LOCUS       sequence-347146-        9642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.617.2 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 9642)
  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 9642)
  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.1.617.2-AVI
FEATURES             Location/Qualifiers
     source          1..9642
                     /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             2834..2857
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6563..6607
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6630..6854
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7221..7238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7392..7980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7472..7491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8522..9337)
                     /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(8522..9337)
                     /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     9245..9264
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9329..9348
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9615..9633
                     /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 cgctctagac
     1381 caccatggga tggtcatgta tcatcctttt tctagtagca actgcaaccg gtgtacattc
     1441 cgcaccagag ctgctgggag gacctagcgt gttcctgttt ccccctaagc ctaaggacac
     1501 cctgatgatc agccggaccc cagaggtgac atgcgtggtg gtggacgtgt cccacgagga
     1561 ccccgaggtg aagttcaact ggtacgtgga tggcgtggag gtgcacaatg ccaagaccaa
     1621 gccaagggag gagcagtaca actctaccta tcgcgtggtg agcgtgctga cagtgctgca
     1681 ccaggactgg ctgaacggca aggagtacaa gtgtaaggtg tctaataagg ccctgcctgc
     1741 cccaatcgag aagaccatca gcaaggcaaa gggacagcca agggagcctc aggtgtatac
     1801 cctgccaccc agccgcgacg agctgacaaa gaaccaggtg tccctgtact gcctggtgaa
     1861 gggcttctat ccttccgata tcgccgtgga gtgggagtct aatggccagc cagagaacaa
     1921 ttacaagacc acacctccag tgctggactc cgatggctct ttctttctgt attccaagct
     1981 gaccgtggat aagagccggt ggcagcaggg caacgtgttt agctgttccg tgatgcacga
     2041 ggccctgcac aatcactaca cacagaagtc tctgagcctg tcccccggca agggaggagg
     2101 aggcagcggc ggaggaggct ccggaggcgg cggctctggc ggcggcggca gcgcccctga
     2161 actgctggga ggaccaagcg tgttcctgtt tccacctaag cccaaagata ccctgatgat
     2221 ctccaggacc cctgaagtca cctgcgtggt cgtggacgtg tctcatgagg accccgaagt
     2281 caagttcaac tggtacgtgg acggagtcga agtccataac gccaagacca agcctagaga
     2341 agagcagtac aactccacct atagagtggt gtctgtgctg acagtgctgc atcaggattg
     2401 gctgaatggc aaggaatata agtgtaaggt gagcaataag gccctgcccg cccctatcga
     2461 gaagaccatc tctaaagcca aaggacagcc aagggagcca caggtgtata cactgccacc
     2521 cagccgggac gagctgacca agaaccaggt gagcctgaca tgtctggtca agggcttcta
     2581 ccccagtgac attgctgtgg agtgggagtc caatggccag cctgagaaca attataaaac
     2641 taccccacca gtgctggaca gcgatggctc cttctttctg accagcaagc tgacagtgga
     2701 taagtcccgc tggcagcagg gaaacgtgtt ttcttgtagc gtgatgcatg aggccctgca
     2761 taaccactac acccagaagt ccctgtctct gagcccaggc aagggaggca gcggaggcgg
     2821 aggcagcgga ggcctggagg tgctgtttca gggccctcag tgcgtgaacc tgagaacccg
     2881 aacccagctc ccaccagcct acaccaacag ctttacacgg ggcgtgtact accctgacaa
     2941 ggtgttcaga tctagcgtcc tgcacagcac tcaggacctc ttcctgccgt tcttcagcaa
     3001 cgtgacatgg ttccacgcca tccacgtgag cggcacaaac ggaaccaagc ggtttgataa
     3061 ccccgtcctg ccattcaatg atggagttta cttcgccagt accgagaaga gtaacatcat
     3121 ccggggctgg atcttcggca ccaccctgga tagcaaaaca cagagcctcc tgatcgtgaa
     3181 caatgccacg aacgtcgtga tcaaggtgtg cgagttccag ttttgcaatg atcctttcct
     3241 ggacgtgtac taccacaaga acaacaagag ctggatggaa agcggcgtct 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 cctgcataga tcctacctga ctcctggcga cagcagcagc ggatggaccg ccggagctgc
     3601 agcctactat gtgggctacc tgcaacctag aaccttcctg ctgaagtaca acgagaacgg
     3661 cacaatcaca gacgccgtcg actgcgccct ggaccctctc tctgagacaa agtgcaccct
     3721 gaagtccttc accgtggaaa agggcatcta ccagaccagc aacttccggg tgcagcctac
     3781 agagagcatc gtgcgatttc caaacattac caacctctgc cccttcggcg aggtgtttaa
     3841 cgccacaaga tttgcctccg tttacgcctg gaatagaaag agaatcagca attgtgtggc
     3901 cgactactcc gtgctgtata acagcgcctc tttcagcacc ttcaagtgct acggcgtttc
     3961 cccaacaaag ctgaatgacc tgtgcttcac caacgtgtac gccgactcct tcgtaattag
     4021 aggcgatgag gtgcggcaga tcgcaccagg ccagaccggt aagatcgctg actacaacta
     4081 taagctgcct gatgatttta caggctgcgt gatcgcctgg aactctaaca acctggatag
     4141 caaggtgggc ggcaactaca actacagata ccggctgttt cgcaagtcta acctgaaacc
     4201 tttcgagaga gacatctcca cagagatcta ccaggccggt tctaagcctt gtaacggggt
     4261 ggaaggcttc aactgttact tccctctgca aagctacggc ttccagccta ccaatggagt
     4321 cggctaccag ccataccggg tggtcgtgct gtccttcgag ttactccacg cccccgccac
     4381 cgtctgcggt cctaagaagt ccaccaatct ggttaagaac aaatgcgtga acttcaactt
     4441 caacggcctg actgggaccg gcgtgctgac cgaaagcaac aaaaagttcc tccccttcca
     4501 gcagttcggc cgtgatatcg ctgacaccac agatgccgtc agagatccac agaccctgga
     4561 aatcctggat attacaccct gctccttcgg aggagtttct gtgatcaccc ccgggaccaa
     4621 taccagcaac caggtggctg tgctgtacca aggtgttaac tgcaccgagg ttcctgtggc
     4681 catccacgcc gatcagctga cacctacttg gagagtgtac tccactggct ccaatgtgtt
     4741 ccagaccagg gccggatgtc tgatcggcgc cgagcacgtg aataacagtt acgagtgcga
     4801 catccctatc ggcgccggca tctgtgccag ctaccagacc cagacaaaca gcagagggtc
     4861 tgcttcctct gtagctagcc agagcatcat cgcctacacc atgagcctgg gcgcagagaa
     4921 cagcgtggcc tattccaaca actctatcgc cattcccacc aactttacaa ttagcgtcac
     4981 aacagagatc ctgcccgtga gcatgaccaa gaccagcgtg gactgtacaa tgtacatctg
     5041 tggcgacagc actgaatgca gcaacctgct gctgcaatac ggctcctttt gcacccaact
     5101 gaaccgggcg ctgaccggaa tcgccgtgga acaggacaaa aatacccagg aggtgttcgc
     5161 ccaagtgaag cagatctaca agaccccacc tatcaaggac ttcggcggct ttaactttag
     5221 ccagattctc cctgatcctt ctaagcctag caagcggagc tttatcgagg atctgctgtt
     5281 caacaaggtc accctggccg atgccggctt tatcaaacag tatggcgatt gcctgggcga
     5341 catagccgcc agagatctga tctgcgccca gaaattcaac ggcctgacag ttctcccacc
     5401 tctgctgacc gacgagatga tcgctcagta cacctctgcc ctgctggctg gcaccatcac
     5461 atctgggtgg acatttggcg ccggcgccgc cctgcagatc ccctttgcca tgcagatggc
     5521 ctatagattc aacggaatcg gcgtgaccca gaacgtgctg tatgaaaacc agaagctgat
     5581 cgctaaccag ttcaattctg ccatcggcaa gatccaggac tccctctcct ctaccgccag
     5641 cgccctgggc aaactgcaga acgtggtgaa tcagaacgcc caagccctga acaccctggt
     5701 gaagcagctc agcagcaatt ttggcgccat cagctctgtg ctgaacgata tcctgtctag
     5761 actggaccct ccagaagccg aagtccagat cgatagactg atcacaggca gactgcagtc
     5821 cctgcaaacc tacgtgaccc aacagctgat cagggccgct gaaataagag ccagcgccaa
     5881 tctcgccgct accaagatgt ccgagtgtgt gctgggacag tctaaacgcg ttgacttctg
     5941 cggcaaaggc tatcacctga tgagcttccc ccagagcgcg ccgcacggcg tggtgttcct
     6001 gcatgtgaca tacgtgcctg cccaagagaa gaatttcaca accgcccctg ccatctgcca
     6061 cgacggcaag gcccacttcc caagagaggg cgttttcgtt tccaatggca cacactggtt
     6121 cgtgacacaa agaaacttct acgaacccca gattatcacc accgacaaca ccttcgtgag
     6181 tggcaattgt gacgtggtca tcggaatcgt gaacaacaca gtgtacgacc ctctgcaacc
     6241 tgagctggac tcttttaagg aagagctgga caagtacttt aaaaaccaca ccagccccga
     6301 tgtggacctg ggcgacatca gtggcattaa cgccagcgtg gtgaacatcc aaaaggaaat
     6361 cgacagactg aacgaggtgg ccaagaacct gaacgagtcc ctgatcgacc tgcaggagct
     6421 cggcaaatac gagcagggat ctggatacat ccccgaggcc cccagagatg gccaggccta
     6481 cgtgcggaag gacggcgagt gggtactgct gagcacattc ctgggcagat ccggaggagg
     6541 cctggtgcca cagcagagcg gaggcctgaa cgacatcttc gaggcccaga agatcgagtg
     6601 gcacgagggc tgatgaggat ccagatctgc tgtgccttct agttgccagc catctgttgt
     6661 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6721 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6781 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6841 ggtgggctct atgggtaccc aggtgctgaa gaattgaccc ggttcctcct gggccagaaa
     6901 gaagcaggca catccccttc tctgtgacac accctgtcca cgcccctggt tcttagttcc
     6961 agccccactc ataggacact catagctcag gagggctccg ccttcaatcc cacccgctaa
     7021 agtacttgga gcggtctctc cctccctcat cagcccacca aaccaaacct agcctccaag
     7081 agtgggaaga aattaaagca agataggcta ttaagtgcag agggagagaa aatgcctcca
     7141 acatgtgagg aagtaatgag agaaatcata gaattttaag gccatgattt aaggccatca
     7201 tggccttaat cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7801 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8221 ctcggggggg gggggcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
     8281 gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat gagagctttg
     8341 ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg
     8401 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc
     8461 cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     8521 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     8581 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     8641 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     8701 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     8761 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
     8821 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     8881 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     8941 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     9001 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     9061 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     9121 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     9181 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     9241 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     9301 gcctcgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     9361 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     9421 agagattttg agacacaacg tggctttccc ccccccccca ttattgaagc atttatcagg
     9481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9541 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9601 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
