LOCUS       sequence-347367-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing S477N mutation 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 6558)
  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 6558)
  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-RBD-S477N-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     87..105
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(143..165)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     265..284
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        435..814
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        815..1018
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     969..989
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1015..1039
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2948..2971
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3593..3637
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3660..3884
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4251..4268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4422..5010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4502..4521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5552..6367)
                     /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(5552..6367)
                     /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     6275..6294
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6359..6378
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
       61 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
      121 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
      181 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      241 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      301 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcaga ttggctattg
      361 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
      421 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      481 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      541 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      601 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      661 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      721 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      781 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      841 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      901 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      961 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     1021 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     1081 acaccgggac cgatccagcc tccatcggct cgcatctctc cttcacgcgc ccgccgccct
     1141 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     1201 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     1261 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     1321 cttgctcaac tctagttaac ggtggagggc agtgtagtct gagcagtact cgttgctgcc
     1381 gcgcgcgcca ccagacataa tagctgacag actaacagac tgttcctttc catgggtctt
     1441 ttctgcagtc accgtcgtcg acacgtgtga tcagatatcg cggccgctct agaccaccat
     1501 gggatggtca tgtatcatcc tttttctagt agcaactgca accggtgtac attccgcacc
     1561 agagctgctg ggaggaccta gcgtgttcct gtttccccct aagcctaagg acaccctgat
     1621 gatcagccgg accccagagg tgacatgcgt ggtggtggac gtgtcccacg aggaccccga
     1681 ggtgaagttc aactggtacg tggatggcgt ggaggtgcac aatgccaaga ccaagccaag
     1741 ggaggagcag tacaactcta cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga
     1801 ctggctgaac ggcaaggagt acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat
     1861 cgagaagacc atcagcaagg caaagggaca gccaagggag cctcaggtgt ataccctgcc
     1921 acccagccgc gacgagctga caaagaacca ggtgtccctg tactgcctgg tgaagggctt
     1981 ctatccttcc gatatcgccg tggagtggga gtctaatggc cagccagaga acaattacaa
     2041 gaccacacct ccagtgctgg actccgatgg ctctttcttt ctgtattcca agctgaccgt
     2101 ggataagagc cggtggcagc agggcaacgt gtttagctgt tccgtgatgc acgaggccct
     2161 gcacaatcac tacacacaga agtctctgag cctgtccccc ggcaagggag gaggaggcag
     2221 cggcggagga ggctccggag gcggcggctc tggcggcggc ggcagcgccc ctgaactgct
     2281 gggaggacca agcgtgttcc tgtttccacc taagcccaaa gataccctga tgatctccag
     2341 gacccctgaa gtcacctgcg tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt
     2401 caactggtac gtggacggag tcgaagtcca taacgccaag accaagccta gagaagagca
     2461 gtacaactcc acctatagag tggtgtctgt gctgacagtg ctgcatcagg attggctgaa
     2521 tggcaaggaa tataagtgta aggtgagcaa taaggccctg cccgccccta tcgagaagac
     2581 catctctaaa gccaaaggac agccaaggga gccacaggtg tatacactgc cacccagccg
     2641 ggacgagctg accaagaacc aggtgagcct gacatgtctg gtcaagggct tctaccccag
     2701 tgacattgct gtggagtggg agtccaatgg ccagcctgag aacaattata aaactacccc
     2761 accagtgctg gacagcgatg gctccttctt tctgaccagc aagctgacag tggataagtc
     2821 ccgctggcag cagggaaacg tgttttcttg tagcgtgatg catgaggccc tgcataacca
     2881 ctacacccag aagtccctgt ctctgagccc aggcaaggga ggcagcggag gcggaggcag
     2941 cggaggcctg gaggtgctgt ttcagggccc tccaaacatt accaacctct gccccttcgg
     3001 cgaggtgttt aacgccacaa gatttgcctc cgtttacgcc tggaatagaa agagaatcag
     3061 caattgtgtg gccgactact ccgtgctgta taacagcgcc tctttcagca ccttcaagtg
     3121 ctacggcgtt tccccaacaa agctgaatga cctgtgcttc accaacgtgt acgccgactc
     3181 cttcgtaatt agaggcgatg aggtgcggca gatcgcacca ggccagaccg gtaagatcgc
     3241 tgactacaac tataagctgc ctgatgattt tactggctgc gtgatcgcct ggaactctaa
     3301 caacctggat agcaaggtgg gcggcaacta caactacctg taccggctgt ttcgcaagtc
     3361 taacctgaaa cctttcgaga gagacatctc cacagagatc taccaggccg gtaacacacc
     3421 ttgtaacggg gtggaaggct tcaactgtta cttccctctg caaagctacg gcttccagcc
     3481 taccaatgga gtcggctacc agccataccg ggtggtcgtg ctgtccttcg agttactcca
     3541 cgcccccgcc accgtctgcg gtggaggagg cctggtgcca cagcagagcg gaggcctgaa
     3601 cgacatcttc gaggcccaga agatcgagtg gcacgagggc tgatgaggat ccagatctgc
     3661 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3721 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3781 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3841 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atgggtaccc aggtgctgaa
     3901 gaattgaccc ggttcctcct gggccagaaa gaagcaggca catccccttc tctgtgacac
     3961 accctgtcca cgcccctggt tcttagttcc agccccactc ataggacact catagctcag
     4021 gagggctccg ccttcaatcc cacccgctaa agtacttgga gcggtctctc cctccctcat
     4081 cagcccacca aaccaaacct agcctccaag agtgggaaga aattaaagca agataggcta
     4141 ttaagtgcag agggagagaa aatgcctcca acatgtgagg aagtaatgag agaaatcata
     4201 gaattttaag gccatgattt aaggccatca tggccttaat cttccgcttc ctcgctcact
     4261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5221 tgtctatttc gttcatccat agttgcctga ctcggggggg gggggcgctg aggtctgcct
     5281 cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa
     5341 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac
     5401 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
     5461 tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct
     5521 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     5581 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     5641 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     5701 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     5761 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     5821 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     5881 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     5941 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     6001 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     6061 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     6121 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     6181 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     6241 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     6301 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
     6361 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     6421 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc
     6481 ccccccccca ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6541 aatgtattta gaaaaata
//
