LOCUS       sequence-347180-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the B.1.351 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.1.351-AVI
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..38
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(76..98)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     198..217
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        368..747
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        748..951
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     902..922
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     948..972
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2881..2904
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3802..3846
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3869..4093
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4460..4477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4631..5219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4711..4730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5761..6576)
                     /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(5761..6576)
                     /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     6484..6503
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6568..6587
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
       61 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
      121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
      181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
      241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc agattggcta ttggccattg
      301 catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg
      361 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      421 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      481 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      541 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      601 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      661 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      721 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      781 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      841 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      901 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      961 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     1021 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     1081 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     1141 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     1201 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     1261 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg
     1321 ccaccagaca taatagctga cagactaaca gactgttcct ttccatgggt cttttctgca
     1381 gtcaccgtcg tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg
     1441 tcatgtatca tcctttttct agtagcaact gcaaccggtg tacattccgc accagagctg
     1501 ctgggaggac ctagcgtgtt cctgtttccc cctaagccta aggacaccct gatgatcagc
     1561 cggaccccag aggtgacatg cgtggtggtg gacgtgtccc acgaggaccc cgaggtgaag
     1621 ttcaactggt acgtggatgg cgtggaggtg cacaatgcca agaccaagcc aagggaggag
     1681 cagtacaact ctacctatcg cgtggtgagc gtgctgacag tgctgcacca ggactggctg
     1741 aacggcaagg agtacaagtg taaggtgtct aataaggccc tgcctgcccc aatcgagaag
     1801 accatcagca aggcaaaggg acagccaagg gagcctcagg tgtataccct gccacccagc
     1861 cgcgacgagc tgacaaagaa ccaggtgtcc ctgtactgcc tggtgaaggg cttctatcct
     1921 tccgatatcg ccgtggagtg ggagtctaat ggccagccag agaacaatta caagaccaca
     1981 cctccagtgc tggactccga tggctctttc tttctgtatt ccaagctgac cgtggataag
     2041 agccggtggc agcagggcaa cgtgtttagc tgttccgtga tgcacgaggc cctgcacaat
     2101 cactacacac agaagtctct gagcctgtcc cccggcaagg gaggaggagg cagcggcgga
     2161 ggaggctccg gaggcggcgg ctctggcggc ggcggcagcg cccctgaact gctgggagga
     2221 ccaagcgtgt tcctgtttcc acctaagccc aaagataccc tgatgatctc caggacccct
     2281 gaagtcacct gcgtggtcgt ggacgtgtct catgaggacc ccgaagtcaa gttcaactgg
     2341 tacgtggacg gagtcgaagt ccataacgcc aagaccaagc ctagagaaga gcagtacaac
     2401 tccacctata gagtggtgtc tgtgctgaca gtgctgcatc aggattggct gaatggcaag
     2461 gaatataagt gtaaggtgag caataaggcc ctgcccgccc ctatcgagaa gaccatctct
     2521 aaagccaaag gacagccaag ggagccacag gtgtatacac tgccacccag ccgggacgag
     2581 ctgaccaaga accaggtgag cctgacatgt ctggtcaagg gcttctaccc cagtgacatt
     2641 gctgtggagt gggagtccaa tggccagcct gagaacaatt ataaaactac cccaccagtg
     2701 ctggacagcg atggctcctt ctttctgacc agcaagctga cagtggataa gtcccgctgg
     2761 cagcagggaa acgtgttttc ttgtagcgtg atgcatgagg ccctgcataa ccactacacc
     2821 cagaagtccc tgtctctgag cccaggcaag ggaggcagcg gaggcggagg cagcggaggc
     2881 ctggaggtgc tgtttcaggg ccctcagtgc gtgaacttca ccacccgaac ccagctccca
     2941 ccagcctaca ccaacagctt tacacggggc gtgtactacc ctgacaaggt gttcagatct
     3001 agcgtcctgc acagcactca ggacctcttc ctgccgttct tcagcaacgt gacatggttc
     3061 cacgccatcc acgtgagcgg cacaaacgga accaagcggt ttgctaaccc cgtcctgcca
     3121 ttcaatgatg gagtttactt cgccagtacc gagaagagta acatcatccg gggctggatc
     3181 ttcggcacca ccctggatag caaaacacag agcctcctga tcgtgaacaa tgccacgaac
     3241 gtcgtgatca aggtgtgcga gttccagttt tgcaatgatc ctttcctggg tgtgtactac
     3301 cacaagaaca acaagagctg gatggaaagc gagttcagag tctacagcag cgccaacaac
     3361 tgcacattcg agtacgtctc tcagcctttt ctgatggacc ttgaggggaa acaaggcaac
     3421 ttcaagaacc tgagagaatt cgtgttcaag aacatcgacg gctacttcaa aatctactcc
     3481 aagcacacac ccatcaacct ggtccggggc ctccctcagg gcttcagcgc cctggaaccc
     3541 ctggtcgacc tgcccatagg catcaacata acgcggttcc aaaccctgca tatctcctac
     3601 ctgactcctg gcgacagcag cagcggatgg accgccggag ctgcagccta ctatgtgggc
     3661 tacctgcaac ctagaacctt cctgctgaag tacaacgaga acggcacaat cacagacgcc
     3721 gtcgactgcg ccctggaccc tctctctgag acaaagtgca ccctgaagtc cggaggaggc
     3781 ctggtgccac agcagagcgg aggcctgaac gacatcttcg aggcccagaa gatcgagtgg
     3841 cacgagggct gatgaggatc cagatctgct gtgccttcta gttgccagcc atctgttgtt
     3901 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3961 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4021 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4081 gtgggctcta tgggtaccca ggtgctgaag aattgacccg gttcctcctg ggccagaaag
     4141 aagcaggcac atccccttct ctgtgacaca ccctgtccac gcccctggtt cttagttcca
     4201 gccccactca taggacactc atagctcagg agggctccgc cttcaatccc acccgctaaa
     4261 gtacttggag cggtctctcc ctccctcatc agcccaccaa accaaaccta gcctccaaga
     4321 gtgggaagaa attaaagcaa gataggctat taagtgcaga gggagagaaa atgcctccaa
     4381 catgtgagga agtaatgaga gaaatcatag aattttaagg ccatgattta aggccatcat
     4441 ggccttaatc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4501 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4621 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5041 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5281 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5341 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5401 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5461 tcgggggggg ggggcgctga ggtctgcctc gtgaagaagg tgttgctgac tcataccagg
     5521 cctgaatcgc cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt
     5581 tgtaggtgga ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt
     5641 cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc
     5701 gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga
     5761 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     5821 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     5881 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     5941 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     6001 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     6061 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     6121 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     6181 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     6241 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     6301 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     6361 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     6421 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     6481 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     6541 cctcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6601 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     6661 gagattttga gacacaacgt ggctttcccc ccccccccat tattgaagca tttatcaggg
     6721 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6781 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttat
//
