LOCUS       sequence-385180-        9633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the BQ.1.1 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 9633)
  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 9633)
  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-BQ.1.1-AVI
FEATURES             Location/Qualifiers
     source          1..9633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        321..700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        701..904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     855..875
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     901..925
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2834..2857
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6554..6598
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6621..6845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7212..7229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7383..7971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7463..7482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8513..9328)
                     /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(8513..9328)
                     /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     9236..9255
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9320..9339
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9606..9624
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcagattgg
      241 ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
      301 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
      361 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      421 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      721 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      781 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      841 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      901 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
      961 tagaagacac cgggaccgat ccagcctcca tcggctcgca tctctccttc acgcgcccgc
     1021 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     1081 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     1141 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     1201 accctgcttg ctcaactcta gttaacggtg gagggcagtg tagtctgagc agtactcgtt
     1261 gctgccgcgc gcgccaccag acataatagc tgacagacta acagactgtt cctttccatg
     1321 ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag atatcgcggc cgctctagac
     1381 caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg gtgtacattc
     1441 cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc ctaaggacac
     1501 cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt cccacgagga
     1561 ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg ccaagaccaa
     1621 gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga cagtgctgca
     1681 ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg ccctgcctgc
     1741 cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc aggtgtatac
     1801 cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact gcctggtgaa
     1861 gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc cagagaacaa
     1921 ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt attccaagct
     1981 gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg tgatgcacga
     2041 ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca agggaggagg
     2101 aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca gcgcccctga
     2161 actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata ccctgatgat
     2221 ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg accccgaagt
     2281 caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca agcctagaga
     2341 agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc atcaggattg
     2401 gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg cccctatcga
     2461 gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata cactgccacc
     2521 cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca agggcttcta
     2581 ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca attataaaac
     2641 taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc tgacagtgga
     2701 taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg aggccctgca
     2761 taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca gcggaggcgg
     2821 aggcagcgga ggcctggagg tgctgtttca gggccctcag tgcgtgaacc tgatcacccg
     2881 aacccagtcc tacaccaaca gctttacacg gggcgtgtac taccctgaca aggtgttcag
     2941 atctagcgtc ctgcacagca ctcaggacct cttcctgccg ttcttcagca acgtgacatg
     3001 gttccacgcc atcagcggca caaacggaac caagcggttt gataaccccg tcctgccatt
     3061 caatgatgga gtttacttcg ccagtaccga gaagagtaac atcatccggg gctggatctt
     3121 cggcaccacc ctggatagca aaacacagag cctcctgatc gtgaacaatg ccacgaacgt
     3181 cgtgatcaag gtgtgcgagt tccagttttg caatgatcct ttcctggatg tgtactacca
     3241 caagaacaac aagagctgga tggaaagcga gttcagagtc tacagcagcg ccaacaactg
     3301 cacattcgag tacgtctctc agccttttct gatggacctt gaggggaaac aaggcaactt
     3361 caagaacctg agagaattcg tgttcaagaa catcgacggc tacttcaaaa tctactccaa
     3421 gcacacaccc atcaacctgg gccgggacct ccctcagggc ttcagcgccc tggaacccct
     3481 ggtcgacctg cccataggca tcaacataac gcggttccaa accctgctgg ccctgcatag
     3541 atcctacctg actcctggcg acagcagcag cggatggacc gccggagctg cagcctacta
     3601 tgtgggctac ctgcaaccta gaaccttcct gctgaagtac aacgagaacg gcacaatcac
     3661 agacgccgtc gactgcgccc tggaccctct ctctgagaca aagtgcaccc tgaagtcctt
     3721 caccgtggaa aagggcatct accagaccag caacttccgg gtgcagccta cagagagcat
     3781 cgtgcgattt ccaaacatta ccaacctctg ccccttcgac gaggtgttta acgccacaac
     3841 ctttgcctcc gtttacgcct ggaatagaaa gagaatcagc aattgtgtgg ccgactactc
     3901 cgtgctgtat aacttcgccc ctttcttcgc cttcaagtgc tacggcgttt ccccaacaaa
     3961 gctgaatgac ctgtgcttca ccaacgtgta cgccgactcc ttcgtaatta gaggcaatga
     4021 ggtgtcccag atcgcaccag gccagaccgg taacatcgct gactacaact ataagctgcc
     4081 tgatgatttt acaggctgcg tgatcgcctg gaactccaac aagctggata gcaccgtggg
     4141 cggcaactac aactacagat accggctgtt tcgcaagtct aagctgaaac ctttcgagag
     4201 agacatctcc acagagatct accaggccgg taacaagcct tgtaacgggg tggccggcgt
     4261 gaactgttac ttccctctgc aaagctacgg cttccggcct acctatggag tcggccacca
     4321 gccataccgg gtggtcgtgc tgtccttcga gttactccac gcccccgcca ccgtctgcgg
     4381 tcctaagaag tccaccaatc tggttaagaa caaatgcgtg aacttcaact tcaacggcct
     4441 gaccgggacc ggcgtgctga ccgaaagcaa caaaaagttc ctccccttcc agcagttcgg
     4501 ccgtgatatc gctgacacca cagatgccgt cagagatcca cagaccctgg aaatcctgga
     4561 tattacaccc tgctccttcg gaggagtttc tgtgatcacc cccgggacca ataccagcaa
     4621 ccaggtggct gtgctgtacc aaggtgttaa ctgcaccgag gttcctgtgg ccatccacgc
     4681 cgatcagctg acacctactt ggagagtgta ctccactggc tccaatgtgt tccagaccag
     4741 ggccggatgt ctgatcggcg ccgagtacgt gaataacagt tacgagtgcg acatccctat
     4801 cggcgccggc atctgtgcca gctaccagac ccagacaaag agccatggca gcgcctcctc
     4861 tgtagctagc cagagcatca tcgcctacac catgagcctg ggcgcagaga acagcgtggc
     4921 ctattccaac aactctatcg ccattcccac caactttaca attagcgtca caacagagat
     4981 cctgcccgtg agcatgacca agaccagcgt ggactgtaca atgtacatct gtggcgacag
     5041 cactgaatgc agcaacctgc tgctgcaata cggctccttt tgcacccaac tgaagcgggc
     5101 gctgaccgga atcgccgtgg aacaggacaa aaatacccag gaggtgttcg cccaagtgaa
     5161 gcagatctac aagaccccac ctatcaagta cttcggcggc tttaacttta gccagattct
     5221 ccctgatcct tctaagccta gcaagcggag ctttatcgag gatctgctgt tcaacaaggt
     5281 caccctggcc gatgccggct ttatcaaaca gtatggcgat tgtctgggcg acatagccgc
     5341 cagagatctg atctgcgccc agaaattcaa cggcctgaca gttctcccac ctctgctgac
     5401 cgacgagatg atcgctcagt acacctctgc cctgctggct ggcaccatca catctgggtg
     5461 gacatttggc gccggcgccg ccctgcagat cccctttgcc atgcagatgg cctatagatt
     5521 caacggaatc ggcgtgaccc agaacgtgct gtatgaaaac cagaagctga tcgctaacca
     5581 gttcaattct gccatcggca agatccagga ctccctctcc tctaccgcca gcgccctggg
     5641 caaactgcag gacgtggtga atcacaacgc ccaagccctg aacaccctgg tgaagcagct
     5701 cagcagcaag tttggcgcca tcagctctgt gctgaacgat atcctgtcta gactggaccc
     5761 ccctgaagcc gaagtccaga tcgatagact gatcacaggt agactgcagt ccctgcaaac
     5821 ctacgtgacc caacagctga tcagggccgc tgaaataaga gccagcgcca atctcgccgc
     5881 taccaagatg tccgagtgtg tgctgggaca gtctaaacgc gttgacttct gcggcaaagg
     5941 ctatcacctg atgagcttcc cccagagcgc gccgcacggc gtggtgttcc tgcatgtgac
     6001 atacgtgcct gcccaagaga agaatttcac aaccgcccct gccatctgcc acgacggcaa
     6061 ggcccacttc ccaagagagg gcgttttcgt ttccaatggc acacactggt tcgtgacaca
     6121 aagaaacttc tacgaacccc agattatcac caccgacaac accttcgtga gtggcaattg
     6181 tgacgtggtc atcggaatcg tgaacaacac agtgtacgac cctctgcaac ctgagctgga
     6241 ctcttttaag gaagagctgg acaagtactt taaaaaccac accagccccg atgtggacct
     6301 gggcgacatc agtggcatta acgccagcgt ggtgaacatc caaaaggaaa tcgacagact
     6361 gaacgaggtg gccaagaacc tgaacgagtc cctgatcgac ctgcaggagc tcggcaaata
     6421 cgagcaggga tctggataca tccccgaggc ccccagagat ggccaggcct acgtgcggaa
     6481 ggacggcgag tgggtactgc tgagcacatt cctgggcaga tccggaggag gcctggtgcc
     6541 acagcagagc ggaggcctga acgacatctt cgaggcccag aagatcgagt ggcacgaggg
     6601 ctgatgagga tccagatctg ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6661 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     6721 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     6781 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     6841 tatgggtacc caggtgctga agaattgacc cggttcctcc tgggccagaa agaagcaggc
     6901 acatcccctt ctctgtgaca caccctgtcc acgcccctgg ttcttagttc cagccccact
     6961 cataggacac tcatagctca ggagggctcc gccttcaatc ccacccgcta aagtacttgg
     7021 agcggtctct ccctccctca tcagcccacc aaaccaaacc tagcctccaa gagtgggaag
     7081 aaattaaagc aagataggct attaagtgca gagggagaga aaatgcctcc aacatgtgag
     7141 gaagtaatga gagaaatcat agaattttaa ggccatgatt taaggccatc atggccttaa
     7201 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7261 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7321 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7381 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7441 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7501 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7561 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7621 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7681 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7741 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7801 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7861 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7921 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7981 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8041 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8101 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8161 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actcgggggg
     8221 ggggggcgct gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc
     8281 gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg
     8341 gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga
     8401 tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccgccgtccc
     8461 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     8521 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     8581 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     8641 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     8701 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     8761 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     8821 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     8881 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     8941 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     9001 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     9061 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     9121 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     9181 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     9241 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
     9301 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     9361 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     9421 gagacacaac gtggctttcc cccccccccc attattgaag catttatcag ggttattgtc
     9481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9541 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     9601 ataaaaatag gcgtatcacg aggccctttc gtc
//
