LOCUS       sequence-354493-        9621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the C.37 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 9621)
  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 9621)
  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-C.37-AVI
FEATURES             Location/Qualifiers
     source          1..9621
                     /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             2828..2851
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6542..6586
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6609..6833
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7200..7217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7371..7959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7451..7470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8501..9316)
                     /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(8501..9316)
                     /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     9224..9243
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9308..9327
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9594..9612
                     /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 cgcccaccat
     1381 gggatggtca tgtatcatcc tttttctagt agcaactgca accggcgtac attccgcacc
     1441 agagctgctg ggaggaccta gcgtgttcct gtttccccct aagcctaagg acaccctgat
     1501 gatcagccgg accccagagg tgacatgcgt ggtggtggac gtgtcccacg aggaccccga
     1561 ggtgaagttc aactggtacg tggatggcgt ggaggtgcac aatgccaaga ccaagccaag
     1621 ggaggagcag tacaactcta cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga
     1681 ctggctgaac ggcaaggagt acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat
     1741 cgagaagacc atcagcaagg caaagggaca gccaagggag cctcaggtgt ataccctgcc
     1801 acccagccgc gacgagctga caaagaacca ggtgtccctg tactgcctgg tgaagggctt
     1861 ctatccttcc gatatcgccg tggagtggga gtctaatggc cagccagaga acaattacaa
     1921 gaccacacct ccagtgctgg actccgatgg ctctttcttt ctgtattcca agctgaccgt
     1981 ggataagagc cggtggcagc agggcaacgt gtttagctgt tccgtgatgc acgaggccct
     2041 gcacaatcac tacacacaga agtctctgag cctgtccccc ggcaagggag gaggaggcag
     2101 cggcggagga ggctccggag gcggcggctc tggcggcggc ggcagcgccc ctgaactgct
     2161 gggaggacca agcgtgttcc tgtttccacc taagcccaaa gataccctga tgatctccag
     2221 gacccctgaa gtcacctgcg tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt
     2281 caactggtac gtggacggag tcgaagtcca taacgccaag accaagccta gagaagagca
     2341 gtacaactcc acctatagag tggtgtctgt gctgacagtg ctgcatcagg attggctgaa
     2401 tggcaaggaa tataagtgta aggtgagcaa taaggccctg cccgccccta tcgagaagac
     2461 catctctaaa gccaaaggac agccaaggga gccacaggtg tatacactgc cacccagccg
     2521 ggacgagctg accaagaacc aggtgagcct gacatgtctg gtcaagggct tctaccccag
     2581 tgacattgct gtggagtggg agtccaatgg ccagcctgag aacaattata aaactacccc
     2641 accagtgctg gacagcgatg gctccttctt tctgaccagc aagctgacag tggataagtc
     2701 ccgctggcag cagggaaacg tgttttcttg tagcgtgatg catgaggccc tgcataacca
     2761 ctacacccag aagtccctgt ctctgagccc aggcaaggga ggcagcggag gcggaggcag
     2821 cggaggcctg gaggtgctgt ttcagggccc tcagtgcgtg aacctgacca cccgaaccca
     2881 gctcccacca gcctacacca acagctttac acggggcgtg tactaccctg acaaggtgtt
     2941 cagatctagc gtcctgcaca gcactcagga cctcttcctg ccgttcttca gcaacgtgac
     3001 atggttccac gccatccacg tgagcggcac aaacgtgatc aagcggtttg ataaccccgt
     3061 cctgccattc aatgatggag tttacttcgc cagtaccgag aagagtaaca tcatccgggg
     3121 ctggatcttc ggcaccaccc tggatagcaa aacacagagc ctcctgatcg tgaacaatgc
     3181 cacgaacgtc gtgatcaagg tgtgcgagtt ccagttttgc aatgatcctt tcctgggtgt
     3241 gtactaccac aagaacaaca agagctggat ggaaagcgag ttcagagtct acagcagcgc
     3301 caacaactgc acattcgagt acgtctctca gccttttctg atggaccttg aggggaaaca
     3361 aggcaacttc aagaacctga gagaattcgt gttcaagaac atcgacggct acttcaaaat
     3421 ctactccaag cacacaccca tcaacctggt ccgggacctc cctcagggct tcagcgccct
     3481 ggaacccctg gtcgacctgc ccataggcat caacataacg cggttccaaa ccctgctggc
     3541 cctgcataac agcagcagcg gatggaccgc cggagctgca gcctactatg tgggctacct
     3601 gcaacctaga accttcctgc tgaagtacaa cgagaacggc acaatcacag acgccgtcga
     3661 ctgcgccctg gaccctctct ctgagacaaa gtgcaccctg aagtccttca ccgtggaaaa
     3721 gggcatctac cagaccagca acttccgggt gcagcctaca gagagcatcg tgcgatttcc
     3781 aaacattacc aacctctgcc ccttcggcga ggtgtttaac gccacaagat ttgcctccgt
     3841 ttacgcctgg aatagaaaga gaatcagcaa ttgtgtggcc gactactccg tgctgtataa
     3901 cagcgcctct ttcagcacct tcaagtgcta cggcgtttcc ccaacaaagc tgaatgacct
     3961 gtgcttcacc aacgtgtacg ccgactcctt cgtaattaga ggcgatgagg tgcggcagat
     4021 cgcaccaggc cagaccggta agatcgctga ctacaactat aagctgcctg atgattttac
     4081 aggctgcgtg atcgcctgga actctaacaa cctggatagc aaggtgggcg gcaactacaa
     4141 ctaccagtac cggctgtttc gcaagtctaa cctgaaacct ttcgagagag acatctccac
     4201 agagatctac caggccggtt ctacaccttg taacggggtg gaaggcttca actgttactc
     4261 ccctctgcaa agctacggct tccagcctac caatggagtc ggctaccagc cataccgggt
     4321 ggtcgtgctg tccttcgagt tactccacgc ccccgccacc gtctgcggtc ctaagaagtc
     4381 caccaatctg gttaagaaca aatgcgtgaa cttcaacttc aacggcctga ccgggaccgg
     4441 cgtgctgacc gaaagcaaca aaaagttcct ccccttccag cagttcggcc gtgatatcgc
     4501 tgacaccaca gatgccgtca gagatccaca gaccctggaa atcctggata ttacaccctg
     4561 ctccttcgga ggagtttctg tgatcacccc cgggaccaat accagcaacc aggtggctgt
     4621 gctgtaccaa ggtgttaact gcaccgaggt tcctgtggcc atccacgccg atcagctgac
     4681 acctacttgg agagtgtact ccactggctc caatgtgttc cagaccaggg ccggatgtct
     4741 gatcggcgcc gagcacgtga ataacagtta cgagtgcgac atccctatcg gcgccggcat
     4801 ctgtgccagc taccagaccc agacaaacag ccctgggtct gcttcctctg tagctagcca
     4861 gagcatcatc gcctacacca tgagcctggg cgcagagaac agcgtggcct attccaacaa
     4921 ctctatcgcc attcccacca actttacaat tagcgtcaca acagagatcc tgcccgtgag
     4981 catgaccaag accagcgtgg actgtacaat gtacatctgt ggcgacagca ctgaatgcag
     5041 caacctgctg ctgcaatacg gctccttttg cacccaactg aaccgggcgc tgaccggaat
     5101 cgccgtggaa caggacaaaa atacccagga ggtgttcgcc caagtgaagc agatctacaa
     5161 gaccccacct atcaaggact tcggcggctt taactttagc cagattctcc ctgatccttc
     5221 taagcctagc aagcggagct ttatcgagga tctgctgttc aacaaggtca ccctggccga
     5281 tgccggcttt atcaaacagt atggcgattg cctgggcgac atagccgcca gagatctgat
     5341 ctgcgcccag aaattcaacg gcctgaacgt tctcccacct ctgctgaccg acgagatgat
     5401 cgctcagtac acctctgccc tgctggctgg caccatcaca tctgggtgga catttggcgc
     5461 cggcgccgcc ctgcagatcc cctttgccat gcagatggcc tatagattca acggaatcgg
     5521 cgtgacccag aacgtgctgt atgaaaacca gaagctgatc gctaaccagt tcaattctgc
     5581 catcggcaag atccaggact ccctctcctc taccgccagc gccctgggca aactgcagga
     5641 cgtggtgaat cagaacgccc aagccctgaa caccctggtg aagcagctca gcagcaattt
     5701 tggcgccatc agctctgtgc tgaacgatat cctgtctaga ctggaccctc cagaagccga
     5761 agtccagatc gatagactga tcacaggcag actgcagtcc ctgcaaacct acgtgaccca
     5821 acagctgatc agggccgctg aaataagagc cagcgccaat ctcgccgcta ccaagatgtc
     5881 cgagtgtgtg ctgggacagt ctaaacgcgt tgacttctgc ggcaaaggct atcacctgat
     5941 gagcttcccc cagagcgcgc cgcacggcgt ggtgttcctg catgtgacat acgtgcctgc
     6001 ccaagagaag aatttcacaa ccgcccctgc catctgccac gacggcaagg cccacttccc
     6061 aagagagggc gttttcgttt ccaatggcac acactggttc gtgacacaaa gaaacttcta
     6121 cgaaccccag attatcacca ccgacaacac cttcgtgagt ggcaattgtg acgtggtcat
     6181 cggaatcgtg aacaacacag tgtacgaccc tctgcaacct gagctggact cttttaagga
     6241 agagctggac aagtacttta aaaaccacac cagccccgat gtggacctgg gcgacatcag
     6301 tggcattaac gccagcgtgg tgaacatcca aaaggaaatc gacagactga acgaggtggc
     6361 caagaacctg aacgagtccc tgatcgacct gcaggagctc ggcaaatacg agcagggatc
     6421 tggatacatc cccgaggccc ccagagatgg ccaggcctac gtgcggaagg acggcgagtg
     6481 ggtactgctg agcacattcc tgggcagatc cggaggaggc ctggtgccac agcagagcgg
     6541 aggcctgaac gacatcttcg aggcccagaa gatcgagtgg cacgagggct gatgaggatc
     6601 cagatctgct gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6661 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6721 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     6781 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tgggtaccca
     6841 ggtgctgaag aattgacccg gttcctcctg ggccagaaag aagcaggcac atccccttct
     6901 ctgtgacaca ccctgtccac gcccctggtt cttagttcca gccccactca taggacactc
     6961 atagctcagg agggctccgc cttcaatccc acccgctaaa gtacttggag cggtctctcc
     7021 ctccctcatc agcccaccaa accaaaccta gcctccaaga gtgggaagaa attaaagcaa
     7081 gataggctat taagtgcaga gggagagaaa atgcctccaa catgtgagga agtaatgaga
     7141 gaaatcatag aattttaagg ccatgattta aggccatcat ggccttaatc ttccgcttcc
     7201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7801 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tcgggggggg ggggcgctga
     8221 ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc
     8281 agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg
     8341 attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga
     8401 tccttcaact cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg
     8461 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     8521 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     8581 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     8641 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     8701 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     8761 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     8821 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     8881 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     8941 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     9001 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     9061 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     9121 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     9181 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     9241 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
     9301 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     9361 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     9421 ggctttcccc ccccccccat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9541 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9601 gtatcacgag gccctttcgt c
//
