LOCUS       sequence-347141-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.617 variant containing
            G142D-E154K-L452R-E484Q-D614G-P681R-Q1071H-H1101D 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 9648)
  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 9648)
  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-S2P-B.1.617-G142D-E154K-L452R-E484Q-D614G-P681R-
            Q1071H-H1101D-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..54)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     154..173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        324..703
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        704..907
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     858..878
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     904..928
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2837..2860
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6572..6616
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6639..6863
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7230..7247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7401..7989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7481..7500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8531..9346)
                     /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(8531..9346)
                     /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     9254..9273
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9338..9357
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9624..9642
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
       61 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
      121 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
      181 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagat
      241 tggctattgg ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc
      301 atgtccaaca ttaccgccat gttgacattg attattgact agttattaat agtaatcaat
      361 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      421 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      481 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      541 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      601 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      661 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      721 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      781 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      841 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      901 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      961 ccatagaaga caccgggacc gatccagcct ccatcggctc gcatctctcc ttcacgcgcc
     1021 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     1081 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     1141 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     1201 ctgaccctgc ttgctcaact ctagttaacg gtggagggca gtgtagtctg agcagtactc
     1261 gttgctgccg cgcgcgccac cagacataat agctgacaga ctaacagact gttcctttcc
     1321 atgggtcttt tctgcagtca ccgtcgtcga cacgtgtgat cagatatcgc ggccgctcta
     1381 gaccaccatg ggatggtcat gtatcatcct ttttctagta gcaactgcaa ccggtgtaca
     1441 ttccgcacca gagctgctgg gaggacctag cgtgttcctg tttcccccta agcctaagga
     1501 caccctgatg atcagccgga ccccagaggt gacatgcgtg gtggtggacg tgtcccacga
     1561 ggaccccgag gtgaagttca actggtacgt ggatggcgtg gaggtgcaca atgccaagac
     1621 caagccaagg gaggagcagt acaactctac ctatcgcgtg gtgagcgtgc tgacagtgct
     1681 gcaccaggac tggctgaacg gcaaggagta caagtgtaag gtgtctaata aggccctgcc
     1741 tgccccaatc gagaagacca tcagcaaggc aaagggacag ccaagggagc ctcaggtgta
     1801 taccctgcca cccagccgcg acgagctgac aaagaaccag gtgtccctgt actgcctggt
     1861 gaagggcttc tatccttccg atatcgccgt ggagtgggag tctaatggcc agccagagaa
     1921 caattacaag accacacctc cagtgctgga ctccgatggc tctttctttc tgtattccaa
     1981 gctgaccgtg gataagagcc ggtggcagca gggcaacgtg tttagctgtt ccgtgatgca
     2041 cgaggccctg cacaatcact acacacagaa gtctctgagc ctgtcccccg gcaagggagg
     2101 aggaggcagc ggcggaggag gctccggagg cggcggctct ggcggcggcg gcagcgcccc
     2161 tgaactgctg ggaggaccaa gcgtgttcct gtttccacct aagcccaaag ataccctgat
     2221 gatctccagg acccctgaag tcacctgcgt ggtcgtggac gtgtctcatg aggaccccga
     2281 agtcaagttc aactggtacg tggacggagt cgaagtccat aacgccaaga ccaagcctag
     2341 agaagagcag tacaactcca cctatagagt ggtgtctgtg ctgacagtgc tgcatcagga
     2401 ttggctgaat ggcaaggaat ataagtgtaa ggtgagcaat aaggccctgc ccgcccctat
     2461 cgagaagacc atctctaaag ccaaaggaca gccaagggag ccacaggtgt atacactgcc
     2521 acccagccgg gacgagctga ccaagaacca ggtgagcctg acatgtctgg tcaagggctt
     2581 ctaccccagt gacattgctg tggagtggga gtccaatggc cagcctgaga acaattataa
     2641 aactacccca ccagtgctgg acagcgatgg ctccttcttt ctgaccagca agctgacagt
     2701 ggataagtcc cgctggcagc agggaaacgt gttttcttgt agcgtgatgc atgaggccct
     2761 gcataaccac tacacccaga agtccctgtc tctgagccca ggcaagggag gcagcggagg
     2821 cggaggcagc ggaggcctgg aggtgctgtt tcagggccct cagtgcgtga acctgaccac
     2881 ccgaacccag ctcccaccag cctacaccaa cagctttaca cggggcgtgt actaccctga
     2941 caaggtgttc agatctagcg tcctgcacag cactcaggac ctcttcctgc cgttcttcag
     3001 caacgtgaca tggttccacg ccatccacgt gagcggcaca aacggaacca agcggtttga
     3061 taaccccgtc ctgccattca atgatggagt ttacttcgcc agtaccgaga agagtaacat
     3121 catccggggc tggatcttcg gcaccaccct ggatagcaaa acacagagcc tcctgatcgt
     3181 gaacaatgcc acgaacgtcg tgatcaaggt gtgcgagttc cagttttgca atgatccttt
     3241 cctggacgtg tactaccaca agaacaacaa gagctggatg aagagcgagt tcagagtcta
     3301 cagcagcgcc aacaactgca cattcgagta cgtctctcag ccttttctga tggaccttga
     3361 ggggaaacaa ggcaacttca agaacctgag agaattcgtg ttcaagaaca tcgacggcta
     3421 cttcaaaatc tactccaagc acacacccat caacctggtc cgggacctcc ctcagggctt
     3481 cagcgccctg gaacccctgg tcgacctgcc cataggcatc aacataacgc ggttccaaac
     3541 cctgctggcc ctgcatagat cctacctgac tcctggcgac agcagcagcg gatggaccgc
     3601 cggagctgca gcctactatg tgggctacct gcaacctaga accttcctgc tgaagtacaa
     3661 cgagaacggc acaatcacag acgccgtcga ctgcgccctg gaccctctct ctgagacaaa
     3721 gtgcaccctg aagtccttca ccgtggaaaa gggcatctac cagaccagca acttccgggt
     3781 gcagcctaca gagagcatcg tgcgatttcc aaacattacc aacctctgcc ccttcggcga
     3841 ggtgtttaac gccacaagat ttgcctccgt ttacgcctgg aatagaaaga gaatcagcaa
     3901 ttgtgtggcc gactactccg tgctgtataa cagcgcctct ttcagcacct tcaagtgcta
     3961 cggcgtttcc ccaacaaagc tgaatgacct gtgcttcacc aacgtgtacg ccgactcctt
     4021 cgtaattaga ggcgatgagg tgcggcagat cgcaccaggc cagaccggta agatcgctga
     4081 ctacaactat aagctgcctg atgattttac aggctgcgtg atcgcctgga actctaacaa
     4141 cctggatagc aaggtgggcg gcaactacaa ctacagatac cggctgtttc gcaagtctaa
     4201 cctgaaacct ttcgagagag acatctccac agagatctac caggccggtt ctacaccttg
     4261 taacggggtg cagggcttca actgttactt ccctctgcaa agctacggct tccagcctac
     4321 caatggagtc ggctaccagc cataccgggt ggtcgtgctg tccttcgagt tactccacgc
     4381 ccccgccacc gtctgcggtc ctaagaagtc caccaatctg gttaagaaca aatgcgtgaa
     4441 cttcaacttc aacggcctga ccgggaccgg cgtgctgacc gaaagcaaca aaaagttcct
     4501 ccccttccag cagttcggcc gtgatatcgc tgacaccaca gatgccgtca gagatccaca
     4561 gaccctggaa atcctggata ttacaccctg ctccttcgga ggagtttctg tgatcacccc
     4621 cgggaccaat accagcaacc aggtggctgt gctgtaccaa ggtgttaact gcaccgaggt
     4681 tcctgtggcc atccacgccg atcagctgac acctacttgg agagtgtact ccactggctc
     4741 caatgtgttc cagaccaggg ccggatgtct gatcggcgcc gagcacgtga ataacagtta
     4801 cgagtgcgac atccctatcg gcgccggcat ctgtgccagc taccagaccc agacaaacag
     4861 cagagggtct gcttcctctg tagctagcca gagcatcatc gcctacacca tgagcctggg
     4921 cgcagagaac agcgtggcct attccaacaa ctctatcgcc attcccacca actttacaat
     4981 tagcgtcaca acagagatcc tgcccgtgag catgaccaag accagcgtgg actgtacaat
     5041 gtacatctgt ggcgacagca ctgaatgcag caacctgctg ctgcaatacg gctccttttg
     5101 cacccaactg aaccgggcgc tgaccggaat cgccgtggaa caggacaaaa atacccagga
     5161 ggtgttcgcc caagtgaagc agatctacaa gaccccacct atcaaggact tcggcggctt
     5221 taactttagc cagattctcc ctgatccttc taagcctagc aagcggagct ttatcgagga
     5281 tctgctgttc aacaaggtca ccctggccga tgccggcttt atcaaacagt atggcgattg
     5341 cctgggcgac atagccgcca gagatctgat ctgcgcccag aaattcaacg gcctgacagt
     5401 tctcccacct ctgctgaccg acgagatgat cgctcagtac acctctgccc tgctggctgg
     5461 caccatcaca tctgggtgga catttggcgc cggcgccgcc ctgcagatcc cctttgccat
     5521 gcagatggcc tatagattca acggaatcgg cgtgacccag aacgtgctgt atgaaaacca
     5581 gaagctgatc gctaaccagt tcaattctgc catcggcaag atccaggact ccctctcctc
     5641 taccgccagc gccctgggca aactgcagga cgtggtgaat cagaacgccc aagccctgaa
     5701 caccctggtg aagcagctca gcagcaattt tggcgccatc agctctgtgc tgaacgatat
     5761 cctgtctaga ctggaccctc cagaagccga agtccagatc gatagactga tcacaggcag
     5821 actgcagtcc ctgcaaacct acgtgaccca acagctgatc agggccgctg aaataagagc
     5881 cagcgccaat ctcgccgcta ccaagatgtc cgagtgtgtg ctgggacagt ctaaacgcgt
     5941 tgacttctgc ggcaaaggct atcacctgat gagcttcccc cagagcgcgc cgcacggcgt
     6001 ggtgttcctg catgtgacat acgtgcctgc ccacgagaag aatttcacaa ccgcccctgc
     6061 catctgccac gacggcaagg cccacttccc aagagagggc gttttcgttt ccaatggcac
     6121 agactggttc gtgacacaaa gaaacttcta cgaaccccag attatcacca ccgacaacac
     6181 cttcgtgagt ggcaattgtg acgtggtcat cggaatcgtg aacaacacag tgtacgaccc
     6241 tctgcaacct gagctggact cttttaagga agagctggac aagtacttta aaaaccacac
     6301 cagccccgat gtggacctgg gcgacatcag tggcattaac gccagcgtgg tgaacatcca
     6361 aaaggaaatc gacagactga acgaggtggc caagaacctg aacgagtccc tgatcgacct
     6421 gcaggagctc ggcaaatacg agcagggatc tggatacatc cccgaggccc ccagagatgg
     6481 ccaggcctac gtgcggaagg acggcgagtg ggtactgctg agcacattcc tgggcagatc
     6541 cggaggaggc ctggtgccac agcagagcgg aggcctgaac gacatcttcg aggcccagaa
     6601 gatcgagtgg cacgagggct gatgaggatc cagatctgct gtgccttcta gttgccagcc
     6661 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     6721 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     6781 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     6841 tggggatgcg gtgggctcta tgggtaccca ggtgctgaag aattgacccg gttcctcctg
     6901 ggccagaaag aagcaggcac atccccttct ctgtgacaca ccctgtccac gcccctggtt
     6961 cttagttcca gccccactca taggacactc atagctcagg agggctccgc cttcaatccc
     7021 acccgctaaa gtacttggag cggtctctcc ctccctcatc agcccaccaa accaaaccta
     7081 gcctccaaga gtgggaagaa attaaagcaa gataggctat taagtgcaga gggagagaaa
     7141 atgcctccaa catgtgagga agtaatgaga gaaatcatag aattttaagg ccatgattta
     7201 aggccatcat ggccttaatc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7261 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7321 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7381 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7441 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7501 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7561 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7621 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7681 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7741 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7801 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7861 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7921 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7981 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8041 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8101 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8161 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8221 gttgcctgac tcgggggggg ggggcgctga ggtctgcctc gtgaagaagg tgttgctgac
     8281 tcataccagg cctgaatcgc cccatcatcc agccagaaag tgagggagcc acggttgatg
     8341 agagctttgt tgtaggtgga ccagttggtg attttgaact tttgctttgc cacggaacgg
     8401 tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcgatttatt
     8461 caacaaagcc gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa
     8521 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     8581 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     8641 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     8701 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     8761 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt
     8821 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     8881 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     8941 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     9001 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     9061 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     9121 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     9181 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     9241 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     9301 ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     9361 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     9421 tgtaacatca gagattttga gacacaacgt ggctttcccc ccccccccat tattgaagca
     9481 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9541 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     9601 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttc
//
