LOCUS       sequence-347062-        9639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.1.7 variant 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 9639)
  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 9639)
  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.1.1.7-AVI
FEATURES             Location/Qualifiers
     source          1..9639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..20
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(58..80)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     180..199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        350..729
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        730..933
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     884..904
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     930..954
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2863..2886
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6589..6633
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6656..6880
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7247..7264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7418..8006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7498..7517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8548..9363)
                     /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(8548..9363)
                     /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     9271..9290
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9355..9374
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
       61 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      241 cgtaaggaga aaataccgca tcagattggc tattggccat tgcatacgtt gtatccatat
      301 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
      361 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      421 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      481 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      541 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      601 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      661 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      721 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      781 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      841 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      901 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      961 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccat
     1021 cggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     1081 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     1141 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     1201 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctag ttaacggtgg
     1261 agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga cataatagct
     1321 gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt cgtcgacacg
     1381 tgtgatcaga tatcgcggcc gctctagacc accatgggat ggtcatgtat catccttttt
     1441 ctagtagcaa ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg acctagcgtg
     1501 ttcctgtttc cccctaagcc taaggacacc ctgatgatca gccggacccc agaggtgaca
     1561 tgcgtggtgg tggacgtgtc ccacgaggac cccgaggtga agttcaactg gtacgtggat
     1621 ggcgtggagg tgcacaatgc caagaccaag ccaagggagg agcagtacaa ctctacctat
     1681 cgcgtggtga gcgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtacaag
     1741 tgtaaggtgt ctaataaggc cctgcctgcc ccaatcgaga agaccatcag caaggcaaag
     1801 ggacagccaa gggagcctca ggtgtatacc ctgccaccca gccgcgacga gctgacaaag
     1861 aaccaggtgt ccctgtactg cctggtgaag ggcttctatc cttccgatat cgccgtggag
     1921 tgggagtcta atggccagcc agagaacaat tacaagacca cacctccagt gctggactcc
     1981 gatggctctt tctttctgta ttccaagctg accgtggata agagccggtg gcagcagggc
     2041 aacgtgttta gctgttccgt gatgcacgag gccctgcaca atcactacac acagaagtct
     2101 ctgagcctgt cccccggcaa gggaggagga ggcagcggcg gaggaggctc cggaggcggc
     2161 ggctctggcg gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt gttcctgttt
     2221 ccacctaagc ccaaagatac cctgatgatc tccaggaccc ctgaagtcac ctgcgtggtc
     2281 gtggacgtgt ctcatgagga ccccgaagtc aagttcaact ggtacgtgga cggagtcgaa
     2341 gtccataacg ccaagaccaa gcctagagaa gagcagtaca actccaccta tagagtggtg
     2401 tctgtgctga cagtgctgca tcaggattgg ctgaatggca aggaatataa gtgtaaggtg
     2461 agcaataagg ccctgcccgc ccctatcgag aagaccatct ctaaagccaa aggacagcca
     2521 agggagccac aggtgtatac actgccaccc agccgggacg agctgaccaa gaaccaggtg
     2581 agcctgacat gtctggtcaa gggcttctac cccagtgaca ttgctgtgga gtgggagtcc
     2641 aatggccagc ctgagaacaa ttataaaact accccaccag tgctggacag cgatggctcc
     2701 ttctttctga ccagcaagct gacagtggat aagtcccgct ggcagcaggg aaacgtgttt
     2761 tcttgtagcg tgatgcatga ggccctgcat aaccactaca cccagaagtc cctgtctctg
     2821 agcccaggca agggaggcag cggaggcgga ggcagcggag gcctggaggt gctgtttcag
     2881 ggccctcagt gcgtgaacct gaccacccga acccagctcc caccagccta caccaacagc
     2941 tttacacggg gcgtgtacta ccctgacaag gtgttcagat ctagcgtcct gcacagcact
     3001 caggacctct tcctgccgtt cttcagcaac gtgacatggt tccacgccat cagcggcaca
     3061 aacggaacca agcggtttga taaccccgtc ctgccattca atgatggagt ttacttcgcc
     3121 agtaccgaga agagtaacat catccggggc tggatcttcg gcaccaccct ggatagcaaa
     3181 acacagagcc tcctgatcgt gaacaatgcc acgaacgtcg tgatcaaggt gtgcgagttc
     3241 cagttttgca atgatccttt cctgggtgtg taccacaaga acaacaagag ctggatggaa
     3301 agcgagttca gagtctacag cagcgccaac aactgcacat tcgagtacgt ctctcagcct
     3361 tttctgatgg accttgaggg gaaacaaggc aacttcaaga acctgagaga attcgtgttc
     3421 aagaacatcg acggctactt caaaatctac tccaagcaca cacccatcaa cctggtccgg
     3481 gacctccctc agggcttcag cgccctggaa cccctggtcg acctgcccat aggcatcaac
     3541 ataacgcggt tccaaaccct gctggccctg catagatcct acctgactcc tggcgacagc
     3601 agcagcggat ggaccgccgg agctgcagcc tactatgtgg gctacctgca acctagaacc
     3661 ttcctgctga agtacaacga gaacggcaca atcacagacg ccgtcgactg cgccctggac
     3721 cctctctctg agacaaagtg caccctgaag tccttcaccg tggaaaaggg catctaccag
     3781 accagcaact tccgggtgca gcctacagag agcatcgtgc gatttccaaa cattaccaac
     3841 ctctgcccct tcggcgaggt gtttaacgcc acaagatttg cctccgttta cgcctggaat
     3901 agaaagagaa tcagcaattg tgtggccgac tactccgtgc tgtataacag cgcctctttc
     3961 agcaccttca agtgctacgg cgtttcccca acaaagctga atgacctgtg cttcaccaac
     4021 gtgtacgccg actccttcgt aattagaggc gatgaggtgc ggcagatcgc accaggccag
     4081 accggtaaga tcgctgacta caactataag ctgcctgatg attttacagg ctgcgtgatc
     4141 gcctggaact ctaacaacct ggatagcaag gtgggcggca actacaacta cctgtaccgg
     4201 ctgtttcgca agtctaacct gaaacctttc gagagagaca tctccacaga gatctaccag
     4261 gccggttcta caccttgtaa cggggtggaa ggcttcaact gttacttccc tctgcaaagc
     4321 tacggcttcc agcctaccta cggagtcggc taccagccat accgggtggt cgtgctgtcc
     4381 ttcgagttac tccacgcccc cgccaccgtc tgcggtccta agaagtccac caatctggtt
     4441 aagaacaaat gcgtgaactt caacttcaac ggcctgaccg ggaccggcgt gctgaccgaa
     4501 agcaacaaaa agttcctccc cttccagcag ttcggccgtg atatcgatga caccacagat
     4561 gccgtcagag atccacagac cctggaaatc ctggatatta caccctgctc cttcggagga
     4621 gtttctgtga tcacccccgg gaccaatacc agcaaccagg tggctgtgct gtaccaaggt
     4681 gttaactgca ccgaggttcc tgtggccatc cacgccgatc agctgacacc tacttggaga
     4741 gtgtactcca ctggctccaa tgtgttccag accagggccg gatgtctgat cggcgccgag
     4801 cacgtgaata acagttacga gtgcgacatc cctatcggcg ccggcatctg tgccagctac
     4861 cagacccaga caaacagcca cgggtctgct tcctctgtag ctagccagag catcatcgcc
     4921 tacaccatga gcctgggcgc agagaacagc gtggcctatt ccaacaactc tatcgccatt
     4981 cccatcaact ttacaattag cgtcacaaca gagatcctgc ccgtgagcat gaccaagacc
     5041 agcgtggact gtacaatgta catctgtggc gacagcactg aatgcagcaa cctgctgctg
     5101 caatacggct ccttttgcac ccaactgaac cgggcgctga ccggaatcgc cgtggaacag
     5161 gacaaaaata cccaggaggt gttcgcccaa gtgaagcaga tctacaagac cccacctatc
     5221 aaggacttcg gcggctttaa ctttagccag attctccctg atccttctaa gcctagcaag
     5281 cggagcttta tcgaggatct gctgttcaac aaggtcaccc tggccgatgc cggctttatc
     5341 aaacagtatg gcgattgcct gggcgacata gccgccagag atctgatctg cgcccagaaa
     5401 ttcaacggcc tgacagttct cccacctctg ctgaccgacg agatgatcgc tcagtacacc
     5461 tctgccctgc tggctggcac catcacatct gggtggacat ttggcgccgg cgccgccctg
     5521 cagatcccct ttgccatgca gatggcctat agattcaacg gaatcggcgt gacccagaac
     5581 gtgctgtatg aaaaccagaa gctgatcgct aaccagttca attctgccat cggcaagatc
     5641 caggactccc tctcctctac cgccagcgcc ctgggcaaac tgcaggacgt ggtgaatcag
     5701 aacgcccaag ccctgaacac cctggtgaag cagctcagca gcaattttgg cgccatcagc
     5761 tctgtgctga acgatatcct ggctagactg gaccctccag aagccgaagt ccagatcgat
     5821 agactgatca caggtagact gcagtccctg caaacctacg tgacccaaca gctgatcagg
     5881 gccgctgaaa taagagccag cgccaatctc gccgctacca agatgtccga gtgtgtgctg
     5941 ggacagtcta aacgcgttga cttctgcggc aaaggctatc acctgatgag cttcccccag
     6001 agcgcgccgc acggcgtggt gttcctgcat gtgacatacg tgcctgccca agagaagaat
     6061 ttcacaaccg cccctgccat ctgccacgac ggcaaggccc acttcccaag agagggcgtt
     6121 ttcgtttcca atggcacaca ctggttcgtg acacaaagaa acttctacga accccagatt
     6181 atcaccaccc acaacacctt cgtgagtggc aattgtgacg tggtcatcgg aatcgtgaac
     6241 aacacagtgt acgaccctct gcaacctgag ctggactctt ttaaggaaga gctggacaag
     6301 tactttaaaa accacaccag ccccgatgtg gacctgggcg acatcagtgg cattaacgcc
     6361 agcgtggtga acatccaaaa ggaaatcgac agactgaacg aggtggccaa gaacctgaac
     6421 gagtccctga tcgacctgca ggagctcggc aaatacgagc agggatctgg atacatcccc
     6481 gaggccccca gagatggcca ggcctacgtg cggaaggacg gcgagtgggt actgctgagc
     6541 acattcctgg gcagatccgg aggaggcctg gtgccacagc agagcggagg cctgaacgac
     6601 atcttcgagg cccagaagat cgagtggcac gagggctgat gaggatccag atctgctgtg
     6661 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6721 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6781 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6841 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gtacccaggt gctgaagaat
     6901 tgacccggtt cctcctgggc cagaaagaag caggcacatc cccttctctg tgacacaccc
     6961 tgtccacgcc cctggttctt agttccagcc ccactcatag gacactcata gctcaggagg
     7021 gctccgcctt caatcccacc cgctaaagta cttggagcgg tctctccctc cctcatcagc
     7081 ccaccaaacc aaacctagcc tccaagagtg ggaagaaatt aaagcaagat aggctattaa
     7141 gtgcagaggg agagaaaatg cctccaacat gtgaggaagt aatgagagaa atcatagaat
     7201 tttaaggcca tgatttaagg ccatcatggc cttaatcttc cgcttcctcg ctcactgact
     7261 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7321 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8221 tatttcgttc atccatagtt gcctgactcg gggggggggg gcgctgaggt ctgcctcgtg
     8281 aagaaggtgt tgctgactca taccaggcct gaatcgcccc atcatccagc cagaaagtga
     8341 gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt ttgaactttt
     8401 gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag
     8461 caaaagttcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca
     8521 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     8581 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     8641 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     8701 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     8761 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     8821 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     8881 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     8941 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     9001 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     9061 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     9121 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     9181 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     9241 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     9301 agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct
     9361 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     9421 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttccccccc
     9481 cccccattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9601 cgtctaagaa accattatta tcatgacatt aacctataa
//
