LOCUS       sequence-347260-        6837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the AY.1 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-AY.1-AVI
FEATURES             Location/Qualifiers
     source          1..6837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..51
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(89..111)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     211..230
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        381..760
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        761..964
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     915..935
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     961..985
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2894..2917
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3818..3862
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3885..4109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4476..4493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4647..5235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4727..4746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5777..6592)
                     /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(5777..6592)
                     /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     6500..6519
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6584..6603
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
       61 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
      121 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      181 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      241 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcagattgg
      301 ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
      361 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      421 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      481 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      541 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      601 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      661 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      721 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      781 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      841 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      901 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      961 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     1021 tagaagacac cgggaccgat ccagcctcca tcggctcgca tctctccttc acgcgcccgc
     1081 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     1141 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     1201 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     1261 accctgcttg ctcaactcta gttaacggtg gagggcagtg tagtctgagc agtactcgtt
     1321 gctgccgcgc gcgccaccag acataatagc tgacagacta acagactgtt cctttccatg
     1381 ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag atatcgcggc cgctctagac
     1441 caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg gtgtacattc
     1501 cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc ctaaggacac
     1561 cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt cccacgagga
     1621 ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg ccaagaccaa
     1681 gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga cagtgctgca
     1741 ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg ccctgcctgc
     1801 cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc aggtgtatac
     1861 cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact gcctggtgaa
     1921 gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc cagagaacaa
     1981 ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt attccaagct
     2041 gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg tgatgcacga
     2101 ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca agggaggagg
     2161 aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca gcgcccctga
     2221 actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata ccctgatgat
     2281 ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg accccgaagt
     2341 caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca agcctagaga
     2401 agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc atcaggattg
     2461 gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg cccctatcga
     2521 gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata cactgccacc
     2581 cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca agggcttcta
     2641 ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca attataaaac
     2701 taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc tgacagtgga
     2761 taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg aggccctgca
     2821 taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca gcggaggcgg
     2881 aggcagcgga ggcctggagg tgctgtttca gggccctcag tgcgtgaacc tgagaacccg
     2941 aacccagctc ccaccagcct acaccaacag ctttacacgg ggcgtgtact accctgacaa
     3001 ggtgttcaga tctagcgtcc tgcacagcac tcaggacctc ttcctgccgt tcttcagcaa
     3061 cgtgacatgg ttccacgcca tccacgtgag cggcacaaac ggaaccaagc ggtttgataa
     3121 ccccgtcctg ccattcaatg atggagttta cttcgccagt atcgagaaga gtaacatcat
     3181 ccggggctgg atcttcggca ccaccctgga tagcaaaaca cagagcctcc tgatcgtgaa
     3241 caatgccacg aacgtcgtga tcaaggtgtg cgagttccag ttttgcaatg atcctttcct
     3301 ggacgtgtac taccacaaga acaacaagag ctggatggaa agcggcgtct acagcagcgc
     3361 caacaactgc acattcgagt acgtctctca gccttttctg atggaccttg aggggaaaca
     3421 aggcaacttc aagaacctga gagaattcgt gttcaagaac atcgacggct acttcaaaat
     3481 ctactccaag cacacaccca tcaacctggt ccgggacctc cctcagggct tcagcgccct
     3541 ggaacccctg gtcgacctgc ccataggcat caacataacg cggttccaaa ccctgctggc
     3601 cctgcataga tcctacctga ctcctggcga cagcagcagc ggactgaccg ccggagctgc
     3661 agcctactat gtgggctacc tgcaacctag aaccttcctg ctgaagtaca acgagaacgg
     3721 cacaatcaca gacgccgtcg actgcgccct ggaccctctc tctgagacaa agtgcaccct
     3781 gaagtccgga ggaggcctgg tgccacagca gagcggaggc ctgaacgaca tcttcgaggc
     3841 ccagaagatc gagtggcacg agggctgatg aggatccaga tctgctgtgc cttctagttg
     3901 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3961 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4021 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4081 gcatgctggg gatgcggtgg gctctatggg tacccaggtg ctgaagaatt gacccggttc
     4141 ctcctgggcc agaaagaagc aggcacatcc ccttctctgt gacacaccct gtccacgccc
     4201 ctggttctta gttccagccc cactcatagg acactcatag ctcaggaggg ctccgccttc
     4261 aatcccaccc gctaaagtac ttggagcggt ctctccctcc ctcatcagcc caccaaacca
     4321 aacctagcct ccaagagtgg gaagaaatta aagcaagata ggctattaag tgcagaggga
     4381 gagaaaatgc ctccaacatg tgaggaagta atgagagaaa tcatagaatt ttaaggccat
     4441 gatttaaggc catcatggcc ttaatcttcc gcttcctcgc tcactgactc gctgcgctcg
     4501 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4561 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5461 tccatagttg cctgactcgg gggggggggg cgctgaggtc tgcctcgtga agaaggtgtt
     5521 gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg
     5581 ttgatgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg
     5641 gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc aaaagttcga
     5701 tttattcaac aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc
     5761 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     5821 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     5881 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     5941 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     6001 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga
     6061 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     6121 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     6181 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     6241 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     6301 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     6361 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     6421 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     6481 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     6541 tggaatttaa tcgcggcctc gagcaagacg tttcccgttg aatatggctc ataacacccc
     6601 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     6661 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttcccccccc ccccattatt
     6721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaag
//
