LOCUS       sequence-347183-        6896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain containing W152C 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 6896)
  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 6896)
  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-W152C-AVI
FEATURES             Location/Qualifiers
     source          1..6896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..28
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(66..88)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     188..207
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        358..737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        738..941
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     892..912
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     938..962
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2871..2894
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2925..2948
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3855..3899
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3921..4145
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4512..4529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4683..5271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4763..4782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5813..6628)
                     /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(5813..6628)
                     /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     6536..6555
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6620..6639
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
       61 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      121 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
      181 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
      241 cacagatgcg taaggagaaa ataccgcatc agattggcta ttggccattg catacgttgt
      301 atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg ccatgttgac
      361 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      421 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      481 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      541 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      601 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      661 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      721 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      781 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      841 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      901 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      961 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
     1021 gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     1081 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     1141 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     1201 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctagtt
     1261 aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg ccaccagaca
     1321 taatagctga cagactaaca gactgttcct ttccatgggt cttttctgca gtcaccgtcg
     1381 tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg tcatgtatca
     1441 tcctttttct agtagcaact gcaaccggtg tacattccgc accagagctg ctgggaggac
     1501 ctagcgtgtt cctgtttccc cctaagccta aggacaccct gatgatcagc cggaccccag
     1561 aggtgacatg cgtggtggtg gacgtgtccc acgaggaccc cgaggtgaag ttcaactggt
     1621 acgtggatgg cgtggaggtg cacaatgcca agaccaagcc aagggaggag cagtacaact
     1681 ctacctatcg cgtggtgagc gtgctgacag tgctgcacca ggactggctg aacggcaagg
     1741 agtacaagtg taaggtgtct aataaggccc tgcctgcccc aatcgagaag accatcagca
     1801 aggcaaaggg acagccaagg gagcctcagg tgtataccct gccacccagc cgcgacgagc
     1861 tgacaaagaa ccaggtgtcc ctgtactgcc tggtgaaggg cttctatcct tccgatatcg
     1921 ccgtggagtg ggagtctaat ggccagccag agaacaatta caagaccaca cctccagtgc
     1981 tggactccga tggctctttc tttctgtatt ccaagctgac cgtggataag agccggtggc
     2041 agcagggcaa cgtgtttagc tgttccgtga tgcacgaggc cctgcacaat cactacacac
     2101 agaagtctct gagcctgtcc cccggcaagg gaggaggagg cagcggcgga ggaggctccg
     2161 gaggcggcgg ctctggcggc ggcggcagcg cccctgaact gctgggagga ccaagcgtgt
     2221 tcctgtttcc acctaagccc aaagataccc tgatgatctc caggacccct gaagtcacct
     2281 gcgtggtcgt ggacgtgtct catgaggacc ccgaagtcaa gttcaactgg tacgtggacg
     2341 gagtcgaagt ccataacgcc aagaccaagc ctagagaaga gcagtacaac tccacctata
     2401 gagtggtgtc tgtgctgaca gtgctgcatc aggattggct gaatggcaag gaatataagt
     2461 gtaaggtgag caataaggcc ctgcccgccc ctatcgagaa gaccatctct aaagccaaag
     2521 gacagccaag ggagccacag gtgtatacac tgccacccag ccgggacgag ctgaccaaga
     2581 accaggtgag cctgacatgt ctggtcaagg gcttctaccc cagtgacatt gctgtggagt
     2641 gggagtccaa tggccagcct gagaacaatt ataaaactac cccaccagtg ctggacagcg
     2701 atggctcctt ctttctgacc agcaagctga cagtggataa gtcccgctgg cagcagggaa
     2761 acgtgttttc ttgtagcgtg atgcatgagg ccctgcataa ccactacacc cagaagtccc
     2821 tgtctctgag cccaggcaag ggaggcagcg gaggcggagg cagcggaggc ctggaggtgc
     2881 tgtttcaagg tccaggtggt tctggcggag gtggatctgg tggtctcgaa gtcctcttcc
     2941 agggccctca gtgcgtgaac ctgaccaccc gaacccagct cccaccagcc tacaccaaca
     3001 gctttacacg gggcgtgtac taccctgaca aggtgttcag atctagcgtc ctgcacagca
     3061 ctcaggacct cttcctgccg ttcttcagca acgtgacatg gttccacgcc atccacgtga
     3121 gcggcacaaa cggaaccaag cggtttgata accccgtcct gccattcaat gatggagttt
     3181 acttcgccag taccgagaag agtaacatca tccggggctg gatcttcggc accaccctgg
     3241 atagcaaaac acagagcctc ctgatcgtga acaatgccac gaacgtcgtg atcaaggtgt
     3301 gcgagttcca gttttgcaat gatcctttcc tgggtgtgta ctaccacaag aacaacaaga
     3361 gctgcatgga aagcgagttc agagtctaca gcagcgccaa caactgcaca ttcgagtacg
     3421 tctctcagcc ttttctgatg gaccttgagg ggaaacaagg caacttcaag aacctgagag
     3481 aattcgtgtt caagaacatc gacggctact tcaaaatcta ctccaagcac acacccatca
     3541 acctggtccg ggacctccct cagggcttca gcgccctgga acccctggtc gacctgccca
     3601 taggcatcaa cataacgcgg ttccaaaccc tgctggccct gcatagatcc tacctgactc
     3661 ctggcgacag cagcagcgga tggaccgccg gagctgcagc ctactatgtg ggctacctgc
     3721 aacctagaac cttcctgctg aagtacaacg agaacggcac aatcacagac gccgtcgact
     3781 gcgccctgga ccctctctct gagaccaagt gtacactgaa gagcggagga ggcctggtgc
     3841 cacagcagag cggaggcctg aacgacatct tcgaggccca gaagatcgag tggcacgagg
     3901 gctgatgagg atcagatctg ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3961 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4021 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4081 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4141 tatgggtacc caggtgctga agaattgacc cggttcctcc tgggccagaa agaagcaggc
     4201 acatcccctt ctctgtgaca caccctgtcc acgcccctgg ttcttagttc cagccccact
     4261 cataggacac tcatagctca ggagggctcc gccttcaatc ccacccgcta aagtacttgg
     4321 agcggtctct ccctccctca tcagcccacc aaaccaaacc tagcctccaa gagtgggaag
     4381 aaattaaagc aagataggct attaagtgca gagggagaga aaatgcctcc aacatgtgag
     4441 gaagtaatga gagaaatcat agaattttaa ggccatgatt taaggccatc atggccttaa
     4501 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4561 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4621 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4681 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4801 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4861 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4921 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4981 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5041 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5101 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5161 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5221 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5281 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5341 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5401 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5461 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actcgggggg
     5521 ggggggcgct gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc
     5581 gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg
     5641 gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga
     5701 tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccgccgtccc
     5761 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     5821 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     5881 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     5941 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     6001 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     6061 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     6121 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     6181 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     6241 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     6301 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     6361 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     6421 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     6481 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     6541 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
     6601 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     6661 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     6721 gagacacaac gtggctttcc cccccccccc attattgaag catttatcag ggttattgtc
     6781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6841 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acatta
//
