LOCUS       sequence-361463-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the BA.2 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-BA.2-AVI
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(54..869)
                     /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(54..869)
                     /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     777..796
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     861..880
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1147..1165
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1203..1225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1325..1344
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1495..1874
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1875..2078
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2029..2049
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2075..2099
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4008..4031
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4929..4973
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    4996..5220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5587..5604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5758..6346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5838..5857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
       61 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
      121 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
      181 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
      241 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
      301 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
      361 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
      421 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
      481 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
      541 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
      601 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
      661 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
      721 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
      781 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
      841 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
      901 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
      961 tgagacacaa cgtggctttn cccccccccc cattattgaa gcatttatca gggttattgt
     1021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1081 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     1141 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     1201 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     1261 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     1321 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     1381 agatgcgtaa ggagaaaata ccgcatcaga ttggctattg gccattgcat acgttgtatc
     1441 catatcataa tatgtacatt tatattggct catgtccaac attaccgcca tgttgacatt
     1501 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     1561 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     1621 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     1681 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1741 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1801 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1861 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     1921 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     1981 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2041 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     2101 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     2161 tccatcggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     2221 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     2281 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     2341 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctagttaac
     2401 ggtggagggc agtgtagtct gagcagtact cgttgctgcc gcgcgcgcca ccagacataa
     2461 tagctgacag actaacagac tgttcctttc catgggtctt ttctgcagtc accgtcgtcg
     2521 acacgtgtga tcagatatcg cggccgctct agaccaccat gggatggtca tgtatcatcc
     2581 tttttctagt agcaactgca accggtgtac attccgcacc agagctgctg ggaggaccta
     2641 gcgtgttcct gtttccccct aagcctaagg acaccctgat gatcagccgg accccagagg
     2701 tgacatgcgt ggtggtggac gtgtcccacg aggaccccga ggtgaagttc aactggtacg
     2761 tggatggcgt ggaggtgcac aatgccaaga ccaagccaag ggaggagcag tacaactcta
     2821 cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga ctggctgaac ggcaaggagt
     2881 acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat cgagaagacc atcagcaagg
     2941 caaagggaca gccaagggag cctcaggtgt ataccctgcc acccagccgc gacgagctga
     3001 caaagaacca ggtgtccctg tactgcctgg tgaagggctt ctatccttcc gatatcgccg
     3061 tggagtggga gtctaatggc cagccagaga acaattacaa gaccacacct ccagtgctgg
     3121 actccgatgg ctctttcttt ctgtattcca agctgaccgt ggataagagc cggtggcagc
     3181 agggcaacgt gtttagctgt tccgtgatgc acgaggccct gcacaatcac tacacacaga
     3241 agtctctgag cctgtccccc ggcaagggag gaggaggcag cggcggagga ggctccggag
     3301 gcggcggctc tggcggcggc ggcagcgccc ctgaactgct gggaggacca agcgtgttcc
     3361 tgtttccacc taagcccaaa gataccctga tgatctccag gacccctgaa gtcacctgcg
     3421 tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt caactggtac gtggacggag
     3481 tcgaagtcca taacgccaag accaagccta gagaagagca gtacaactcc acctatagag
     3541 tggtgtctgt gctgacagtg ctgcatcagg attggctgaa tggcaaggaa tataagtgta
     3601 aggtgagcaa taaggccctg cccgccccta tcgagaagac catctctaaa gccaaaggac
     3661 agccaaggga gccacaggtg tatacactgc cacccagccg ggacgagctg accaagaacc
     3721 aggtgagcct gacatgtctg gtcaagggct tctaccccag tgacattgct gtggagtggg
     3781 agtccaatgg ccagcctgag aacaattata aaactacccc accagtgctg gacagcgatg
     3841 gctccttctt tctgaccagc aagctgacag tggataagtc ccgctggcag cagggaaacg
     3901 tgttttcttg tagcgtgatg catgaggccc tgcataacca ctacacccag aagtccctgt
     3961 ctctgagccc aggcaaggga ggcagcggag gcggaggcag cggaggcctg gaggtgctgt
     4021 ttcagggccc tcagtgcgtg aacctgatca cccgaaccca gagctacacc aacagcttta
     4081 cacggggcgt gtactaccct gacaaggtgt tcagatctag cgtcctgcac agcactcagg
     4141 acctcttcct gccgttcttc agcaacgtga catggttcca cgccatccac gtgagcggca
     4201 caaacggaac caagcggttt gataaccccg tcctgccatt caatgatgga gtttacttcg
     4261 ccagtaccga gaagagtaac atcatccggg gctggatctt cggcaccacc ctggatagca
     4321 aaacacagag cctcctgatc gtgaacaatg ccacgaacgt cgtgatcaag gtgtgcgagt
     4381 tccagttttg caatgatcct ttcctggatg tgtactacca caagaacaac aagagctgga
     4441 tggaaagcga gttcagagtc tacagcagcg ccaacaactg cacattcgag tacgtctctc
     4501 agccttttct gatggacctt gaggggaaac aaggcaactt caagaacctg agagaattcg
     4561 tgttcaagaa catcgacggc tacttcaaaa tctactccaa gcacacaccc atcaacctgg
     4621 gccgggacct ccctcagggc ttcagcgccc tggaacccct ggtcgacctg cccataggca
     4681 tcaacataac gcggttccaa accctgctgg ccctgcatag atcctacctg actcctggcg
     4741 acagcagcag cggatggacc gccggagctg cagcctacta tgtgggctac ctgcaaccta
     4801 gaaccttcct gctgaagtac aacgagaacg gcacaatcac agacgccgtc gactgcgccc
     4861 tggaccctct ctctgagaca aagtgcaccc tgaagtccgg aggaggcctg gtgccacagc
     4921 agagcggagg cctgaacgac atcttcgagg cccagaagat cgagtggcac gagggctgat
     4981 gaggatccag atctgctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5041 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5101 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5161 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5221 gtacccaggt gctgaagaat tgacccggtt cctcctgggc cagaaagaag caggcacatc
     5281 cccttctctg tgacacaccc tgtccacgcc cctggttctt agttccagcc ccactcatag
     5341 gacactcata gctcaggagg gctccgcctt caatcccacc cgctaaagta cttggagcgg
     5401 tctctccctc cctcatcagc ccaccaaacc aaacctagcc tccaagagtg ggaagaaatt
     5461 aaagcaagat aggctattaa gtgcagaggg agagaaaatg cctccaacat gtgaggaagt
     5521 aatgagagaa atcatagaat tttaaggcca tgatttaagg ccatcatggc cttaatcttc
     5581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5941 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6181 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6481 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6541 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcg gggggggggg
     6601 gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca taccaggcct gaatcgcccc
     6661 atcatccagc cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca
     6721 gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt
     6781 gatctgatcc ttcaactcag caaaagttcg atttattcaa caaagccgcc gtcc
//
