LOCUS       sequence-347365-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain containing N501Y 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-N501Y-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..67)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     167..186
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        337..716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        717..920
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     871..891
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     917..941
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2850..2873
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3495..3539
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3562..3786
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4153..4170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4324..4912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4404..4423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5454..6269)
                     /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(5454..6269)
                     /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     6177..6196
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6261..6280
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     join(6547..6558,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
       61 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
      121 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
      181 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
      241 taccgcatca gattggctat tggccattgc atacgttgta tccatatcat aatatgtaca
      301 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      361 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      421 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      481 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      541 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      601 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      661 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      721 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      781 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      841 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      901 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      961 gctgttttga cctccataga agacaccggg accgatccag cctccatcgg ctcgcatctc
     1021 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     1081 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     1141 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     1201 ctccacgctt tgcctgaccc tgcttgctca actctagtta acggtggagg gcagtgtagt
     1261 ctgagcagta ctcgttgctg ccgcgcgcgc caccagacat aatagctgac agactaacag
     1321 actgttcctt tccatgggtc ttttctgcag tcaccgtcgt cgacacgtgt gatcagatat
     1381 cgcggccgct ctagaccacc atgggatggt catgtatcat cctttttcta gtagcaactg
     1441 caaccggtgt acattccgca ccagagctgc tgggaggacc tagcgtgttc ctgtttcccc
     1501 ctaagcctaa ggacaccctg atgatcagcc ggaccccaga ggtgacatgc gtggtggtgg
     1561 acgtgtccca cgaggacccc gaggtgaagt tcaactggta cgtggatggc gtggaggtgc
     1621 acaatgccaa gaccaagcca agggaggagc agtacaactc tacctatcgc gtggtgagcg
     1681 tgctgacagt gctgcaccag gactggctga acggcaagga gtacaagtgt aaggtgtcta
     1741 ataaggccct gcctgcccca atcgagaaga ccatcagcaa ggcaaaggga cagccaaggg
     1801 agcctcaggt gtataccctg ccacccagcc gcgacgagct gacaaagaac caggtgtccc
     1861 tgtactgcct ggtgaagggc ttctatcctt ccgatatcgc cgtggagtgg gagtctaatg
     1921 gccagccaga gaacaattac aagaccacac ctccagtgct ggactccgat ggctctttct
     1981 ttctgtattc caagctgacc gtggataaga gccggtggca gcagggcaac gtgtttagct
     2041 gttccgtgat gcacgaggcc ctgcacaatc actacacaca gaagtctctg agcctgtccc
     2101 ccggcaaggg aggaggaggc agcggcggag gaggctccgg aggcggcggc tctggcggcg
     2161 gcggcagcgc ccctgaactg ctgggaggac caagcgtgtt cctgtttcca cctaagccca
     2221 aagataccct gatgatctcc aggacccctg aagtcacctg cgtggtcgtg gacgtgtctc
     2281 atgaggaccc cgaagtcaag ttcaactggt acgtggacgg agtcgaagtc cataacgcca
     2341 agaccaagcc tagagaagag cagtacaact ccacctatag agtggtgtct gtgctgacag
     2401 tgctgcatca ggattggctg aatggcaagg aatataagtg taaggtgagc aataaggccc
     2461 tgcccgcccc tatcgagaag accatctcta aagccaaagg acagccaagg gagccacagg
     2521 tgtatacact gccacccagc cgggacgagc tgaccaagaa ccaggtgagc ctgacatgtc
     2581 tggtcaaggg cttctacccc agtgacattg ctgtggagtg ggagtccaat ggccagcctg
     2641 agaacaatta taaaactacc ccaccagtgc tggacagcga tggctccttc tttctgacca
     2701 gcaagctgac agtggataag tcccgctggc agcagggaaa cgtgttttct tgtagcgtga
     2761 tgcatgaggc cctgcataac cactacaccc agaagtccct gtctctgagc ccaggcaagg
     2821 gaggcagcgg aggcggaggc agcggaggcc tggaggtgct gtttcagggc cctccaaaca
     2881 ttaccaacct ctgccccttc ggcgaggtgt ttaacgccac aagatttgcc tccgtttacg
     2941 cctggaatag aaagagaatc agcaattgtg tggccgacta ctccgtgctg tataacagcg
     3001 cctctttcag caccttcaag tgctacggcg tttccccaac aaagctgaat gacctgtgct
     3061 tcaccaacgt gtacgccgac tccttcgtaa ttagaggcga tgaggtgcgg cagatcgcac
     3121 caggccagac cggtaagatc gctgactaca actataagct gcctgatgat tttacaggct
     3181 gcgtgatcgc ctggaactct aacaacctgg atagcaaggt gggcggcaac tacaactacc
     3241 tgtaccggct gtttcgcaag tctaacctga aacctttcga gagagacatc tccacagaga
     3301 tctaccaggc cggttctaca ccttgtaacg gggtggaagg cttcaactgt tacttccctc
     3361 tgcaaagcta cggcttccag cctacctacg gagtcggcta ccagccatac cgggtggtcg
     3421 tgctgtcctt cgagttactc cacgcccccg ccaccgtctg cggtggagga ggcctggtgc
     3481 cacagcagag cggaggcctg aacgacatct tcgaggccca gaagatcgag tggcacgagg
     3541 gctgatgagg atccagatct gctgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3601 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3661 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3721 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3781 ctatgggtac ccaggtgctg aagaattgac ccggttcctc ctgggccaga aagaagcagg
     3841 cacatcccct tctctgtgac acaccctgtc cacgcccctg gttcttagtt ccagccccac
     3901 tcataggaca ctcatagctc aggagggctc cgccttcaat cccacccgct aaagtacttg
     3961 gagcggtctc tccctccctc atcagcccac caaaccaaac ctagcctcca agagtgggaa
     4021 gaaattaaag caagataggc tattaagtgc agagggagag aaaatgcctc caacatgtga
     4081 ggaagtaatg agagaaatca tagaatttta aggccatgat ttaaggccat catggcctta
     4141 atcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4201 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4261 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4321 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4741 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4981 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5041 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5101 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactcggggg
     5161 gggggggcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat
     5221 cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt
     5281 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
     5341 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc
     5401 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
     5461 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     5521 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     5581 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     5641 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     5701 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     5761 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     5821 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     5881 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     5941 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     6001 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     6061 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     6121 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     6181 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
     6241 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     6301 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     6361 tgagacacaa cgtggctttc cccccccccc cattattgaa gcatttatca gggttattgt
     6421 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6481 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6541 tataaaaata ggcgtatc
//
