LOCUS       sequence-347138-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.526 variant 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 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.1.526-S477N-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..42)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     142..161
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        312..691
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        692..895
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     846..866
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     892..916
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2825..2848
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6560..6604
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6627..6851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7218..7235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7389..7977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7469..7488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8519..9334)
                     /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(8519..9334)
                     /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     9242..9261
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9326..9345
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9612..9630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
       61 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      121 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
      181 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcagattg gctattggcc
      241 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
      301 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      361 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      421 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      481 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      541 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      601 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      661 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      721 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      781 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      841 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      901 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      961 ccgggaccga tccagcctcc atcggctcgc atctctcctt cacgcgcccg ccgccctacc
     1021 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     1081 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     1141 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     1201 gctcaactct agttaacggt ggagggcagt gtagtctgag cagtactcgt tgctgccgcg
     1261 cgcgccacca gacataatag ctgacagact aacagactgt tcctttccat gggtcttttc
     1321 tgcagtcacc gtcgtcgaca cgtgtgatca gatatcgcgg ccgctctaga ccaccatggg
     1381 atggtcatgt atcatccttt ttctagtagc aactgcaacc ggtgtacatt ccgcaccaga
     1441 gctgctggga ggacctagcg tgttcctgtt tccccctaag cctaaggaca ccctgatgat
     1501 cagccggacc ccagaggtga catgcgtggt ggtggacgtg tcccacgagg accccgaggt
     1561 gaagttcaac tggtacgtgg atggcgtgga ggtgcacaat gccaagacca agccaaggga
     1621 ggagcagtac aactctacct atcgcgtggt gagcgtgctg acagtgctgc accaggactg
     1681 gctgaacggc aaggagtaca agtgtaaggt gtctaataag gccctgcctg ccccaatcga
     1741 gaagaccatc agcaaggcaa agggacagcc aagggagcct caggtgtata ccctgccacc
     1801 cagccgcgac gagctgacaa agaaccaggt gtccctgtac tgcctggtga agggcttcta
     1861 tccttccgat atcgccgtgg agtgggagtc taatggccag ccagagaaca attacaagac
     1921 cacacctcca gtgctggact ccgatggctc tttctttctg tattccaagc tgaccgtgga
     1981 taagagccgg tggcagcagg gcaacgtgtt tagctgttcc gtgatgcacg aggccctgca
     2041 caatcactac acacagaagt ctctgagcct gtcccccggc aagggaggag gaggcagcgg
     2101 cggaggaggc tccggaggcg gcggctctgg cggcggcggc agcgcccctg aactgctggg
     2161 aggaccaagc gtgttcctgt ttccacctaa gcccaaagat accctgatga tctccaggac
     2221 ccctgaagtc acctgcgtgg tcgtggacgt gtctcatgag gaccccgaag tcaagttcaa
     2281 ctggtacgtg gacggagtcg aagtccataa cgccaagacc aagcctagag aagagcagta
     2341 caactccacc tatagagtgg tgtctgtgct gacagtgctg catcaggatt ggctgaatgg
     2401 caaggaatat aagtgtaagg tgagcaataa ggccctgccc gcccctatcg agaagaccat
     2461 ctctaaagcc aaaggacagc caagggagcc acaggtgtat acactgccac ccagccggga
     2521 cgagctgacc aagaaccagg tgagcctgac atgtctggtc aagggcttct accccagtga
     2581 cattgctgtg gagtgggagt ccaatggcca gcctgagaac aattataaaa ctaccccacc
     2641 agtgctggac agcgatggct ccttctttct gaccagcaag ctgacagtgg ataagtcccg
     2701 ctggcagcag ggaaacgtgt tttcttgtag cgtgatgcat gaggccctgc ataaccacta
     2761 cacccagaag tccctgtctc tgagcccagg caagggaggc agcggaggcg gaggcagcgg
     2821 aggcctggag gtgctgtttc agggccctca gtgcgtgaac ctgaccaccc gaacccagct
     2881 cccaccagcc tacaccaaca gctttacacg gggcgtgtac taccctgaca aggtgttcag
     2941 atctagcgtc ctgcacagca ctcaggacct cttcctgccg ttcttcagca acgtgacatg
     3001 gttccacgcc atccacgtga gcggcacaaa cggaaccaag cggtttgata accccgtcct
     3061 gccattcaat gatggagttt acttcgccag tatcgagaag agtaacatca tccggggctg
     3121 gatcttcggc accaccctgg atagcaaaac acagagcctc ctgatcgtga acaatgccac
     3181 gaacgtcgtg atcaaggtgt gcgagttcca gttttgcaat gatcctttcc tgggtgtgta
     3241 ctaccacaag aacaacaaga gctggatgga aagcgagttc agagtctaca gcagcgccaa
     3301 caactgcaca ttcgagtacg tctctcagcc ttttctgatg gaccttgagg ggaaacaagg
     3361 caacttcaag aacctgagag aattcgtgtt caagaacatc gacggctact tcaaaatcta
     3421 ctccaagcac acacccatca acctggtccg ggacctccct cagggcttca gcgccctgga
     3481 acccctggtc gacctgccca taggcatcaa cataacgcgg ttccaaaccc tgctggccct
     3541 gcatagatcc tacctgactc ctggcggcag cagcagcgga tggaccgccg gagctgcagc
     3601 ctactatgtg ggctacctgc aacctagaac cttcctgctg aagtacaacg agaacggcac
     3661 aatcacagac gccgtcgact gcgccctgga ccctctctct gagacaaagt gcaccctgaa
     3721 gtccttcacc gtggaaaagg gcatctacca gaccagcaac ttccgggtgc agcctacaga
     3781 gagcatcgtg cgatttccaa acattaccaa cctctgcccc ttcggcgagg tgtttaacgc
     3841 cacaagattt gcctccgttt acgcctggaa tagaaagaga atcagcaatt gtgtggccga
     3901 ctactccgtg ctgtataaca gcgcctcttt cagcaccttc aagtgctacg gcgtttcccc
     3961 aacaaagctg aatgacctgt gcttcaccaa cgtgtacgcc gactccttcg taattagagg
     4021 cgatgaggtg cggcagatcg caccaggcca gaccggtaag atcgctgact acaactataa
     4081 gctgcctgat gattttactg gctgcgtgat cgcctggaac tctaacaacc tggatagcaa
     4141 ggtgggcggc aactacaact acctgtaccg gctgtttcgc aagtctaacc tgaaaccttt
     4201 cgagagagac atctccacag agatctacca ggccggtaac acaccttgta acggggtgga
     4261 aggcttcaac tgttacttcc ctctgcaaag ctacggcttc cagcctacca atggagtcgg
     4321 ctaccagcca taccgggtgg tcgtgctgtc cttcgagtta ctccacgccc ccgccaccgt
     4381 ctgcggtcct aagaagtcca ccaatctggt taagaacaaa tgcgtgaact tcaacttcaa
     4441 cggcctgacc gggaccggcg tgctgaccga aagcaacaaa aagttcctcc ccttccagca
     4501 gttcggccgt gatatcgctg acaccacaga tgccgtcaga gatccacaga ccctggaaat
     4561 cctggatatt acaccctgct ccttcggagg agtttctgtg atcacccccg ggaccaatac
     4621 cagcaaccag gtggctgtgc tgtaccaagg tgttaactgc accgaggttc ctgtggccat
     4681 ccacgccgat cagctgacac ctacttggag agtgtactcc actggctcca atgtgttcca
     4741 gaccagggcc ggatgtctga tcggcgccga gcacgtgaat aacagttacg agtgcgacat
     4801 ccctatcggc gccggcatct gtgccagcta ccagacccag acaaacagcc ctgggtctgc
     4861 ttcctctgta gctagccaga gcatcatcgc ctacaccatg agcctgggcg tggagaacag
     4921 cgtggcctat tccaacaact ctatcgccat tcccaccaac tttacaatta gcgtcacaac
     4981 agagatcctg cccgtgagca tgaccaagac cagcgtggac tgtacaatgt acatctgtgg
     5041 cgacagcact gaatgcagca acctgctgct gcaatacggc tccttttgca cccaactgaa
     5101 ccgggcgctg accggaatcg ccgtggaaca ggacaaaaat acccaggagg tgttcgccca
     5161 agtgaagcag atctacaaga ccccacctat caaggacttc ggcggcttta actttagcca
     5221 gattctccct gatccttcta agcctagcaa gcggagcttt atcgaggatc tgctgttcaa
     5281 caaggtcacc ctggccgatg ccggctttat caaacagtat ggcgattgcc tgggcgacat
     5341 agccgccaga gatctgatct gcgcccagaa attcaacggc ctgacagttc tcccacctct
     5401 gctgaccgac gagatgatcg ctcagtacac ctctgccctg ctggctggca ccatcacatc
     5461 tgggtggaca tttggcgccg gcgccgccct gcagatcccc tttgccatgc agatggccta
     5521 tagattcaac ggaatcggcg tgacccagaa cgtgctgtat gaaaaccaga agctgatcgc
     5581 taaccagttc aattctgcca tcggcaagat ccaggactcc ctctcctcta ccgccagcgc
     5641 cctgggcaaa ctgcaggacg tggtgaatca gaacgcccaa gccctgaaca ccctggtgaa
     5701 gcagctcagc agcaattttg gcgccatcag ctctgtgctg aacgatatcc tgtctagact
     5761 ggaccctcca gaagccgaag tccagatcga tagactgatc acaggcagac tgcagtccct
     5821 gcaaacctac gtgacccaac agctgatcag ggccgctgaa ataagagcca gcgccaatct
     5881 cgccgctacc aagatgtccg agtgtgtgct gggacagtct aaacgcgttg acttctgcgg
     5941 caaaggctat cacctgatga gcttccccca gagcgcgccg cacggcgtgg tgttcctgca
     6001 tgtgacatac gtgcctgccc aagagaagaa tttcacaacc gcccctgcca tctgccacga
     6061 cggcaaggcc cacttcccaa gagagggcgt tttcgtttcc aatggcacac actggttcgt
     6121 gacacaaaga aacttctacg aaccccagat tatcaccacc gacaacacct tcgtgagtgg
     6181 caattgtgac gtggtcatcg gaatcgtgaa caacacagtg tacgaccctc tgcaacctga
     6241 gctggactct tttaaggaag agctggacaa gtactttaaa aaccacacca gccccgatgt
     6301 ggacctgggc gacatcagtg gcattaacgc cagcgtggtg aacatccaaa aggaaatcga
     6361 cagactgaac gaggtggcca agaacctgaa cgagtccctg atcgacctgc aggagctcgg
     6421 caaatacgag cagggatctg gatacatccc cgaggccccc agagatggcc aggcctacgt
     6481 gcggaaggac ggcgagtggg tactgctgag cacattcctg ggcagatccg gaggaggcct
     6541 ggtgccacag cagagcggag gcctgaacga catcttcgag gcccagaaga tcgagtggca
     6601 cgagggctga tgaggatcca gatctgctgt gccttctagt tgccagccat ctgttgtttg
     6661 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     6721 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     6781 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     6841 gggctctatg ggtacccagg tgctgaagaa ttgacccggt tcctcctggg ccagaaagaa
     6901 gcaggcacat ccccttctct gtgacacacc ctgtccacgc ccctggttct tagttccagc
     6961 cccactcata ggacactcat agctcaggag ggctccgcct tcaatcccac ccgctaaagt
     7021 acttggagcg gtctctccct ccctcatcag cccaccaaac caaacctagc ctccaagagt
     7081 gggaagaaat taaagcaaga taggctatta agtgcagagg gagagaaaat gcctccaaca
     7141 tgtgaggaag taatgagaga aatcatagaa ttttaaggcc atgatttaag gccatcatgg
     7201 ccttaatctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     8101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     8161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8221 gggggggggg ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc
     8281 tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg
     8341 taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg
     8401 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
     8461 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     8521 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     8581 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     8641 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     8701 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     8761 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     8821 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     8881 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     8941 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     9001 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     9061 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     9121 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     9181 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     9241 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     9301 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     9361 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     9421 gattttgaga cacaacgtgg ctttcccccc ccccccatta ttgaagcatt tatcagggtt
     9481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9541 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9601 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtt
//
