LOCUS       sequence-347140-        9648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.617 variant containing
            E154K-L452R-E484Q-D614G-P681R mutations 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.1.617-E154K-L452R-E484Q-D614G-P681R-AVI
FEATURES             Location/Qualifiers
     source          1..9648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..58)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     158..177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        328..707
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        708..911
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     862..882
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     908..932
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2841..2864
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6576..6620
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6643..6867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7234..7251)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7405..7993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7485..7504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8535..9350)
                     /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(8535..9350)
                     /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     9258..9277
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9342..9361
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9628..9646
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
       61 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      121 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      181 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      241 agattggcta ttggccattg catacgttgt atccatatca taatatgtac atttatattg
      301 gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat taatagtaat
      361 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      421 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      481 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      541 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      601 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      661 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      721 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      781 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      841 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      901 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
      961 acctccatag aagacaccgg gaccgatcca gcctccatcg gctcgcatct ctccttcacg
     1021 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     1081 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     1141 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     1201 ttgcctgacc ctgcttgctc aactctagtt aacggtggag ggcagtgtag tctgagcagt
     1261 actcgttgct gccgcgcgcg ccaccagaca taatagctga cagactaaca gactgttcct
     1321 ttccatgggt cttttctgca gtcaccgtcg tcgacacgtg tgatcagata tcgcggccgc
     1381 tctagaccac catgggatgg tcatgtatca tcctttttct agtagcaact gcaaccggtg
     1441 tacattccgc accagagctg ctgggaggac ctagcgtgtt cctgtttccc cctaagccta
     1501 aggacaccct gatgatcagc cggaccccag aggtgacatg cgtggtggtg gacgtgtccc
     1561 acgaggaccc cgaggtgaag ttcaactggt acgtggatgg cgtggaggtg cacaatgcca
     1621 agaccaagcc aagggaggag cagtacaact ctacctatcg cgtggtgagc gtgctgacag
     1681 tgctgcacca ggactggctg aacggcaagg agtacaagtg taaggtgtct aataaggccc
     1741 tgcctgcccc aatcgagaag accatcagca aggcaaaggg acagccaagg gagcctcagg
     1801 tgtataccct gccacccagc cgcgacgagc tgacaaagaa ccaggtgtcc ctgtactgcc
     1861 tggtgaaggg cttctatcct tccgatatcg ccgtggagtg ggagtctaat ggccagccag
     1921 agaacaatta caagaccaca cctccagtgc tggactccga tggctctttc tttctgtatt
     1981 ccaagctgac cgtggataag agccggtggc agcagggcaa cgtgtttagc tgttccgtga
     2041 tgcacgaggc cctgcacaat cactacacac agaagtctct gagcctgtcc cccggcaagg
     2101 gaggaggagg cagcggcgga ggaggctccg gaggcggcgg ctctggcggc ggcggcagcg
     2161 cccctgaact gctgggagga ccaagcgtgt tcctgtttcc acctaagccc aaagataccc
     2221 tgatgatctc caggacccct gaagtcacct gcgtggtcgt ggacgtgtct catgaggacc
     2281 ccgaagtcaa gttcaactgg tacgtggacg gagtcgaagt ccataacgcc aagaccaagc
     2341 ctagagaaga gcagtacaac tccacctata gagtggtgtc tgtgctgaca gtgctgcatc
     2401 aggattggct gaatggcaag gaatataagt gtaaggtgag caataaggcc ctgcccgccc
     2461 ctatcgagaa gaccatctct aaagccaaag gacagccaag ggagccacag gtgtatacac
     2521 tgccacccag ccgggacgag ctgaccaaga accaggtgag cctgacatgt ctggtcaagg
     2581 gcttctaccc cagtgacatt gctgtggagt gggagtccaa tggccagcct gagaacaatt
     2641 ataaaactac cccaccagtg ctggacagcg atggctcctt ctttctgacc agcaagctga
     2701 cagtggataa gtcccgctgg cagcagggaa acgtgttttc ttgtagcgtg atgcatgagg
     2761 ccctgcataa ccactacacc cagaagtccc tgtctctgag cccaggcaag ggaggcagcg
     2821 gaggcggagg cagcggaggc ctggaggtgc tgtttcaggg ccctcagtgc gtgaacctga
     2881 ccacccgaac ccagctccca ccagcctaca ccaacagctt tacacggggc gtgtactacc
     2941 ctgacaaggt gttcagatct agcgtcctgc acagcactca ggacctcttc ctgccgttct
     3001 tcagcaacgt gacatggttc cacgccatcc acgtgagcgg cacaaacgga accaagcggt
     3061 ttgataaccc cgtcctgcca ttcaatgatg gagtttactt cgccagtacc gagaagagta
     3121 acatcatccg gggctggatc ttcggcacca ccctggatag caaaacacag agcctcctga
     3181 tcgtgaacaa tgccacgaac gtcgtgatca aggtgtgcga gttccagttt tgcaatgatc
     3241 ctttcctggg tgtgtactac cacaagaaca acaagagctg gatgaagagc gagttcagag
     3301 tctacagcag cgccaacaac tgcacattcg agtacgtctc tcagcctttt ctgatggacc
     3361 ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt cgtgttcaag aacatcgacg
     3421 gctacttcaa aatctactcc aagcacacac ccatcaacct ggtccgggac ctccctcagg
     3481 gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg catcaacata acgcggttcc
     3541 aaaccctgct ggccctgcat agatcctacc tgactcctgg cgacagcagc agcggatgga
     3601 ccgccggagc tgcagcctac tatgtgggct acctgcaacc tagaaccttc ctgctgaagt
     3661 acaacgagaa cggcacaatc acagacgccg tcgactgcgc cctggaccct ctctctgaga
     3721 caaagtgcac cctgaagtcc ttcaccgtgg aaaagggcat ctaccagacc agcaacttcc
     3781 gggtgcagcc tacagagagc atcgtgcgat ttccaaacat taccaacctc tgccccttcg
     3841 gcgaggtgtt taacgccaca agatttgcct ccgtttacgc ctggaataga aagagaatca
     3901 gcaattgtgt ggccgactac tccgtgctgt ataacagcgc ctctttcagc accttcaagt
     3961 gctacggcgt ttccccaaca aagctgaatg acctgtgctt caccaacgtg tacgccgact
     4021 ccttcgtaat tagaggcgat gaggtgcggc agatcgcacc aggccagacc ggtaagatcg
     4081 ctgactacaa ctataagctg cctgatgatt ttacaggctg cgtgatcgcc tggaactcta
     4141 acaacctgga tagcaaggtg ggcggcaact acaactacag ataccggctg tttcgcaagt
     4201 ctaacctgaa acctttcgag agagacatct ccacagagat ctaccaggcc ggttctacac
     4261 cttgtaacgg ggtgcagggc ttcaactgtt acttccctct gcaaagctac ggcttccagc
     4321 ctaccaatgg agtcggctac cagccatacc gggtggtcgt gctgtccttc gagttactcc
     4381 acgcccccgc caccgtctgc ggtcctaaga agtccaccaa tctggttaag aacaaatgcg
     4441 tgaacttcaa cttcaacggc ctgaccggga ccggcgtgct gaccgaaagc aacaaaaagt
     4501 tcctcccctt ccagcagttc ggccgtgata tcgctgacac cacagatgcc gtcagagatc
     4561 cacagaccct ggaaatcctg gatattacac cctgctcctt cggaggagtt tctgtgatca
     4621 cccccgggac caataccagc aaccaggtgg ctgtgctgta ccaaggtgtt aactgcaccg
     4681 aggttcctgt ggccatccac gccgatcagc tgacacctac ttggagagtg tactccactg
     4741 gctccaatgt gttccagacc agggccggat gtctgatcgg cgccgagcac gtgaataaca
     4801 gttacgagtg cgacatccct atcggcgccg gcatctgtgc cagctaccag acccagacaa
     4861 acagcagagg gtctgcttcc tctgtagcta gccagagcat catcgcctac accatgagcc
     4921 tgggcgcaga gaacagcgtg gcctattcca acaactctat cgccattccc accaacttta
     4981 caattagcgt cacaacagag atcctgcccg tgagcatgac caagaccagc gtggactgta
     5041 caatgtacat ctgtggcgac agcactgaat gcagcaacct gctgctgcaa tacggctcct
     5101 tttgcaccca actgaaccgg gcgctgaccg gaatcgccgt ggaacaggac aaaaataccc
     5161 aggaggtgtt cgcccaagtg aagcagatct acaagacccc acctatcaag gacttcggcg
     5221 gctttaactt tagccagatt ctccctgatc cttctaagcc tagcaagcgg agctttatcg
     5281 aggatctgct gttcaacaag gtcaccctgg ccgatgccgg ctttatcaaa cagtatggcg
     5341 attgcctggg cgacatagcc gccagagatc tgatctgcgc ccagaaattc aacggcctga
     5401 cagttctccc acctctgctg accgacgaga tgatcgctca gtacacctct gccctgctgg
     5461 ctggcaccat cacatctggg tggacatttg gcgccggcgc cgccctgcag atcccctttg
     5521 ccatgcagat ggcctataga ttcaacggaa tcggcgtgac ccagaacgtg ctgtatgaaa
     5581 accagaagct gatcgctaac cagttcaatt ctgccatcgg caagatccag gactccctct
     5641 cctctaccgc cagcgccctg ggcaaactgc aggacgtggt gaatcagaac gcccaagccc
     5701 tgaacaccct ggtgaagcag ctcagcagca attttggcgc catcagctct gtgctgaacg
     5761 atatcctgtc tagactggac cctccagaag ccgaagtcca gatcgataga ctgatcacag
     5821 gcagactgca gtccctgcaa acctacgtga cccaacagct gatcagggcc gctgaaataa
     5881 gagccagcgc caatctcgcc gctaccaaga tgtccgagtg tgtgctggga cagtctaaac
     5941 gcgttgactt ctgcggcaaa ggctatcacc tgatgagctt cccccagagc gcgccgcacg
     6001 gcgtggtgtt cctgcatgtg acatacgtgc ctgcccaaga gaagaatttc acaaccgccc
     6061 ctgccatctg ccacgacggc aaggcccact tcccaagaga gggcgttttc gtttccaatg
     6121 gcacacactg gttcgtgaca caaagaaact tctacgaacc ccagattatc accaccgaca
     6181 acaccttcgt gagtggcaat tgtgacgtgg tcatcggaat cgtgaacaac acagtgtacg
     6241 accctctgca acctgagctg gactctttta aggaagagct ggacaagtac tttaaaaacc
     6301 acaccagccc cgatgtggac ctgggcgaca tcagtggcat taacgccagc gtggtgaaca
     6361 tccaaaagga aatcgacaga ctgaacgagg tggccaagaa cctgaacgag tccctgatcg
     6421 acctgcagga gctcggcaaa tacgagcagg gatctggata catccccgag gcccccagag
     6481 atggccaggc ctacgtgcgg aaggacggcg agtgggtact gctgagcaca ttcctgggca
     6541 gatccggagg aggcctggtg ccacagcaga gcggaggcct gaacgacatc ttcgaggccc
     6601 agaagatcga gtggcacgag ggctgatgag gatccagatc tgctgtgcct tctagttgcc
     6661 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     6721 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     6781 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     6841 atgctgggga tgcggtgggc tctatgggta cccaggtgct gaagaattga cccggttcct
     6901 cctgggccag aaagaagcag gcacatcccc ttctctgtga cacaccctgt ccacgcccct
     6961 ggttcttagt tccagcccca ctcataggac actcatagct caggagggct ccgccttcaa
     7021 tcccacccgc taaagtactt ggagcggtct ctccctccct catcagccca ccaaaccaaa
     7081 cctagcctcc aagagtggga agaaattaaa gcaagatagg ctattaagtg cagagggaga
     7141 gaaaatgcct ccaacatgtg aggaagtaat gagagaaatc atagaatttt aaggccatga
     7201 tttaaggcca tcatggcctt aatcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7261 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7321 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7981 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8041 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8101 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8161 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8221 catagttgcc tgactcgggg ggggggggcg ctgaggtctg cctcgtgaag aaggtgttgc
     8281 tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt
     8341 gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga
     8401 acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt
     8461 tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa
     8521 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     8581 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     8641 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     8701 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     8761 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     8821 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     8881 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     8941 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     9001 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     9061 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     9121 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     9181 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     9241 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     9301 gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     9361 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     9421 gcaatgtaac atcagagatt ttgagacaca acgtggcttt cccccccccc ccattattga
     9481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     9601 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccc
//
