LOCUS       sequence-347246-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the B.1.617.1 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 6843)
  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 6843)
  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.1.617.1-AVI
FEATURES             Location/Qualifiers
     source          1..6843
                     /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             3824..3868
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3891..4115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4482..4499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4653..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4733..4752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5783..6598)
                     /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(5783..6598)
                     /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     6506..6525
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6590..6609
                     /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 tgaccacccg
     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 ctggatgaag agcgagttca gagtctacag
     3361 cagcgccaac aactgcacat tcgagtacgt ctctcagcct tttctgatgg accttgaggg
     3421 gaaacaaggc aacttcaaga acctgagaga attcgtgttc aagaacatcg acggctactt
     3481 caaaatctac tccaagcaca cacccatcaa cctggtccgg gacctccctc agggcttcag
     3541 cgccctggaa cccctggtcg acctgcccat aggcatcaac ataacgcggt tccaaaccct
     3601 gctggccctg catagatcct acctgactcc tggcgacagc agcagcggat ggaccgccgg
     3661 agctgcagcc tactatgtgg gctacctgca acctagaacc ttcctgctga agtacaacga
     3721 gaacggcaca atcacagacg ccgtcgactg cgccctggac cctctctctg agacaaagtg
     3781 caccctgaag tccggaggag gcctggtgcc acagcagagc ggaggcctga acgacatctt
     3841 cgaggcccag aagatcgagt ggcacgaggg ctgatgagga tccagatctg ctgtgccttc
     3901 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3961 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4021 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4081 tagcaggcat gctggggatg cggtgggctc tatgggtacc caggtgctga agaattgacc
     4141 cggttcctcc tgggccagaa agaagcaggc acatcccctt ctctgtgaca caccctgtcc
     4201 acgcccctgg ttcttagttc cagccccact cataggacac tcatagctca ggagggctcc
     4261 gccttcaatc ccacccgcta aagtacttgg agcggtctct ccctccctca tcagcccacc
     4321 aaaccaaacc tagcctccaa gagtgggaag aaattaaagc aagataggct attaagtgca
     4381 gagggagaga aaatgcctcc aacatgtgag gaagtaatga gagaaatcat agaattttaa
     4441 ggccatgatt taaggccatc atggccttaa tcttccgctt cctcgctcac tgactcgctg
     4501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5461 cgttcatcca tagttgcctg actcgggggg ggggggcgct gaggtctgcc tcgtgaagaa
     5521 ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag
     5581 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
     5641 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     5701 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt
     5761 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     5821 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     5881 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     5941 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     6001 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     6061 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     6121 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     6181 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     6241 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     6301 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     6361 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     6421 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     6481 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     6541 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     6601 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     6661 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc cccccccccc
     6721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6841 aag
//
