LOCUS       sequence-347184-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain containing T95I-D253G mutations 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-T95I-D253G-AVI
FEATURES             Location/Qualifiers
     source          1..6843
                     /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             3764..3808
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3831..4055
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4422..4439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4593..5181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4673..4692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5723..6538)
                     /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(5723..6538)
                     /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     6446..6465
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6530..6549
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6816..6834
                     /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 cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc ctaaggacac
     1501 cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt cccacgagga
     1561 ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg ccaagaccaa
     1621 gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga cagtgctgca
     1681 ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg ccctgcctgc
     1741 cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc aggtgtatac
     1801 cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact gcctggtgaa
     1861 gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc cagagaacaa
     1921 ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt attccaagct
     1981 gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg tgatgcacga
     2041 ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca agggaggagg
     2101 aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca gcgcccctga
     2161 actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata ccctgatgat
     2221 ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg accccgaagt
     2281 caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca agcctagaga
     2341 agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc atcaggattg
     2401 gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg cccctatcga
     2461 gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata cactgccacc
     2521 cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca agggcttcta
     2581 ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca attataaaac
     2641 taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc tgacagtgga
     2701 taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg aggccctgca
     2761 taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca gcggaggcgg
     2821 aggcagcgga ggcctggagg tgctgtttca gggccctcag tgcgtgaacc tgaccacccg
     2881 aacccagctc ccaccagcct acaccaacag ctttacacgg ggcgtgtact accctgacaa
     2941 ggtgttcaga tctagcgtcc tgcacagcac tcaggacctc ttcctgccgt tcttcagcaa
     3001 cgtgacatgg ttccacgcca tccacgtgag cggcacaaac ggaaccaagc ggtttgataa
     3061 ccccgtcctg ccattcaatg atggagttta cttcgccagt atcgagaaga gtaacatcat
     3121 ccggggctgg atcttcggca ccaccctgga tagcaaaaca cagagcctcc tgatcgtgaa
     3181 caatgccacg aacgtcgtga tcaaggtgtg cgagttccag ttttgcaatg atcctttcct
     3241 gggtgtgtac taccacaaga acaacaagag ctggatggaa agcgagttca gagtctacag
     3301 cagcgccaac aactgcacat tcgagtacgt ctctcagcct tttctgatgg accttgaggg
     3361 gaaacaaggc aacttcaaga acctgagaga attcgtgttc aagaacatcg acggctactt
     3421 caaaatctac tccaagcaca cacccatcaa cctggtccgg gacctccctc agggcttcag
     3481 cgccctggaa cccctggtcg acctgcccat aggcatcaac ataacgcggt tccaaaccct
     3541 gctggccctg catagatcct acctgactcc tggcggcagc agcagcggat ggaccgccgg
     3601 agctgcagcc tactatgtgg gctacctgca acctagaacc ttcctgctga agtacaacga
     3661 gaacggcaca atcacagacg ccgtcgactg cgccctggac cctctctctg agacaaagtg
     3721 caccctgaag tccggaggag gcctggtgcc acagcagagc ggaggcctga acgacatctt
     3781 cgaggcccag aagatcgagt ggcacgaggg ctgatgagga tccagatctg ctgtgccttc
     3841 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3901 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3961 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4021 tagcaggcat gctggggatg cggtgggctc tatgggtacc caggtgctga agaattgacc
     4081 cggttcctcc tgggccagaa agaagcaggc acatcccctt ctctgtgaca caccctgtcc
     4141 acgcccctgg ttcttagttc cagccccact cataggacac tcatagctca ggagggctcc
     4201 gccttcaatc ccacccgcta aagtacttgg agcggtctct ccctccctca tcagcccacc
     4261 aaaccaaacc tagcctccaa gagtgggaag aaattaaagc aagataggct attaagtgca
     4321 gagggagaga aaatgcctcc aacatgtgag gaagtaatga gagaaatcat agaattttaa
     4381 ggccatgatt taaggccatc atggccttaa tcttccgctt cctcgctcac tgactcgctg
     4441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5401 cgttcatcca tagttgcctg actcgggggg ggggggcgct gaggtctgcc tcgtgaagaa
     5461 ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag
     5521 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
     5581 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     5641 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt
     5701 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     5761 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     5821 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     5881 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     5941 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     6001 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     6061 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     6121 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     6181 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     6241 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     6301 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     6361 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     6421 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     6481 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     6541 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     6601 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc cccccccccc
     6661 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6721 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6781 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6841 gtc
//
