LOCUS       sequence-347186-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain containing E154K mutation 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-E154K-AVI
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..73
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(111..133)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     233..252
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        403..782
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        783..986
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     937..957
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     983..1007
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2916..2939
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3846..3890
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3913..4137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4504..4521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4675..5263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4755..4774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5805..6620)
                     /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(5805..6620)
                     /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     6528..6547
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6612..6631
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
       61 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
      121 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
      181 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
      241 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
      301 agaaaatacc gcatcagatt ggctattggc cattgcatac gttgtatcca tatcataata
      361 tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta
      421 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      481 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      541 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      601 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      661 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      721 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      781 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      841 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      901 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      961 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     1021 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc catcggctcg
     1081 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     1141 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     1201 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     1261 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tagttaacgg tggagggcag
     1321 tgtagtctga gcagtactcg ttgctgccgc gcgcgccacc agacataata gctgacagac
     1381 taacagactg ttcctttcca tgggtctttt ctgcagtcac cgtcgtcgac acgtgtgatc
     1441 agatatcgcg gccgctctag accaccatgg gatggtcatg tatcatcctt tttctagtag
     1501 caactgcaac cggtgtacat tccgcaccag agctgctggg aggacctagc gtgttcctgt
     1561 ttccccctaa gcctaaggac accctgatga tcagccggac cccagaggtg acatgcgtgg
     1621 tggtggacgt gtcccacgag gaccccgagg tgaagttcaa ctggtacgtg gatggcgtgg
     1681 aggtgcacaa tgccaagacc aagccaaggg aggagcagta caactctacc tatcgcgtgg
     1741 tgagcgtgct gacagtgctg caccaggact ggctgaacgg caaggagtac aagtgtaagg
     1801 tgtctaataa ggccctgcct gccccaatcg agaagaccat cagcaaggca aagggacagc
     1861 caagggagcc tcaggtgtat accctgccac ccagccgcga cgagctgaca aagaaccagg
     1921 tgtccctgta ctgcctggtg aagggcttct atccttccga tatcgccgtg gagtgggagt
     1981 ctaatggcca gccagagaac aattacaaga ccacacctcc agtgctggac tccgatggct
     2041 ctttctttct gtattccaag ctgaccgtgg ataagagccg gtggcagcag ggcaacgtgt
     2101 ttagctgttc cgtgatgcac gaggccctgc acaatcacta cacacagaag tctctgagcc
     2161 tgtcccccgg caagggagga ggaggcagcg gcggaggagg ctccggaggc ggcggctctg
     2221 gcggcggcgg cagcgcccct gaactgctgg gaggaccaag cgtgttcctg tttccaccta
     2281 agcccaaaga taccctgatg atctccagga cccctgaagt cacctgcgtg gtcgtggacg
     2341 tgtctcatga ggaccccgaa gtcaagttca actggtacgt ggacggagtc gaagtccata
     2401 acgccaagac caagcctaga gaagagcagt acaactccac ctatagagtg gtgtctgtgc
     2461 tgacagtgct gcatcaggat tggctgaatg gcaaggaata taagtgtaag gtgagcaata
     2521 aggccctgcc cgcccctatc gagaagacca tctctaaagc caaaggacag ccaagggagc
     2581 cacaggtgta tacactgcca cccagccggg acgagctgac caagaaccag gtgagcctga
     2641 catgtctggt caagggcttc taccccagtg acattgctgt ggagtgggag tccaatggcc
     2701 agcctgagaa caattataaa actaccccac cagtgctgga cagcgatggc tccttctttc
     2761 tgaccagcaa gctgacagtg gataagtccc gctggcagca gggaaacgtg ttttcttgta
     2821 gcgtgatgca tgaggccctg cataaccact acacccagaa gtccctgtct ctgagcccag
     2881 gcaagggagg cagcggaggc ggaggcagcg gaggcctgga ggtgctgttt cagggccctc
     2941 agtgcgtgaa cctgaccacc cgaacccagc tcccaccagc ctacaccaac agctttacac
     3001 ggggcgtgta ctaccctgac aaggtgttca gatctagcgt cctgcacagc actcaggacc
     3061 tcttcctgcc gttcttcagc aacgtgacat ggttccacgc catccacgtg agcggcacaa
     3121 acggaaccaa gcggtttgat aaccccgtcc tgccattcaa tgatggagtt tacttcgcca
     3181 gtaccgagaa gagtaacatc atccggggct ggatcttcgg caccaccctg gatagcaaaa
     3241 cacagagcct cctgatcgtg aacaatgcca cgaacgtcgt gatcaaggtg tgcgagttcc
     3301 agttttgcaa tgatcctttc ctgggtgtgt actaccacaa gaacaacaag agctggatga
     3361 agagcgagtt cagagtctac agcagcgcca acaactgcac attcgagtac gtctctcagc
     3421 cttttctgat ggaccttgag gggaaacaag gcaacttcaa gaacctgaga gaattcgtgt
     3481 tcaagaacat cgacggctac ttcaaaatct actccaagca cacacccatc aacctggtcc
     3541 gggacctccc tcagggcttc agcgccctgg aacccctggt cgacctgccc ataggcatca
     3601 acataacgcg gttccaaacc ctgctggccc tgcatagatc ctacctgact cctggcgaca
     3661 gcagcagcgg atggaccgcc ggagctgcag cctactatgt gggctacctg caacctagaa
     3721 ccttcctgct gaagtacaac gagaacggca caatcacaga cgccgtcgac tgcgccctgg
     3781 accctctctc tgagacaaag tgcaccctga agtccggagg aggcctggtg ccacagcaga
     3841 gcggaggcct gaacgacatc ttcgaggccc agaagatcga gtggcacgag ggctgatgag
     3901 gatccagatc tgctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3961 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4021 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4081 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatgggta
     4141 cccaggtgct gaagaattga cccggttcct cctgggccag aaagaagcag gcacatcccc
     4201 ttctctgtga cacaccctgt ccacgcccct ggttcttagt tccagcccca ctcataggac
     4261 actcatagct caggagggct ccgccttcaa tcccacccgc taaagtactt ggagcggtct
     4321 ctccctccct catcagccca ccaaaccaaa cctagcctcc aagagtggga agaaattaaa
     4381 gcaagatagg ctattaagtg cagagggaga gaaaatgcct ccaacatgtg aggaagtaat
     4441 gagagaaatc atagaatttt aaggccatga tttaaggcca tcatggcctt aatcttccgc
     4501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5101 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5341 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5401 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5461 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactcgggg ggggggggcg
     5521 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc
     5581 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt
     5641 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat
     5701 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc
     5761 agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     5821 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     5881 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     5941 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     6001 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     6061 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     6121 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     6181 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     6241 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     6301 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     6361 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     6421 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     6481 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     6541 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt
     6601 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     6661 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     6721 acgtggcttt cccccccccc ccattattga agcatttatc agggttattg tctcatgagc
     6781 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6841 cga
//
