LOCUS       sequence-347139-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.526 variant containing
            E484K 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-E484K-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     120..139
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        290..669
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        670..873
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     824..844
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     870..894
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2803..2826
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6538..6582
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6605..6829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7196..7213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7367..7955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7447..7466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8497..9312)
                     /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(8497..9312)
                     /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     9220..9239
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9304..9323
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9590..9608
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(9646..9648,1..20))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
       61 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      121 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      181 cgtaaggaga aaataccgca tcagattggc tattggccat tgcatacgtt gtatccatat
      241 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
      301 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      361 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      421 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      481 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      541 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      601 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      661 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      721 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      781 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      841 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      901 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccat
      961 cggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     1021 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     1081 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     1141 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctag ttaacggtgg
     1201 agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga cataatagct
     1261 gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt cgtcgacacg
     1321 tgtgatcaga tatcgcggcc gctctagacc accatgggat ggtcatgtat catccttttt
     1381 ctagtagcaa ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg acctagcgtg
     1441 ttcctgtttc cccctaagcc taaggacacc ctgatgatca gccggacccc agaggtgaca
     1501 tgcgtggtgg tggacgtgtc ccacgaggac cccgaggtga agttcaactg gtacgtggat
     1561 ggcgtggagg tgcacaatgc caagaccaag ccaagggagg agcagtacaa ctctacctat
     1621 cgcgtggtga gcgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtacaag
     1681 tgtaaggtgt ctaataaggc cctgcctgcc ccaatcgaga agaccatcag caaggcaaag
     1741 ggacagccaa gggagcctca ggtgtatacc ctgccaccca gccgcgacga gctgacaaag
     1801 aaccaggtgt ccctgtactg cctggtgaag ggcttctatc cttccgatat cgccgtggag
     1861 tgggagtcta atggccagcc agagaacaat tacaagacca cacctccagt gctggactcc
     1921 gatggctctt tctttctgta ttccaagctg accgtggata agagccggtg gcagcagggc
     1981 aacgtgttta gctgttccgt gatgcacgag gccctgcaca atcactacac acagaagtct
     2041 ctgagcctgt cccccggcaa gggaggagga ggcagcggcg gaggaggctc cggaggcggc
     2101 ggctctggcg gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt gttcctgttt
     2161 ccacctaagc ccaaagatac cctgatgatc tccaggaccc ctgaagtcac ctgcgtggtc
     2221 gtggacgtgt ctcatgagga ccccgaagtc aagttcaact ggtacgtgga cggagtcgaa
     2281 gtccataacg ccaagaccaa gcctagagaa gagcagtaca actccaccta tagagtggtg
     2341 tctgtgctga cagtgctgca tcaggattgg ctgaatggca aggaatataa gtgtaaggtg
     2401 agcaataagg ccctgcccgc ccctatcgag aagaccatct ctaaagccaa aggacagcca
     2461 agggagccac aggtgtatac actgccaccc agccgggacg agctgaccaa gaaccaggtg
     2521 agcctgacat gtctggtcaa gggcttctac cccagtgaca ttgctgtgga gtgggagtcc
     2581 aatggccagc ctgagaacaa ttataaaact accccaccag tgctggacag cgatggctcc
     2641 ttctttctga ccagcaagct gacagtggat aagtcccgct ggcagcaggg aaacgtgttt
     2701 tcttgtagcg tgatgcatga ggccctgcat aaccactaca cccagaagtc cctgtctctg
     2761 agcccaggca agggaggcag cggaggcgga ggcagcggag gcctggaggt gctgtttcag
     2821 ggccctcagt gcgtgaacct gaccacccga acccagctcc caccagccta caccaacagc
     2881 tttacacggg gcgtgtacta ccctgacaag gtgttcagat ctagcgtcct gcacagcact
     2941 caggacctct tcctgccgtt cttcagcaac gtgacatggt tccacgccat ccacgtgagc
     3001 ggcacaaacg gaaccaagcg gtttgataac cccgtcctgc cattcaatga tggagtttac
     3061 ttcgccagta tcgagaagag taacatcatc cggggctgga tcttcggcac caccctggat
     3121 agcaaaacac agagcctcct gatcgtgaac aatgccacga acgtcgtgat caaggtgtgc
     3181 gagttccagt tttgcaatga tcctttcctg ggtgtgtact accacaagaa caacaagagc
     3241 tggatggaaa gcgagttcag agtctacagc agcgccaaca actgcacatt cgagtacgtc
     3301 tctcagcctt ttctgatgga ccttgagggg aaacaaggca acttcaagaa cctgagagaa
     3361 ttcgtgttca agaacatcga cggctacttc aaaatctact ccaagcacac acccatcaac
     3421 ctggtccggg acctccctca gggcttcagc gccctggaac ccctggtcga cctgcccata
     3481 ggcatcaaca taacgcggtt ccaaaccctg ctggccctgc atagatccta cctgactcct
     3541 ggcggcagca gcagcggatg gaccgccgga gctgcagcct actatgtggg ctacctgcaa
     3601 cctagaacct tcctgctgaa gtacaacgag aacggcacaa tcacagacgc cgtcgactgc
     3661 gccctggacc ctctctctga gacaaagtgc accctgaagt ccttcaccgt ggaaaagggc
     3721 atctaccaga ccagcaactt ccgggtgcag cctacagaga gcatcgtgcg atttccaaac
     3781 attaccaacc tctgcccctt cggcgaggtg tttaacgcca caagatttgc ctccgtttac
     3841 gcctggaata gaaagagaat cagcaattgt gtggccgact actccgtgct gtataacagc
     3901 gcctctttca gcaccttcaa gtgctacggc gtttccccaa caaagctgaa tgacctgtgc
     3961 ttcaccaacg tgtacgccga ctccttcgta attagaggcg atgaggtgcg gcagatcgca
     4021 ccaggccaga ccggtaagat cgctgactac aactataagc tgcctgatga ttttacaggc
     4081 tgcgtgatcg cctggaactc taacaacctg gatagcaagg tgggcggcaa ctacaactac
     4141 ctgtaccggc tgtttcgcaa gtctaacctg aaacctttcg agagagacat ctccacagag
     4201 atctaccagg ccggttctac accttgtaac ggggtgaaag gcttcaactg ttacttccct
     4261 ctgcaaagct acggcttcca gcctaccaat ggagtcggct accagccata ccgggtggtc
     4321 gtgctgtcct tcgagttact ccacgccccc gccaccgtct gcggtcctaa gaagtccacc
     4381 aatctggtta agaacaaatg cgtgaacttc aacttcaacg gcctgaccgg gaccggcgtg
     4441 ctgaccgaaa gcaacaaaaa gttcctcccc ttccagcagt tcggccgtga tatcgctgac
     4501 accacagatg ccgtcagaga tccacagacc ctggaaatcc tggatattac accctgctcc
     4561 ttcggaggag tttctgtgat cacccccggg accaatacca gcaaccaggt ggctgtgctg
     4621 taccaaggtg ttaactgcac cgaggttcct gtggccatcc acgccgatca gctgacacct
     4681 acttggagag tgtactccac tggctccaat gtgttccaga ccagggccgg atgtctgatc
     4741 ggcgccgagc acgtgaataa cagttacgag tgcgacatcc ctatcggcgc cggcatctgt
     4801 gccagctacc agacccagac aaacagccct gggtctgctt cctctgtagc tagccagagc
     4861 atcatcgcct acaccatgag cctgggcgtg gagaacagcg tggcctattc caacaactct
     4921 atcgccattc ccaccaactt tacaattagc gtcacaacag agatcctgcc cgtgagcatg
     4981 accaagacca gcgtggactg tacaatgtac atctgtggcg acagcactga atgcagcaac
     5041 ctgctgctgc aatacggctc cttttgcacc caactgaacc gggcgctgac cggaatcgcc
     5101 gtggaacagg acaaaaatac ccaggaggtg ttcgcccaag tgaagcagat ctacaagacc
     5161 ccacctatca aggacttcgg cggctttaac tttagccaga ttctccctga tccttctaag
     5221 cctagcaagc ggagctttat cgaggatctg ctgttcaaca aggtcaccct ggccgatgcc
     5281 ggctttatca aacagtatgg cgattgcctg ggcgacatag ccgccagaga tctgatctgc
     5341 gcccagaaat tcaacggcct gacagttctc ccacctctgc tgaccgacga gatgatcgct
     5401 cagtacacct ctgccctgct ggctggcacc atcacatctg ggtggacatt tggcgccggc
     5461 gccgccctgc agatcccctt tgccatgcag atggcctata gattcaacgg aatcggcgtg
     5521 acccagaacg tgctgtatga aaaccagaag ctgatcgcta accagttcaa ttctgccatc
     5581 ggcaagatcc aggactccct ctcctctacc gccagcgccc tgggcaaact gcaggacgtg
     5641 gtgaatcaga acgcccaagc cctgaacacc ctggtgaagc agctcagcag caattttggc
     5701 gccatcagct ctgtgctgaa cgatatcctg tctagactgg accctccaga agccgaagtc
     5761 cagatcgata gactgatcac aggcagactg cagtccctgc aaacctacgt gacccaacag
     5821 ctgatcaggg ccgctgaaat aagagccagc gccaatctcg ccgctaccaa gatgtccgag
     5881 tgtgtgctgg gacagtctaa acgcgttgac ttctgcggca aaggctatca cctgatgagc
     5941 ttcccccaga gcgcgccgca cggcgtggtg ttcctgcatg tgacatacgt gcctgcccaa
     6001 gagaagaatt tcacaaccgc ccctgccatc tgccacgacg gcaaggccca cttcccaaga
     6061 gagggcgttt tcgtttccaa tggcacacac tggttcgtga cacaaagaaa cttctacgaa
     6121 ccccagatta tcaccaccga caacaccttc gtgagtggca attgtgacgt ggtcatcgga
     6181 atcgtgaaca acacagtgta cgaccctctg caacctgagc tggactcttt taaggaagag
     6241 ctggacaagt actttaaaaa ccacaccagc cccgatgtgg acctgggcga catcagtggc
     6301 attaacgcca gcgtggtgaa catccaaaag gaaatcgaca gactgaacga ggtggccaag
     6361 aacctgaacg agtccctgat cgacctgcag gagctcggca aatacgagca gggatctgga
     6421 tacatccccg aggcccccag agatggccag gcctacgtgc ggaaggacgg cgagtgggta
     6481 ctgctgagca cattcctggg cagatccgga ggaggcctgg tgccacagca gagcggaggc
     6541 ctgaacgaca tcttcgaggc ccagaagatc gagtggcacg agggctgatg aggatccaga
     6601 tctgctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     6661 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     6721 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     6781 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg tacccaggtg
     6841 ctgaagaatt gacccggttc ctcctgggcc agaaagaagc aggcacatcc ccttctctgt
     6901 gacacaccct gtccacgccc ctggttctta gttccagccc cactcatagg acactcatag
     6961 ctcaggaggg ctccgccttc aatcccaccc gctaaagtac ttggagcggt ctctccctcc
     7021 ctcatcagcc caccaaacca aacctagcct ccaagagtgg gaagaaatta aagcaagata
     7081 ggctattaag tgcagaggga gagaaaatgc ctccaacatg tgaggaagta atgagagaaa
     7141 tcatagaatt ttaaggccat gatttaaggc catcatggcc ttaatcttcc gcttcctcgc
     7201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8161 cgatctgtct atttcgttca tccatagttg cctgactcgg gggggggggg cgctgaggtc
     8221 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     8281 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
     8341 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     8401 tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat
     8461 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     8521 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     8581 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     8641 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     8701 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     8761 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     8821 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     8881 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     8941 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     9001 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     9061 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     9121 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     9181 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     9241 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     9301 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     9361 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     9421 ttcccccccc ccccattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9541 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9601 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacct
//
