LOCUS       sequence-347063-        9639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.351 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 9639)
  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 9639)
  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.1.351-AVI
FEATURES             Location/Qualifiers
     source          1..9639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..71)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     171..190
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        341..720
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        721..924
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     875..895
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     921..945
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2854..2877
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6580..6624
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6647..6871
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7238..7255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7409..7997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7489..7508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8539..9354)
                     /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(8539..9354)
                     /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     9262..9281
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9346..9365
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     join(9632..9639,1..11)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
       61 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      121 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      181 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      241 aaaataccgc atcagattgg ctattggcca ttgcatacgt tgtatccata tcataatatg
      301 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
      361 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      421 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      481 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      541 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      601 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      661 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      721 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      781 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      841 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      901 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      961 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctcca tcggctcgca
     1021 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     1081 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     1141 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     1201 ggctctccac gctttgcctg accctgcttg ctcaactcta gttaacggtg gagggcagtg
     1261 tagtctgagc agtactcgtt gctgccgcgc gcgccaccag acataatagc tgacagacta
     1321 acagactgtt cctttccatg ggtcttttct gcagtcaccg tcgtcgacac gtgtgatcag
     1381 atatcgcggc cgctctagac caccatggga tggtcatgta tcatcctttt tctagtagca
     1441 actgcaaccg gtgtacattc cgcaccagag ctgctgggag gacctagcgt gttcctgttt
     1501 ccccctaagc ctaaggacac cctgatgatc agccggaccc cagaggtgac atgcgtggtg
     1561 gtggacgtgt cccacgagga ccccgaggtg aagttcaact ggtacgtgga tggcgtggag
     1621 gtgcacaatg ccaagaccaa gccaagggag gagcagtaca actctaccta tcgcgtggtg
     1681 agcgtgctga cagtgctgca ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg
     1741 tctaataagg ccctgcctgc cccaatcgag aagaccatca gcaaggcaaa gggacagcca
     1801 agggagcctc aggtgtatac cctgccaccc agccgcgacg agctgacaaa gaaccaggtg
     1861 tccctgtact gcctggtgaa gggcttctat ccttccgata tcgccgtgga gtgggagtct
     1921 aatggccagc cagagaacaa ttacaagacc acacctccag tgctggactc cgatggctct
     1981 ttctttctgt attccaagct gaccgtggat aagagccggt ggcagcaggg caacgtgttt
     2041 agctgttccg tgatgcacga ggccctgcac aatcactaca cacagaagtc tctgagcctg
     2101 tcccccggca agggaggagg aggcagcggc ggaggaggct ccggaggcgg cggctctggc
     2161 ggcggcggca gcgcccctga actgctggga ggaccaagcg tgttcctgtt tccacctaag
     2221 cccaaagata ccctgatgat ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg
     2281 tctcatgagg accccgaagt caagttcaac tggtacgtgg acggagtcga agtccataac
     2341 gccaagacca agcctagaga agagcagtac aactccacct atagagtggt gtctgtgctg
     2401 acagtgctgc atcaggattg gctgaatggc aaggaatata agtgtaaggt gagcaataag
     2461 gccctgcccg cccctatcga gaagaccatc tctaaagcca aaggacagcc aagggagcca
     2521 caggtgtata cactgccacc cagccgggac gagctgacca agaaccaggt gagcctgaca
     2581 tgtctggtca agggcttcta ccccagtgac attgctgtgg agtgggagtc caatggccag
     2641 cctgagaaca attataaaac taccccacca gtgctggaca gcgatggctc cttctttctg
     2701 accagcaagc tgacagtgga taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc
     2761 gtgatgcatg aggccctgca taaccactac acccagaagt ccctgtctct gagcccaggc
     2821 aagggaggca gcggaggcgg aggcagcgga ggcctggagg tgctgtttca gggccctcag
     2881 tgcgtgaact tcaccacccg aacccagctc ccaccagcct acaccaacag ctttacacgg
     2941 ggcgtgtact accctgacaa ggtgttcaga tctagcgtcc tgcacagcac tcaggacctc
     3001 ttcctgccgt tcttcagcaa cgtgacatgg ttccacgcca tccacgtgag cggcacaaac
     3061 ggaaccaagc ggtttgctaa ccccgtcctg ccattcaatg atggagttta cttcgccagt
     3121 accgagaaga gtaacatcat ccggggctgg atcttcggca ccaccctgga tagcaaaaca
     3181 cagagcctcc tgatcgtgaa caatgccacg aacgtcgtga tcaaggtgtg cgagttccag
     3241 ttttgcaatg atcctttcct gggtgtgtac taccacaaga acaacaagag ctggatggaa
     3301 agcgagttca gagtctacag cagcgccaac aactgcacat tcgagtacgt ctctcagcct
     3361 tttctgatgg accttgaggg gaaacaaggc aacttcaaga acctgagaga attcgtgttc
     3421 aagaacatcg acggctactt caaaatctac tccaagcaca cacccatcaa cctggtccgg
     3481 ggcctccctc agggcttcag cgccctggaa cccctggtcg acctgcccat aggcatcaac
     3541 ataacgcggt tccaaaccct gcatatctcc tacctgactc ctggcgacag cagcagcgga
     3601 tggaccgccg gagctgcagc ctactatgtg ggctacctgc aacctagaac cttcctgctg
     3661 aagtacaacg agaacggcac aatcacagac gccgtcgact gcgccctgga ccctctctct
     3721 gagacaaagt gcaccctgaa gtccttcacc gtggaaaagg gcatctacca gaccagcaac
     3781 ttccgggtgc agcctacaga gagcatcgtg cgatttccaa acattaccaa cctctgcccc
     3841 ttcggcgagg tgtttaacgc cacaagattt gcctccgttt acgcctggaa tagaaagaga
     3901 atcagcaatt gtgtggccga ctactccgtg ctgtataaca gcgcctcttt cagcaccttc
     3961 aagtgctacg gcgtttcccc aacaaagctg aatgacctgt gcttcaccaa cgtgtacgcc
     4021 gactccttcg taattagagg cgatgaggtg cggcagatcg caccaggcca gaccggtaac
     4081 atcgctgact acaactataa gctgcctgat gattttacag gctgcgtgat cgcctggaac
     4141 tctaacaacc tggatagcaa ggtgggcggc aactacaact acctgtaccg gctgtttcgc
     4201 aagtctaacc tgaaaccttt cgagagagac atctccacag agatctacca ggccggttct
     4261 acaccttgta acggggtgaa aggcttcaac tgttacttcc ctctgcaaag ctacggcttc
     4321 cagcctacct acggagtcgg ctaccagcca taccgggtgg tcgtgctgtc cttcgagtta
     4381 ctccacgccc ccgccaccgt ctgcggtcct aagaagtcca ccaatctggt taagaacaaa
     4441 tgcgtgaact tcaacttcaa cggcctgacc gggaccggcg tgctgaccga aagcaacaaa
     4501 aagttcctcc ccttccagca gttcggccgt gatatcgctg acaccacaga tgccgtcaga
     4561 gatccacaga ccctggaaat cctggatatt acaccctgct ccttcggagg agtttctgtg
     4621 atcacccccg ggaccaatac cagcaaccag gtggctgtgc tgtaccaagg tgttaactgc
     4681 accgaggttc ctgtggccat ccacgccgat cagctgacac ctacttggag agtgtactcc
     4741 actggctcca atgtgttcca gaccagggcc ggatgtctga tcggcgccga gcacgtgaat
     4801 aacagttacg agtgcgacat ccctatcggc gccggcatct gtgccagcta ccagacccag
     4861 acaaacagcc ctgggtctgc ttcctctgta gctagccaga gcatcatcgc ctacaccatg
     4921 agcctgggcg tggagaacag cgtggcctat tccaacaact ctatcgccat tcccaccaac
     4981 tttacaatta gcgtcacaac agagatcctg cccgtgagca tgaccaagac cagcgtggac
     5041 tgtacaatgt acatctgtgg cgacagcact gaatgcagca acctgctgct gcaatacggc
     5101 tccttttgca cccaactgaa ccgggcgctg accggaatcg ccgtggaaca ggacaaaaat
     5161 acccaggagg tgttcgccca agtgaagcag atctacaaga ccccacctat caaggacttc
     5221 ggcggcttta actttagcca gattctccct gatccttcta agcctagcaa gcggagcttt
     5281 atcgaggatc tgctgttcaa caaggtcacc ctggccgatg ccggctttat caaacagtat
     5341 ggcgattgcc tgggcgacat agccgccaga gatctgatct gcgcccagaa attcaacggc
     5401 ctgacagttc tcccacctct gctgaccgac gagatgatcg ctcagtacac ctctgccctg
     5461 ctggctggca ccatcacatc tgggtggaca tttggcgccg gcgccgccct gcagatcccc
     5521 tttgccatgc agatggccta tagattcaac ggaatcggcg tgacccagaa cgtgctgtat
     5581 gaaaaccaga agctgatcgc taaccagttc aattctgcca tcggcaagat ccaggactcc
     5641 ctctcctcta ccgccagcgc cctgggcaaa ctgcaggacg tggtgaatca gaacgcccaa
     5701 gccctgaaca ccctggtgaa gcagctcagc agcaattttg gcgccatcag ctctgtgctg
     5761 aacgatatcc tgtctagact ggaccctcca gaagccgaag tccagatcga tagactgatc
     5821 acaggcagac tgcagtccct gcaaacctac gtgacccaac agctgatcag ggccgctgaa
     5881 ataagagcca gcgccaatct cgccgctacc aagatgtccg agtgtgtgct gggacagtct
     5941 aaacgcgttg acttctgcgg caaaggctat cacctgatga gcttccccca gagcgcgccg
     6001 cacggcgtgg tgttcctgca tgtgacatac gtgcctgccc aagagaagaa tttcacaacc
     6061 gcccctgcca tctgccacga cggcaaggcc cacttcccaa gagagggcgt tttcgtttcc
     6121 aatggcacac actggttcgt gacacaaaga aacttctacg aaccccagat tatcaccacc
     6181 gacaacacct tcgtgagtgg caattgtgac gtggtcatcg gaatcgtgaa caacacagtg
     6241 tacgaccctc tgcaacctga gctggactct tttaaggaag agctggacaa gtactttaaa
     6301 aaccacacca gccccgatgt ggacctgggc gacatcagtg gcattaacgc cagcgtggtg
     6361 aacatccaaa aggaaatcga cagactgaac gaggtggcca agaacctgaa cgagtccctg
     6421 atcgacctgc aggagctcgg caaatacgag cagggatctg gatacatccc cgaggccccc
     6481 agagatggcc aggcctacgt gcggaaggac ggcgagtggg tactgctgag cacattcctg
     6541 ggcagatccg gaggaggcct ggtgccacag cagagcggag gcctgaacga catcttcgag
     6601 gcccagaaga tcgagtggca cgagggctga tgaggatcca gatctgctgt gccttctagt
     6661 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6721 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6781 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6841 aggcatgctg gggatgcggt gggctctatg ggtacccagg tgctgaagaa ttgacccggt
     6901 tcctcctggg ccagaaagaa gcaggcacat ccccttctct gtgacacacc ctgtccacgc
     6961 ccctggttct tagttccagc cccactcata ggacactcat agctcaggag ggctccgcct
     7021 tcaatcccac ccgctaaagt acttggagcg gtctctccct ccctcatcag cccaccaaac
     7081 caaacctagc ctccaagagt gggaagaaat taaagcaaga taggctatta agtgcagagg
     7141 gagagaaaat gcctccaaca tgtgaggaag taatgagaga aatcatagaa ttttaaggcc
     7201 atgatttaag gccatcatgg ccttaatctt ccgcttcctc gctcactgac tcgctgcgct
     7261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8221 catccatagt tgcctgactc gggggggggg ggcgctgagg tctgcctcgt gaagaaggtg
     8281 ttgctgactc ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac
     8341 ggttgatgag agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca
     8401 cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     8461 gatttattca acaaagccgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
     8521 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     8581 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     8641 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     8701 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     8761 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     8821 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     8881 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     8941 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     9001 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     9061 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     9121 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     9181 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     9241 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     9301 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
     9361 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     9421 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcccccc ccccccatta
     9481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9601 aaccattatt atcatgacat taacctataa aaataggcg
//
