LOCUS       sequence-346986-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein containing D614G 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-D614G-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..26
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(64..86)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     186..205
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        356..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        736..939
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     890..910
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     936..960
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2869..2892
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6604..6648
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6671..6895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7262..7279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7433..8021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7513..7532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8563..9378)
                     /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(8563..9378)
                     /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     9286..9305
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9370..9389
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
       61 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
      121 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
      181 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
      241 cagatgcgta aggagaaaat accgcatcag attggctatt ggccattgca tacgttgtat
      301 ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat
      361 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      421 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      481 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      781 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      841 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      901 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
      961 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
     1021 ctccatcggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     1081 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     1141 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     1201 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctagttaa
     1261 cggtggaggg cagtgtagtc tgagcagtac tcgttgctgc cgcgcgcgcc accagacata
     1321 atagctgaca gactaacaga ctgttccttt ccatgggtct tttctgcagt caccgtcgtc
     1381 gacacgtgtg atcagatatc gcggccgctc tagaccacca tgggatggtc atgtatcatc
     1441 ctttttctag tagcaactgc aaccggtgta cattccgcac cagagctgct gggaggacct
     1501 agcgtgttcc tgtttccccc taagcctaag gacaccctga tgatcagccg gaccccagag
     1561 gtgacatgcg tggtggtgga cgtgtcccac gaggaccccg aggtgaagtt caactggtac
     1621 gtggatggcg tggaggtgca caatgccaag accaagccaa gggaggagca gtacaactct
     1681 acctatcgcg tggtgagcgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag
     1741 tacaagtgta aggtgtctaa taaggccctg cctgccccaa tcgagaagac catcagcaag
     1801 gcaaagggac agccaaggga gcctcaggtg tataccctgc cacccagccg cgacgagctg
     1861 acaaagaacc aggtgtccct gtactgcctg gtgaagggct tctatccttc cgatatcgcc
     1921 gtggagtggg agtctaatgg ccagccagag aacaattaca agaccacacc tccagtgctg
     1981 gactccgatg gctctttctt tctgtattcc aagctgaccg tggataagag ccggtggcag
     2041 cagggcaacg tgtttagctg ttccgtgatg cacgaggccc tgcacaatca ctacacacag
     2101 aagtctctga gcctgtcccc cggcaaggga ggaggaggca gcggcggagg aggctccgga
     2161 ggcggcggct ctggcggcgg cggcagcgcc cctgaactgc tgggaggacc aagcgtgttc
     2221 ctgtttccac ctaagcccaa agataccctg atgatctcca ggacccctga agtcacctgc
     2281 gtggtcgtgg acgtgtctca tgaggacccc gaagtcaagt tcaactggta cgtggacgga
     2341 gtcgaagtcc ataacgccaa gaccaagcct agagaagagc agtacaactc cacctataga
     2401 gtggtgtctg tgctgacagt gctgcatcag gattggctga atggcaagga atataagtgt
     2461 aaggtgagca ataaggccct gcccgcccct atcgagaaga ccatctctaa agccaaagga
     2521 cagccaaggg agccacaggt gtatacactg ccacccagcc gggacgagct gaccaagaac
     2581 caggtgagcc tgacatgtct ggtcaagggc ttctacccca gtgacattgc tgtggagtgg
     2641 gagtccaatg gccagcctga gaacaattat aaaactaccc caccagtgct ggacagcgat
     2701 ggctccttct ttctgaccag caagctgaca gtggataagt cccgctggca gcagggaaac
     2761 gtgttttctt gtagcgtgat gcatgaggcc ctgcataacc actacaccca gaagtccctg
     2821 tctctgagcc caggcaaggg aggcagcgga ggcggaggca gcggaggcct ggaggtgctg
     2881 tttcagggcc ctcagtgcgt gaacctgacc acccgaaccc agctcccacc agcctacacc
     2941 aacagcttta cacggggcgt gtactaccct gacaaggtgt tcagatctag cgtcctgcac
     3001 agcactcagg acctcttcct gccgttcttc agcaacgtga catggttcca cgccatccac
     3061 gtgagcggca caaacggaac caagcggttt gataaccccg tcctgccatt caatgatgga
     3121 gtttacttcg ccagtaccga gaagagtaac atcatccggg gctggatctt cggcaccacc
     3181 ctggatagca aaacacagag cctcctgatc gtgaacaatg ccacgaacgt cgtgatcaag
     3241 gtgtgcgagt tccagttttg caatgatcct ttcctgggtg tgtactacca caagaacaac
     3301 aagagctgga tggaaagcga gttcagagtc tacagcagcg ccaacaactg cacattcgag
     3361 tacgtctctc agccttttct gatggacctt gaggggaaac aaggcaactt caagaacctg
     3421 agagaattcg tgttcaagaa catcgacggc tacttcaaaa tctactccaa gcacacaccc
     3481 atcaacctgg tccgggacct ccctcagggc ttcagcgccc tggaacccct ggtcgacctg
     3541 cccataggca tcaacataac gcggttccaa accctgctgg ccctgcatag atcctacctg
     3601 actcctggcg acagcagcag cggatggacc gccggagctg cagcctacta tgtgggctac
     3661 ctgcaaccta gaaccttcct gctgaagtac aacgagaacg gcacaatcac agacgccgtc
     3721 gactgcgccc tggaccctct ctctgagaca aagtgcaccc tgaagtcctt caccgtggaa
     3781 aagggcatct accagaccag caacttccgg gtgcagccta cagagagcat cgtgcgattt
     3841 ccaaacatta ccaacctctg ccccttcggc gaggtgttta acgccacaag atttgcctcc
     3901 gtttacgcct ggaatagaaa gagaatcagc aattgtgtgg ccgactactc cgtgctgtat
     3961 aacagcgcct ctttcagcac cttcaagtgc tacggcgttt ccccaacaaa gctgaatgac
     4021 ctgtgcttca ccaacgtgta cgccgactcc ttcgtaatta gaggcgatga ggtgcggcag
     4081 atcgcaccag gccagaccgg taagatcgct gactacaact ataagctgcc tgatgatttt
     4141 acaggctgcg tgatcgcctg gaactctaac aacctggata gcaaggtggg cggcaactac
     4201 aactacctgt accggctgtt tcgcaagtct aacctgaaac ctttcgagag agacatctcc
     4261 acagagatct accaggccgg ttctacacct tgtaacgggg tggaaggctt caactgttac
     4321 ttccctctgc aaagctacgg cttccagcct accaatggag tcggctacca gccataccgg
     4381 gtggtcgtgc tgtccttcga gttactccac gcccccgcca ccgtctgcgg tcctaagaag
     4441 tccaccaatc tggttaagaa caaatgcgtg aacttcaact tcaacggcct gaccgggacc
     4501 ggcgtgctga ccgaaagcaa caaaaagttc ctccccttcc agcagttcgg ccgtgatatc
     4561 gctgacacca cagatgccgt cagagatcca cagaccctgg aaatcctgga tattacaccc
     4621 tgctccttcg gaggagtttc tgtgatcacc cccgggacca ataccagcaa ccaggtggct
     4681 gtgctgtacc aaggtgttaa ctgcaccgag gttcctgtgg ccatccacgc cgatcagctg
     4741 acacctactt ggagagtgta ctccactggc tccaatgtgt tccagaccag ggccggatgt
     4801 ctgatcggcg ccgagcacgt gaataacagt tacgagtgcg acatccctat cggcgccggc
     4861 atctgtgcca gctaccagac ccagacaaac agccctgggt ctgcttcctc tgtagctagc
     4921 cagagcatca tcgcctacac catgagcctg ggcgcagaga acagcgtggc ctattccaac
     4981 aactctatcg ccattcccac caactttaca attagcgtca caacagagat cctgcccgtg
     5041 agcatgacca agaccagcgt ggactgtaca atgtacatct gtggcgacag cactgaatgc
     5101 agcaacctgc tgctgcaata cggctccttt tgcacccaac tgaaccgggc gctgaccgga
     5161 atcgccgtgg aacaggacaa aaatacccag gaggtgttcg cccaagtgaa gcagatctac
     5221 aagaccccac ctatcaagga cttcggcggc tttaacttta gccagattct ccctgatcct
     5281 tctaagccta gcaagcggag ctttatcgag gatctgctgt tcaacaaggt caccctggcc
     5341 gatgccggct ttatcaaaca gtatggcgat tgcctgggcg acatagccgc cagagatctg
     5401 atctgcgccc agaaattcaa cggcctgaca gttctcccac ctctgctgac cgacgagatg
     5461 atcgctcagt acacctctgc cctgctggct ggcaccatca catctgggtg gacatttggc
     5521 gccggcgccg ccctgcagat cccctttgcc atgcagatgg cctatagatt caacggaatc
     5581 ggcgtgaccc agaacgtgct gtatgaaaac cagaagctga tcgctaacca gttcaattct
     5641 gccatcggca agatccagga ctccctctcc tctaccgcca gcgccctggg caaactgcag
     5701 gacgtggtga atcagaacgc ccaagccctg aacaccctgg tgaagcagct cagcagcaat
     5761 tttggcgcca tcagctctgt gctgaacgat atcctgtcta gactggaccc tccagaagcc
     5821 gaagtccaga tcgatagact gatcacaggc agactgcagt ccctgcaaac ctacgtgacc
     5881 caacagctga tcagggccgc tgaaataaga gccagcgcca atctcgccgc taccaagatg
     5941 tccgagtgtg tgctgggaca gtctaaacgc gttgacttct gcggcaaagg ctatcacctg
     6001 atgagcttcc cccagagcgc gccgcacggc gtggtgttcc tgcatgtgac atacgtgcct
     6061 gcccaagaga agaatttcac aaccgcccct gccatctgcc acgacggcaa ggcccacttc
     6121 ccaagagagg gcgttttcgt ttccaatggc acacactggt tcgtgacaca aagaaacttc
     6181 tacgaacccc agattatcac caccgacaac accttcgtga gtggcaattg tgacgtggtc
     6241 atcggaatcg tgaacaacac agtgtacgac cctctgcaac ctgagctgga ctcttttaag
     6301 gaagagctgg acaagtactt taaaaaccac accagccccg atgtggacct gggcgacatc
     6361 agtggcatta acgccagcgt ggtgaacatc caaaaggaaa tcgacagact gaacgaggtg
     6421 gccaagaacc tgaacgagtc cctgatcgac ctgcaggagc tcggcaaata cgagcaggga
     6481 tctggataca tccccgaggc ccccagagat ggccaggcct acgtgcggaa ggacggcgag
     6541 tgggtactgc tgagcacatt cctgggcaga tccggaggag gcctggtgcc acagcagagc
     6601 ggaggcctga acgacatctt cgaggcccag aagatcgagt ggcacgaggg ctgatgagga
     6661 tccagatctg ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6721 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6781 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6841 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgggtacc
     6901 caggtgctga agaattgacc cggttcctcc tgggccagaa agaagcaggc acatcccctt
     6961 ctctgtgaca caccctgtcc acgcccctgg ttcttagttc cagccccact cataggacac
     7021 tcatagctca ggagggctcc gccttcaatc ccacccgcta aagtacttgg agcggtctct
     7081 ccctccctca tcagcccacc aaaccaaacc tagcctccaa gagtgggaag aaattaaagc
     7141 aagataggct attaagtgca gagggagaga aaatgcctcc aacatgtgag gaagtaatga
     7201 gagaaatcat agaattttaa ggccatgatt taaggccatc atggccttaa tcttccgctt
     7261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7621 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7861 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actcgggggg ggggggcgct
     8281 gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat
     8341 ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg
     8401 tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct
     8461 gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag
     8521 cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     8581 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     8641 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     8701 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     8761 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     8821 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     8881 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     8941 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     9001 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     9061 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     9121 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     9181 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     9241 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     9301 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc aagacgtttc
     9361 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     9421 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     9481 gtggctttcc cccccccccc attattgaag catttatcag ggttattgtc tcatgagcgg
     9541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9601 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaac
//
