LOCUS       sequence-347181-        6842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain from the P.1 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 6842)
  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 6842)
  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-P.1-AVI
FEATURES             Location/Qualifiers
     source          1..6842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(41..63)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     163..182
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        333..712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        713..916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     867..887
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     913..937
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2846..2869
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3776..3820
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3842..4066
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4433..4450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4604..5192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4684..4703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5734..6549)
                     /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(5734..6549)
                     /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     6457..6476
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6541..6560
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     join(6827..6842,1..3)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
       61 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      121 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      181 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      241 gcatcagatt ggctattggc cattgcatac gttgtatcca tatcataata tgtacattta
      301 tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata
      361 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      421 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      481 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      541 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      601 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      661 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      721 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      781 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      841 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      901 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
      961 ttttgacctc catagaagac accgggaccg atccagcctc catcggctcg catctctcct
     1021 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     1081 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     1141 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     1201 acgctttgcc tgaccctgct tgctcaactc tagttaacgg tggagggcag tgtagtctga
     1261 gcagtactcg ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg
     1321 ttcctttcca tgggtctttt ctgcagtcac cgtcgtcgac acgtgtgatc agatatcgcg
     1381 gccgctctag accaccatgg gatggtcatg tatcatcctt tttctagtag caactgcaac
     1441 cggtgtacat tccgcaccag agctgctggg aggacctagc gtgttcctgt ttccccctaa
     1501 gcctaaggac accctgatga tcagccggac cccagaggtg acatgcgtgg tggtggacgt
     1561 gtcccacgag gaccccgagg tgaagttcaa ctggtacgtg gatggcgtgg aggtgcacaa
     1621 tgccaagacc aagccaaggg aggagcagta caactctacc tatcgcgtgg tgagcgtgct
     1681 gacagtgctg caccaggact ggctgaacgg caaggagtac aagtgtaagg tgtctaataa
     1741 ggccctgcct gccccaatcg agaagaccat cagcaaggca aagggacagc caagggagcc
     1801 tcaggtgtat accctgccac ccagccgcga cgagctgaca aagaaccagg tgtccctgta
     1861 ctgcctggtg aagggcttct atccttccga tatcgccgtg gagtgggagt ctaatggcca
     1921 gccagagaac aattacaaga ccacacctcc agtgctggac tccgatggct ctttctttct
     1981 gtattccaag ctgaccgtgg ataagagccg gtggcagcag ggcaacgtgt ttagctgttc
     2041 cgtgatgcac gaggccctgc acaatcacta cacacagaag tctctgagcc tgtcccccgg
     2101 caagggagga ggaggcagcg gcggaggagg ctccggaggc ggcggctctg gcggcggcgg
     2161 cagcgcccct gaactgctgg gaggaccaag cgtgttcctg tttccaccta agcccaaaga
     2221 taccctgatg atctccagga cccctgaagt cacctgcgtg gtcgtggacg tgtctcatga
     2281 ggaccccgaa gtcaagttca actggtacgt ggacggagtc gaagtccata acgccaagac
     2341 caagcctaga gaagagcagt acaactccac ctatagagtg gtgtctgtgc tgacagtgct
     2401 gcatcaggat tggctgaatg gcaaggaata taagtgtaag gtgagcaata aggccctgcc
     2461 cgcccctatc gagaagacca tctctaaagc caaaggacag ccaagggagc cacaggtgta
     2521 tacactgcca cccagccggg acgagctgac caagaaccag gtgagcctga catgtctggt
     2581 caagggcttc taccccagtg acattgctgt ggagtgggag tccaatggcc agcctgagaa
     2641 caattataaa actaccccac cagtgctgga cagcgatggc tccttctttc tgaccagcaa
     2701 gctgacagtg gataagtccc gctggcagca gggaaacgtg ttttcttgta gcgtgatgca
     2761 tgaggccctg cataaccact acacccagaa gtccctgtct ctgagcccag gcaagggagg
     2821 cagcggaggc ggaggcagcg gaggcctgga ggtgctgttt cagggccccc agtgcgtgaa
     2881 cttcaccaac cgaacccagc tcccaagcgc ctacaccaac agctttacac ggggcgtgta
     2941 ctaccctgac aaggtgttca gatctagcgt cctgcacagc actcaggacc tcttcctgcc
     3001 gttcttcagc aacgtgacat ggttccacgc catccacgtg agcggcacaa acggaaccaa
     3061 gcggtttgat aaccccgtcc tgccattcaa tgatggagtt tacttcgcca gtaccgagaa
     3121 gagtaacatc atccggggct ggatcttcgg caccaccctg gatagcaaaa cacagagcct
     3181 cctgatcgtg aacaatgcca cgaacgtcgt gatcaaggtg tgcgagttcc agttttgcaa
     3241 ttatcctttc ctgggtgtgt actaccacaa gaacaacaag agctggatgg aaagcgagtt
     3301 cagagtctac agcagcgcca acaactgcac attcgagtac gtctctcagc cttttctgat
     3361 ggaccttgag gggaaacaag gcaacttcaa gaacctgagc gaattcgtgt tcaagaacat
     3421 cgacggctac ttcaaaatct actccaagca cacacccatc aacctggtcc gggacctccc
     3481 tcagggcttc agcgccctgg aacccctggt cgacctgccc ataggcatca acataacgcg
     3541 gttccaaacc ctgctggccc tgcatagatc ctacctgact cctggcgaca gcagcagcgg
     3601 atggaccgcc ggagctgcag cctactatgt gggctacctg caacctagaa ccttcctgct
     3661 gaagtacaac gagaacggca caatcacaga cgccgtcgac tgcgccctgg accctctctc
     3721 tgagaccaag tgtacactga agagcggagg aggcctggtg ccacagcaga gcggaggcct
     3781 gaacgacatc ttcgaggccc agaagatcga gtggcacgag ggctgatgag gatcagatct
     3841 gctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3901 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3961 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4021 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatgggtac ccaggtgctg
     4081 aagaattgac ccggttcctc ctgggccaga aagaagcagg cacatcccct tctctgtgac
     4141 acaccctgtc cacgcccctg gttcttagtt ccagccccac tcataggaca ctcatagctc
     4201 aggagggctc cgccttcaat cccacccgct aaagtacttg gagcggtctc tccctccctc
     4261 atcagcccac caaaccaaac ctagcctcca agagtgggaa gaaattaaag caagataggc
     4321 tattaagtgc agagggagag aaaatgcctc caacatgtga ggaagtaatg agagaaatca
     4381 tagaatttta aggccatgat ttaaggccat catggcctta atcttccgct tcctcgctca
     4441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5401 tctgtctatt tcgttcatcc atagttgcct gactcggggg gggggggcgc tgaggtctgc
     5461 ctcgtgaaga aggtgttgct gactcatacc aggcctgaat cgccccatca tccagccaga
     5521 aagtgaggga gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga
     5581 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
     5641 actcagcaaa agttcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct
     5701 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     5761 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     5821 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     5881 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     5941 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     6001 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     6061 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     6121 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     6181 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     6241 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     6301 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     6361 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     6421 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     6481 taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat
     6541 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     6601 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttc
     6661 cccccccccc cattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6781 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6841 ga
//
