LOCUS       sequence-347245-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NTD domain containing G142D-E154K mutations 
            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 6843)
  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 6843)
  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-G142D-E154K-AVI
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(90..905)
                     /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(90..905)
                     /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     813..832
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     897..916
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1183..1201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1239..1261)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1361..1380
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1531..1910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1911..2114
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2065..2085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2111..2135
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4044..4067
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4974..5018
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5041..5265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5632..5649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5803..6391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5883..5902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc
       61 cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
      121 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
      181 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
      241 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
      301 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
      361 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
      421 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
      481 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
      541 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
      601 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
      661 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
      721 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
      781 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
      841 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg
      901 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
      961 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttccccc
     1021 cccccccatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1141 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     1201 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1261 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1321 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1381 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     1441 atcagattgg ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata
     1501 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
     1561 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     1621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     1681 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     1741 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     1801 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1861 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     1921 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     1981 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2041 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2101 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     2161 ttgacctcca tagaagacac cgggaccgat ccagcctcca tcggctcgca tctctccttc
     2221 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     2281 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     2341 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     2401 gctttgcctg accctgcttg ctcaactcta gttaacggtg gagggcagtg tagtctgagc
     2461 agtactcgtt gctgccgcgc gcgccaccag acataatagc tgacagacta acagactgtt
     2521 cctttccatg ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag atatcgcggc
     2581 cgctctagac caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg
     2641 gtgtacattc cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc
     2701 ctaaggacac cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt
     2761 cccacgagga ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg
     2821 ccaagaccaa gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga
     2881 cagtgctgca ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg
     2941 ccctgcctgc cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc
     3001 aggtgtatac cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact
     3061 gcctggtgaa gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc
     3121 cagagaacaa ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt
     3181 attccaagct gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg
     3241 tgatgcacga ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca
     3301 agggaggagg aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca
     3361 gcgcccctga actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata
     3421 ccctgatgat ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg
     3481 accccgaagt caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca
     3541 agcctagaga agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc
     3601 atcaggattg gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg
     3661 cccctatcga gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata
     3721 cactgccacc cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca
     3781 agggcttcta ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca
     3841 attataaaac taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc
     3901 tgacagtgga taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg
     3961 aggccctgca taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca
     4021 gcggaggcgg aggcagcgga ggcctggagg tgctgtttca gggccctcag tgcgtgaacc
     4081 tgaccacccg aacccagctc ccaccagcct acaccaacag ctttacacgg ggcgtgtact
     4141 accctgacaa ggtgttcaga tctagcgtcc tgcacagcac tcaggacctc ttcctgccgt
     4201 tcttcagcaa cgtgacatgg ttccacgcca tccacgtgag cggcacaaac ggaaccaagc
     4261 ggtttgataa ccccgtcctg ccattcaatg atggagttta cttcgccagt accgagaaga
     4321 gtaacatcat ccggggctgg atcttcggca ccaccctgga tagcaaaaca cagagcctcc
     4381 tgatcgtgaa caatgccacg aacgtcgtga tcaaggtgtg cgagttccag ttttgcaatg
     4441 atcctttcct ggacgtgtac taccacaaga acaacaagag ctggatgaag agcgagttca
     4501 gagtctacag cagcgccaac aactgcacat tcgagtacgt ctctcagcct tttctgatgg
     4561 accttgaggg gaaacaaggc aacttcaaga acctgagaga attcgtgttc aagaacatcg
     4621 acggctactt caaaatctac tccaagcaca cacccatcaa cctggtccgg gacctccctc
     4681 agggcttcag cgccctggaa cccctggtcg acctgcccat aggcatcaac ataacgcggt
     4741 tccaaaccct gctggccctg catagatcct acctgactcc tggcgacagc agcagcggat
     4801 ggaccgccgg agctgcagcc tactatgtgg gctacctgca acctagaacc ttcctgctga
     4861 agtacaacga gaacggcaca atcacagacg ccgtcgactg cgccctggac cctctctctg
     4921 agacaaagtg caccctgaag tccggaggag gcctggtgcc acagcagagc ggaggcctga
     4981 acgacatctt cgaggcccag aagatcgagt ggcacgaggg ctgatgagga tccagatctg
     5041 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5101 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5161 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5221 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgggtacc caggtgctga
     5281 agaattgacc cggttcctcc tgggccagaa agaagcaggc acatcccctt ctctgtgaca
     5341 caccctgtcc acgcccctgg ttcttagttc cagccccact cataggacac tcatagctca
     5401 ggagggctcc gccttcaatc ccacccgcta aagtacttgg agcggtctct ccctccctca
     5461 tcagcccacc aaaccaaacc tagcctccaa gagtgggaag aaattaaagc aagataggct
     5521 attaagtgca gagggagaga aaatgcctcc aacatgtgag gaagtaatga gagaaatcat
     5581 agaattttaa ggccatgatt taaggccatc atggccttaa tcttccgctt cctcgctcac
     5641 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5701 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5761 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5821 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5881 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5941 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6001 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6061 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6421 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6601 ctgtctattt cgttcatcca tagttgcctg actcgggggg ggggggcgct gaggtctgcc
     6661 tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa
     6721 agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa
     6781 cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa
     6841 ctc
//
