4373..4392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5423..6238)
                     /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(5423..6238)
                     /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     6146..6165
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6230..6249
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6516..6534
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcagattgg
      241 ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
      301 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      721 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      781 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      841 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      901 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
      961 tagaagacac cgggaccgat ccagcctcca tcggctcgca tctctccttc acgcgcccgc
     1021 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     1081 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     1141 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     1201 accctgcttg ctcaactcta gttaacggtg gagggcagtg tagtctgagc agtactcgtt
     1261 gctgccgcgc gcgccaccag acataatagc tgacagacta acagactgtt cctttccatg
     1321 ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag atatcgcggc cgctctagac
     1381 caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg gtgtacattc
     1441 cgcacctgag ctgctgggcg gacctagtgt ctttctgttc ccccccaagc ctaaggacac
     1501 cctgatgatc agccggaccc ccgaggttac atgcgtggtg gtcgatgtgt cccacgagga
     1561 ccctgaagtg aagtttaatt ggtatgtgga cggcgtcgag gtccacaacg ccaagaccaa
     1621 gcctagagag gaacagtaca acagcaccta cagagtggtg agcgtgctga ccgtgctgca
     1681 ccaagattgg ctgaatggca aggagtacaa gtgcaaggtg agcaacaagg ccctgccagc
     1741 tcctattgag aaaaccatca gcaaagccaa gggccagcct cgggagcccc aggtgtatac
     1801 cctgcctcca tctcgcgacg agctgaccaa gaaccaggtg tctctgtact gcctggtcaa
     1861 gggcttttac cctagcgaca tcgccgtgga atgggaaagc aacggccagc ctgagaacaa
     1921 ctataaaaca acgccacctg tgctggactc tgatggcagc ttcttcctgt acagcaagct
     1981 gaccgtggac aagagcaggt ggcaacaggg caacgtgttc agctgtagcg tgatgcacga
     2041 agctctgcac aaccactaca cccagaaatc tctcagcctg agtcccggca agggcggcgg
     2101 cggcagcggc gggggcggca gcggcggcgg tggatctggc ggaggcggct ccgcccctga
     2161 gcttctgggc ggcccttctg tgttcctgtt ccctccaaag cccaaggaca cactgatgat
     2221 cagcagaacc cctgaagtga cctgtgtggt ggtggacgtg tctcatgaag atcctgaagt
     2281 caaattcaac tggtacgtgg acggggtgga agtgcacaat gccaaaacta agcctcggga
     2341 agagcagtac aatagcacat accgggtggt gtctgtactg accgtgctgc atcaggactg
     2401 gctgaacggc aaggaataca agtgcaaggt gagcaacaag gcactgccag ctcccatcga
     2461 aaagaccatc tccaaggcta agggccagcc tagagagcct caggtgtaca ccctgcctcc
     2521 tagcagagat gaactgacaa aaaaccaggt gagcctgaca tgtctggtta agggctttta
     2581 cccctctgac attgccgtgg aatgggagtc taatggacaa cccgaaaaca actacaagac
     2641 cacaccacct gtgctggata gcgacggcag cttcttcctg accagcaagc tgaccgtgga
     2701 taagtccaga tggcagcagg gcaatgtgtt cagctgcagc gtgatgcacg aggctctgca
     2761 caaccactac acccagaaaa gcctgtccct gagtccaggc aagggcggct ctggcggagg
     2821 aggatctggc ggcctggaag tcctcttcca gggccctctg tgtcctttcg acgaggtgtt
     2881 taacgccaca agattcgcca gcgtttatgc ctggaataga aaaagaatct ctaactgcgt
     2941 ggctgactac agcgtgctgt ataacctggc cccttttttc accttcaagt gctacggcgt
     3001 ttcccctaca aaactgaacg acctgtgctt cacgaacgtg tatgccgaca gcttcgtgat
     3061 ccggggagat gaggtgcggc agatcgcccc tggccaaaca ggcaacatcg ccgattacaa
     3121 ctacaagctg cccgacgact tcacaggatg tgtgatcgcc tggaacagca acaagctcga
     3181 cagcaaagtg agcggaaact acaactacct gtacagactg tttagaaaga gcaatctgaa
     3241 gcccttcgag agagatatca gcaccgagat ctaccaggcc ggcaacaaac cgtgcaacgg
     3301 cgtggccggc ttcaactgct acttccccct gagaagctac tctttccggc ctacctacgg
     3361 cgtgggccac cagccttaca gagtggtggt gctgagcttc gagctgctgc acgcccctgc
     3421 caccgtgtgc ggaggcggcg gcctggtgcc ccagcagagc ggcggactga acgacatctt
     3481 cgaggcccag aagatcgagt ggcacgaggg ctgatgagga tccagatctg ctgtgccttc
     3541 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3601 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3661 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3721 tagcaggcat gctggggatg cggtgggctc tatgggtacc caggtgctga agaattgacc
     3781 cggttcctcc tgggccagaa agaagcaggc acatcccctt ctctgtgaca caccctgtcc
     3841 acgcccctgg ttcttagttc cagccccact cataggacac tcatagctca ggagggctcc
     3901 gccttcaatc ccacccgcta aagtacttgg agcggtctct ccctccctca tcagcccacc
     3961 aaaccaaacc tagcctccaa gagtgggaag aaattaaagc aagataggct attaagtgca
     4021 gagggagaga aaatgcctcc aacatgtgag gaagtaatga gagaaatcat agaattttaa
     4081 ggccatgatt taaggccatc atggccttaa tcttccgctt cctcgctcac tgactcgctg
     4141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5101 cgttcatcca tagttgcctg actcgggggg ggggggcgct gaggtctgcc tcgtgaagaa
     5161 ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag
     5221 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
     5281 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     5341 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt
     5401 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     5461 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     5521 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     5581 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     5641 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     5701 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     5761 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     5821 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     5881 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     5941 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     6001 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     6061 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     6121 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     6181 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     6241 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     6301 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc cccccccccc
     6361 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6421 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6481 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6541 gtc
//
