LOCUS       sequence-347368-        6606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-SD1 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 6606)
  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 6606)
  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-SD1-S477N-AVI
FEATURES             Location/Qualifiers
     source          1..6606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(119..934)
                     /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(119..934)
                     /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     842..861
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     926..945
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1212..1230
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1268..1290)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1390..1409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1560..1939
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1940..2143
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2094..2114
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2140..2164
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4073..4096
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4946..4990
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    5013..5237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5604..5621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5775..6363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5855..5874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
       61 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
      121 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
      181 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
      241 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
      301 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
      361 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
      421 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
      481 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
      541 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
      601 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
      661 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
      721 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
      781 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
      841 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
      901 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
      961 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     1021 gattttgaga cacaacgtgg ctttcccccc ccccccatta ttgaagcatt tatcagggtt
     1081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1141 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1201 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     1261 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     1321 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     1381 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     1441 cgcacagatg cgtaaggaga aaataccgca tcagattggc tattggccat tgcatacgtt
     1501 gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg
     1561 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1621 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1681 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1741 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1801 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1861 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1921 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     1981 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2041 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2101 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     2161 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     2221 cagcctccat cggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     2281 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     2341 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     2401 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctag
     2461 ttaacggtgg agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga
     2521 cataatagct gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt
     2581 cgtcgacacg tgtgatcaga tatcgcggcc gctctagacc accatgggat ggtcatgtat
     2641 catccttttt ctagtagcaa ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg
     2701 acctagcgtg ttcctgtttc cccctaagcc taaggacacc ctgatgatca gccggacccc
     2761 agaggtgaca tgcgtggtgg tggacgtgtc ccacgaggac cccgaggtga agttcaactg
     2821 gtacgtggat ggcgtggagg tgcacaatgc caagaccaag ccaagggagg agcagtacaa
     2881 ctctacctat cgcgtggtga gcgtgctgac agtgctgcac caggactggc tgaacggcaa
     2941 ggagtacaag tgtaaggtgt ctaataaggc cctgcctgcc ccaatcgaga agaccatcag
     3001 caaggcaaag ggacagccaa gggagcctca ggtgtatacc ctgccaccca gccgcgacga
     3061 gctgacaaag aaccaggtgt ccctgtactg cctggtgaag ggcttctatc cttccgatat
     3121 cgccgtggag tgggagtcta atggccagcc agagaacaat tacaagacca cacctccagt
     3181 gctggactcc gatggctctt tctttctgta ttccaagctg accgtggata agagccggtg
     3241 gcagcagggc aacgtgttta gctgttccgt gatgcacgag gccctgcaca atcactacac
     3301 acagaagtct ctgagcctgt cccccggcaa gggaggagga ggcagcggcg gaggaggctc
     3361 cggaggcggc ggctctggcg gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt
     3421 gttcctgttt ccacctaagc ccaaagatac cctgatgatc tccaggaccc ctgaagtcac
     3481 ctgcgtggtc gtggacgtgt ctcatgagga ccccgaagtc aagttcaact ggtacgtgga
     3541 cggagtcgaa gtccataacg ccaagaccaa gcctagagaa gagcagtaca actccaccta
     3601 tagagtggtg tctgtgctga cagtgctgca tcaggattgg ctgaatggca aggaatataa
     3661 gtgtaaggtg agcaataagg ccctgcccgc ccctatcgag aagaccatct ctaaagccaa
     3721 aggacagcca agggagccac aggtgtatac actgccaccc agccgggacg agctgaccaa
     3781 gaaccaggtg agcctgacat gtctggtcaa gggcttctac cccagtgaca ttgctgtgga
     3841 gtgggagtcc aatggccagc ctgagaacaa ttataaaact accccaccag tgctggacag
     3901 cgatggctcc ttctttctga ccagcaagct gacagtggat aagtcccgct ggcagcaggg
     3961 aaacgtgttt tcttgtagcg tgatgcatga ggccctgcat aaccactaca cccagaagtc
     4021 cctgtctctg agcccaggca agggaggcag cggaggcgga ggcagcggag gcctggaggt
     4081 gctgtttcag ggccctcggg tgcagcctac agagagcatc gtgcgatttc caaacattac
     4141 caacctctgc cccttcggcg aggtgtttaa cgccacaaga tttgcctccg tttacgcctg
     4201 gaatagaaag agaatcagca attgtgtggc cgactactcc gtgctgtata acagcgcctc
     4261 tttcagcacc ttcaagtgct acggcgtttc cccaacaaag ctgaatgacc tgtgcttcac
     4321 caacgtgtac gccgactcct tcgtaattag aggcgatgag gtgcggcaga tcgcaccagg
     4381 ccagaccggt aagatcgctg actacaacta taagctgcct gatgatttta ctggctgcgt
     4441 gatcgcctgg aactctaaca acctggatag caaggtgggc ggcaactaca actacctgta
     4501 ccggctgttt cgcaagtcta acctgaaacc tttcgagaga gacatctcca cagagatcta
     4561 ccaggccggt aacacacctt gtaacggggt ggaaggcttc aactgttact tccctctgca
     4621 aagctacggc ttccagccta ccaatggagt cggctaccag ccataccggg tggtcgtgct
     4681 gtccttcgag ttactccacg cccccgccac cgtctgcggt cctaagaagt ccaccaatct
     4741 ggttaagaac aaatgcgtga acttcaactt caacggcctg accgggaccg gcgtgctgac
     4801 cgaaagcaac aaaaagttcc tccccttcca gcagttcggc cgtgatatcg ctgacaccac
     4861 agatgccgtc agagatccac agaccctgga aatcctggat attacaccct gctccggagg
     4921 aggcctggtg ccacagcaga gcggaggcct gaacgacatc ttcgaggccc agaagatcga
     4981 gtggcacgag ggctgatgag gatccagatc tgctgtgcct tctagttgcc agccatctgt
     5041 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     5101 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     5161 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     5221 tgcggtgggc tctatgggta cccaggtgct gaagaattga cccggttcct cctgggccag
     5281 aaagaagcag gcacatcccc ttctctgtga cacaccctgt ccacgcccct ggttcttagt
     5341 tccagcccca ctcataggac actcatagct caggagggct ccgccttcaa tcccacccgc
     5401 taaagtactt ggagcggtct ctccctccct catcagccca ccaaaccaaa cctagcctcc
     5461 aagagtggga agaaattaaa gcaagatagg ctattaagtg cagagggaga gaaaatgcct
     5521 ccaacatgtg aggaagtaat gagagaaatc atagaatttt aaggccatga tttaaggcca
     5581 tcatggcctt aatcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5641 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5701 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5761 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5821 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5881 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5941 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6001 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6061 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6121 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6181 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6241 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6301 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6361 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6421 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6481 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6541 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6601 tgactc
//
