LOCUS       sequence-347264-        6837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the B.1.618 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 6837)
  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 6837)
  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-NTD-B.1.618-AVI
FEATURES             Location/Qualifiers
     source          1..6837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..63
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(101..123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     223..242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        393..772
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        773..976
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     927..947
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     973..997
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2906..2929
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3830..3874
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3897..4121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4488..4505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4659..5247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4739..4758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5789..6604)
                     /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(5789..6604)
                     /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     6512..6531
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6596..6615
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
       61 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      121 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      181 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      241 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      301 gcatcagatt ggctattggc cattgcatac gttgtatcca tatcataata tgtacattta
      361 tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata
      421 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      481 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      541 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      601 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      661 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      721 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      781 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      841 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      901 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      961 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     1021 ttttgacctc catagaagac accgggaccg atccagcctc catcggctcg catctctcct
     1081 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     1141 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     1201 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     1261 acgctttgcc tgaccctgct tgctcaactc tagttaacgg tggagggcag tgtagtctga
     1321 gcagtactcg ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg
     1381 ttcctttcca tgggtctttt ctgcagtcac cgtcgtcgac acgtgtgatc agatatcgcg
     1441 gccgctctag accaccatgg gatggtcatg tatcatcctt tttctagtag caactgcaac
     1501 cggtgtacat tccgcaccag agctgctggg aggacctagc gtgttcctgt ttccccctaa
     1561 gcctaaggac accctgatga tcagccggac cccagaggtg acatgcgtgg tggtggacgt
     1621 gtcccacgag gaccccgagg tgaagttcaa ctggtacgtg gatggcgtgg aggtgcacaa
     1681 tgccaagacc aagccaaggg aggagcagta caactctacc tatcgcgtgg tgagcgtgct
     1741 gacagtgctg caccaggact ggctgaacgg caaggagtac aagtgtaagg tgtctaataa
     1801 ggccctgcct gccccaatcg agaagaccat cagcaaggca aagggacagc caagggagcc
     1861 tcaggtgtat accctgccac ccagccgcga cgagctgaca aagaaccagg tgtccctgta
     1921 ctgcctggtg aagggcttct atccttccga tatcgccgtg gagtgggagt ctaatggcca
     1981 gccagagaac aattacaaga ccacacctcc agtgctggac tccgatggct ctttctttct
     2041 gtattccaag ctgaccgtgg ataagagccg gtggcagcag ggcaacgtgt ttagctgttc
     2101 cgtgatgcac gaggccctgc acaatcacta cacacagaag tctctgagcc tgtcccccgg
     2161 caagggagga ggaggcagcg gcggaggagg ctccggaggc ggcggctctg gcggcggcgg
     2221 cagcgcccct gaactgctgg gaggaccaag cgtgttcctg tttccaccta agcccaaaga
     2281 taccctgatg atctccagga cccctgaagt cacctgcgtg gtcgtggacg tgtctcatga
     2341 ggaccccgaa gtcaagttca actggtacgt ggacggagtc gaagtccata acgccaagac
     2401 caagcctaga gaagagcagt acaactccac ctatagagtg gtgtctgtgc tgacagtgct
     2461 gcatcaggat tggctgaatg gcaaggaata taagtgtaag gtgagcaata aggccctgcc
     2521 cgcccctatc gagaagacca tctctaaagc caaaggacag ccaagggagc cacaggtgta
     2581 tacactgcca cccagccggg acgagctgac caagaaccag gtgagcctga catgtctggt
     2641 caagggcttc taccccagtg acattgctgt ggagtgggag tccaatggcc agcctgagaa
     2701 caattataaa actaccccac cagtgctgga cagcgatggc tccttctttc tgaccagcaa
     2761 gctgacagtg gataagtccc gctggcagca gggaaacgtg ttttcttgta gcgtgatgca
     2821 tgaggccctg cataaccact acacccagaa gtccctgtct ctgagcccag gcaagggagg
     2881 cagcggaggc ggaggcagcg gaggcctgga ggtgctgttt cagggccctc agtgcgtgaa
     2941 cctgaccacc cgaacccagc tcccaccagc ctacaccaac agctttacac ggggcgtgta
     3001 ctaccctgac aaggtgttca gatctagcgt cctgcacagc actcaggacc tcttcctgcc
     3061 gttcttcagc aacgtgacat ggttccacgc catccacgtg agcggcacaa acggaaccaa
     3121 gcggtttgat aaccccgtcc tgccattcaa tgatggagtt tacttcgcca gtaccgagaa
     3181 gagtaacatc atccggggct ggatcttcgg caccaccctg gatagcaaaa cacagagcct
     3241 cctgatcgtg aacaatgcca cgaacgtcgt gatcaaggtg tgcgagttcc agttttgcaa
     3301 tgatcctttc ctgggtgtgt acaagaacaa caagagctgg atggaaagcg agttcagagt
     3361 ctacagcagc gccaacaact gcacattcga gtacgtctct cagccttttc tgatggacct
     3421 tgaggggaaa caaggcaact tcaagaacct gagagaattc gtgttcaaga acatcgacgg
     3481 ctacttcaaa atctactcca agcacacacc catcaacctg gtccgggacc tccctcaggg
     3541 cttcagcgcc ctggaacccc tggtcgacct gcccataggc atcaacataa cgcggttcca
     3601 aaccctgctg gccctgcata gatcctacct gactcctggc gacagcagca gcggatggac
     3661 cgccggagct gcagcctact atgtgggcta cctgcaacct agaaccttcc tgctgaagta
     3721 caacgagaac ggcacaatca cagacgccgt cgactgcgcc ctggaccctc tctctgagac
     3781 aaagtgcacc ctgaagtccg gaggaggcct ggtgccacag cagagcggag gcctgaacga
     3841 catcttcgag gcccagaaga tcgagtggca cgagggctga tgaggatcca gatctgctgt
     3901 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3961 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4021 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4081 agacaatagc aggcatgctg gggatgcggt gggctctatg ggtacccagg tgctgaagaa
     4141 ttgacccggt tcctcctggg ccagaaagaa gcaggcacat ccccttctct gtgacacacc
     4201 ctgtccacgc ccctggttct tagttccagc cccactcata ggacactcat agctcaggag
     4261 ggctccgcct tcaatcccac ccgctaaagt acttggagcg gtctctccct ccctcatcag
     4321 cccaccaaac caaacctagc ctccaagagt gggaagaaat taaagcaaga taggctatta
     4381 agtgcagagg gagagaaaat gcctccaaca tgtgaggaag taatgagaga aatcatagaa
     4441 ttttaaggcc atgatttaag gccatcatgg ccttaatctt ccgcttcctc gctcactgac
     4501 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4561 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4621 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4681 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4741 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4801 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4861 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4921 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4981 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5041 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5101 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5161 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5221 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5281 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5341 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5401 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5461 ctatttcgtt catccatagt tgcctgactc gggggggggg ggcgctgagg tctgcctcgt
     5521 gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc catcatccag ccagaaagtg
     5581 agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat tttgaacttt
     5641 tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca
     5701 gcaaaagttc gatttattca acaaagccgc cgtcccgtca agtcagcgta atgctctgcc
     5761 agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     5821 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     5881 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     5941 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     6001 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     6061 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     6121 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     6181 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     6241 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     6301 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     6361 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     6421 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     6481 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     6541 cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc
     6601 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     6661 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcccccc
     6721 ccccccatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6781 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
