LOCUS       sequence-354496-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the B.1.1.529 (Omicron) 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 6834)
  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 6834)
  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.1.1.529-AVI
FEATURES             Location/Qualifiers
     source          1..6834
                     /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             3755..3799
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3822..4046
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4413..4430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4584..5172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4664..4683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5714..6529)
                     /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(5714..6529)
                     /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     6437..6456
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6521..6540
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6807..6825
                     /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 cgcccctgag ctgctgggcg gacctagcgt gttcctgttc ccccctaagc ctaaagacac
     1501 cctgatgatc agcagaaccc ctgaagtgac ctgcgtggtc gtggacgtgt ctcacgagga
     1561 ccccgaggtc aagttcaact ggtacgtgga cggcgtggaa gtgcacaacg ctaagaccaa
     1621 gcctcgcgag gaacagtaca atagcacata ccgggtggtg agcgtgctaa ccgtgctcca
     1681 tcaggactgg ctgaacggca aggaatacaa gtgtaaagtg tccaacaagg ccctgcctgc
     1741 accaatcgag aagactatct ctaaggccaa gggccaacct agagagcctc aggtgtacac
     1801 cctgccaccc agcagagacg agctgaccaa gaaccaggtg tctctgtact gcctggtcaa
     1861 gggcttctac ccttctgata tcgccgtgga atgggaaagc aatggccagc cagagaacaa
     1921 ctacaagacc acccctccag tgctcgactc cgacggctct ttcttcctgt acagcaagct
     1981 gaccgtggac aagagcagat ggcagcaggg caacgtgttt tcttgcagcg tgatgcacga
     2041 agctctccac aaccactaca cccagaagag cctgagcctg agccctggca agggtggcgg
     2101 tggctcaggc ggcggcggca gcggcggcgg cggtagcgga ggcggcggca gcgcccctga
     2161 gctcctcggc ggcccctctg tcttcctgtt tccccccaag cctaaggaca ccctgatgat
     2221 ctccagaacc cctgaggtga catgtgtggt ggtcgacgtg agccacgagg accctgaggt
     2281 gaaattcaat tggtatgttg acggcgtgga agtccacaac gccaagacaa agcctagaga
     2341 agagcagtac aacagcacct acagagtggt cagcgttctg accgtgctgc accaggattg
     2401 gctgaacggc aaggagtata agtgcaaggt gagcaacaaa gccctgccag cccccatcga
     2461 aaagacaatt tccaaagcca agggacagcc tagagaacct caggtgtaca ccctgcctcc
     2521 tagccgggac gagctgacca agaaccaggt tagcctgaca tgcctggtga agggctttta
     2581 ccccagcgac atcgccgtgg aatgggagag caacggccag cctgaaaaca attacaaaac
     2641 aacaccacca gtgctggaca gcgatggatc ttttttcctg acatctaagc tgaccgtgga
     2701 taagagccgg tggcagcaag gcaacgtgtt ttcctgtagc gtgatgcacg aggccctcca
     2761 caatcattac acccagaaat ctctgtctct gagccccgga aagggcggca gcggtggcgg
     2821 aggcagcggc ggcctggaag tgctgttcca aggcccccag tgcgtgaacc tgaccacaag
     2881 aacacagctg ccccccgctt atacaaacag cttcaccaga ggcgtctact accctgacaa
     2941 agtgtttaga agcagcgtcc tgcacagcac acaggacctg ttcctgccct tcttcagtaa
     3001 cgtgacctgg ttccacgtga tcagcgggac caacggaaca aaaagattcg acaaccccgt
     3061 gctgcctttt aacgacggcg tgtacttcgc cagcatcgaa aagagcaaca tcatccgggg
     3121 atggatcttc ggcactaccc tggattccaa gacccagagc ctgctgatcg tgaacaacgc
     3181 caccaatgtg gtgatcaagg tgtgcgagtt tcagttctgc aacgaccctt tcctggatca
     3241 caagaacaac aagagctgga tggaatccga gttcagggtg tatagcagcg ccaacaactg
     3301 caccttcgag tacgtgagcc agccttttct gatggacctg gaaggcaaac agggaaattt
     3361 caagaacctg agagagttcg tgttcaagaa catcgacgga tacttcaaga tctacagtaa
     3421 gcacacccct atcatcgtgc gggagcctga ggacctgcct cagggcttct cagccctgga
     3481 acctctggta gatctcccca taggaatcaa catcacccgg ttccagacac tgctggccct
     3541 gcacagaagc tacctgaccc ctggagatag cagcagcggc tggaccgccg gcgccgctgc
     3601 ttactacgtg gggtacctgc agccgcggac cttcctgctg aaatacaatg agaatggcac
     3661 catcactgat gccgtggact gcgccctgga ccctctgagc gagacaaagt gtaccctgaa
     3721 gtctggcggc ggcctggtgc cacaacaaag cggcggcctg aacgacatct tcgaggccca
     3781 gaaaattgag tggcacgagg gctgatgagg atccagatct gctgtgcctt ctagttgcca
     3841 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3901 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3961 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4021 tgctggggat gcggtgggct ctatgggtac ccaggtgctg aagaattgac ccggttcctc
     4081 ctgggccaga aagaagcagg cacatcccct tctctgtgac acaccctgtc cacgcccctg
     4141 gttcttagtt ccagccccac tcataggaca ctcatagctc aggagggctc cgccttcaat
     4201 cccacccgct aaagtacttg gagcggtctc tccctccctc atcagcccac caaaccaaac
     4261 ctagcctcca agagtgggaa gaaattaaag caagataggc tattaagtgc agagggagag
     4321 aaaatgcctc caacatgtga ggaagtaatg agagaaatca tagaatttta aggccatgat
     4381 ttaaggccat catggcctta atcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4441 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4501 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4561 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4621 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4681 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4741 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4801 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4861 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4921 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4981 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5041 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5101 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5161 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5221 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5281 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5341 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5401 atagttgcct gactcggggg gggggggcgc tgaggtctgc ctcgtgaaga aggtgttgct
     5461 gactcatacc aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg
     5521 atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa
     5581 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt
     5641 attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat
     5701 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     5761 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     5821 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     5881 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     5941 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     6001 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     6061 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     6121 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     6181 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     6241 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     6301 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     6361 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     6421 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     6481 aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg
     6541 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     6601 caatgtaaca tcagagattt tgagacacaa cgtggctttc cccccccccc cattattgaa
     6661 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6721 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6781 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
