LOCUS       sequence-347179-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the B.1.1.7 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.1.1.7-AVI
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..87
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(125..147)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     247..266
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        417..796
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        797..1000
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     951..971
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     997..1021
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2930..2953
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3851..3895
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3918..4142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4509..4526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4680..5268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4760..4779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5810..6625)
                     /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(5810..6625)
                     /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     6533..6552
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6617..6636
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
       61 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
      121 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      181 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
      241 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
      301 acagatgcgt aaggagaaaa taccgcatca gattggctat tggccattgc atacgttgta
      361 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca
      421 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      481 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      541 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      601 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      661 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      721 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      781 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      841 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      901 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      961 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     1021 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     1081 cctccatcgg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     1141 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     1201 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     1261 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctagtta
     1321 acggtggagg gcagtgtagt ctgagcagta ctcgttgctg ccgcgcgcgc caccagacat
     1381 aatagctgac agactaacag actgttcctt tccatgggtc ttttctgcag tcaccgtcgt
     1441 cgacacgtgt gatcagatat cgcggccgct ctagaccacc atgggatggt catgtatcat
     1501 cctttttcta gtagcaactg caaccggtgt acattccgca ccagagctgc tgggaggacc
     1561 tagcgtgttc ctgtttcccc ctaagcctaa ggacaccctg atgatcagcc ggaccccaga
     1621 ggtgacatgc gtggtggtgg acgtgtccca cgaggacccc gaggtgaagt tcaactggta
     1681 cgtggatggc gtggaggtgc acaatgccaa gaccaagcca agggaggagc agtacaactc
     1741 tacctatcgc gtggtgagcg tgctgacagt gctgcaccag gactggctga acggcaagga
     1801 gtacaagtgt aaggtgtcta ataaggccct gcctgcccca atcgagaaga ccatcagcaa
     1861 ggcaaaggga cagccaaggg agcctcaggt gtataccctg ccacccagcc gcgacgagct
     1921 gacaaagaac caggtgtccc tgtactgcct ggtgaagggc ttctatcctt ccgatatcgc
     1981 cgtggagtgg gagtctaatg gccagccaga gaacaattac aagaccacac ctccagtgct
     2041 ggactccgat ggctctttct ttctgtattc caagctgacc gtggataaga gccggtggca
     2101 gcagggcaac gtgtttagct gttccgtgat gcacgaggcc ctgcacaatc actacacaca
     2161 gaagtctctg agcctgtccc ccggcaaggg aggaggaggc agcggcggag gaggctccgg
     2221 aggcggcggc tctggcggcg gcggcagcgc ccctgaactg ctgggaggac caagcgtgtt
     2281 cctgtttcca cctaagccca aagataccct gatgatctcc aggacccctg aagtcacctg
     2341 cgtggtcgtg gacgtgtctc atgaggaccc cgaagtcaag ttcaactggt acgtggacgg
     2401 agtcgaagtc cataacgcca agaccaagcc tagagaagag cagtacaact ccacctatag
     2461 agtggtgtct gtgctgacag tgctgcatca ggattggctg aatggcaagg aatataagtg
     2521 taaggtgagc aataaggccc tgcccgcccc tatcgagaag accatctcta aagccaaagg
     2581 acagccaagg gagccacagg tgtatacact gccacccagc cgggacgagc tgaccaagaa
     2641 ccaggtgagc ctgacatgtc tggtcaaggg cttctacccc agtgacattg ctgtggagtg
     2701 ggagtccaat ggccagcctg agaacaatta taaaactacc ccaccagtgc tggacagcga
     2761 tggctccttc tttctgacca gcaagctgac agtggataag tcccgctggc agcagggaaa
     2821 cgtgttttct tgtagcgtga tgcatgaggc cctgcataac cactacaccc agaagtccct
     2881 gtctctgagc ccaggcaagg gaggcagcgg aggcggaggc agcggaggcc tggaggtgct
     2941 gtttcagggc cctcagtgcg tgaacctgac cacccgaacc cagctcccac cagcctacac
     3001 caacagcttt acacggggcg tgtactaccc tgacaaggtg ttcagatcta gcgtcctgca
     3061 cagcactcag gacctcttcc tgccgttctt cagcaacgtg acatggttcc acgccatcag
     3121 cggcacaaac ggaaccaagc ggtttgataa ccccgtcctg ccattcaatg atggagttta
     3181 cttcgccagt accgagaaga gtaacatcat ccggggctgg atcttcggca ccaccctgga
     3241 tagcaaaaca cagagcctcc tgatcgtgaa caatgccacg aacgtcgtga tcaaggtgtg
     3301 cgagttccag ttttgcaatg atcctttcct gggtgtgtac cacaagaaca acaagagctg
     3361 gatggaaagc gagttcagag tctacagcag cgccaacaac tgcacattcg agtacgtctc
     3421 tcagcctttt ctgatggacc ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt
     3481 cgtgttcaag aacatcgacg gctacttcaa aatctactcc aagcacacac ccatcaacct
     3541 ggtccgggac ctccctcagg gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg
     3601 catcaacata acgcggttcc aaaccctgct ggccctgcat agatcctacc tgactcctgg
     3661 cgacagcagc agcggatgga ccgccggagc tgcagcctac tatgtgggct acctgcaacc
     3721 tagaaccttc ctgctgaagt acaacgagaa cggcacaatc acagacgccg tcgactgcgc
     3781 cctggaccct ctctctgaga ccaagtgtac actgaagagc ggaggaggcc tggtgccaca
     3841 gcagagcgga ggcctgaacg acatcttcga ggcccagaag atcgagtggc acgagggctg
     3901 atgaggatcc agatctgctg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3961 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4021 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4081 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4141 gggtacccag gtgctgaaga attgacccgg ttcctcctgg gccagaaaga agcaggcaca
     4201 tccccttctc tgtgacacac cctgtccacg cccctggttc ttagttccag ccccactcat
     4261 aggacactca tagctcagga gggctccgcc ttcaatccca cccgctaaag tacttggagc
     4321 ggtctctccc tccctcatca gcccaccaaa ccaaacctag cctccaagag tgggaagaaa
     4381 ttaaagcaag ataggctatt aagtgcagag ggagagaaaa tgcctccaac atgtgaggaa
     4441 gtaatgagag aaatcataga attttaaggc catgatttaa ggccatcatg gccttaatct
     4501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5101 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact cggggggggg
     5521 gggcgctgag gtctgcctcg tgaagaaggt gttgctgact cataccaggc ctgaatcgcc
     5581 ccatcatcca gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac
     5641 cagttggtga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc
     5701 gtgatctgat ccttcaactc agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc
     5761 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     5821 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     5881 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     5941 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     6001 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     6061 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     6121 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     6181 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     6241 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     6301 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     6361 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     6421 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     6481 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     6541 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag
     6601 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     6661 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     6721 acacaacgtg gctttccccc cccccccatt attgaagcat ttatcagggt tattgtctca
     6781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgc
//
