LOCUS       sequence-354495-        9639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the B.1.1.529 (Omicron) 
            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.1.1.529-AVI
FEATURES             Location/Qualifiers
     source          1..9639
                     /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             6560..6604
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6627..6851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7218..7235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7389..7977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7469..7488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8519..9334)
                     /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(8519..9334)
                     /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     9242..9261
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9326..9345
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9612..9630
                     /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 tgcgtcaacc taacaacccg
     2881 gacccagctg cctcctgcct acacaaactc attcaccaga ggggtatact accctgacaa
     2941 ggtgttcaga agctccgtgc ttcacagcac ccaggatctg ttcctgcctt tcttctctaa
     3001 cgtgacctgg tttcacgtga tcagcggcac caacggcact aagcggtttg ataaccccgt
     3061 gctccctttc aacgacggcg tttacttcgc ctctatcgag aagagcaata tcatcagagg
     3121 ctggatcttc ggcaccaccc tggactccaa gacccagagc ctgctgatcg tgaacaacgc
     3181 caccaacgtg gtcatcaagg tgtgtgaatt tcagttctgc aacgaccctt ttctggacca
     3241 caagaataac aagagctgga tggaatctga gttccgggtg tacagctccg ccaacaattg
     3301 caccttcgag tacgtgtctc aacctttcct gatggacctg gaaggcaagc agggcaattt
     3361 caagaacctg cgggagttcg tgtttaaaaa catcgacggc tactttaaga tctatagcaa
     3421 gcacacaccc atcatcgtgc gggaacctga agatctgccc cagggcttct ctgccctgga
     3481 acccctggtg gacctgccta tcggcatcaa catcaccagg ttccaaaccc tgctggccct
     3541 gcatagatct tacctgacgc ccggcgatag tagctctggc tggaccgccg gagccgccgc
     3601 ttattacgtg ggctacctgc agcctagaac attcctcctg aagtacaacg agaacggtac
     3661 aatcacagat gccgttgatt gcgccctgga ccctctgagt gagactaaat gtaccctgaa
     3721 gagctttacc gtggaaaaag gcatttacca gacttccaac ttccgggtgc agcctaccga
     3781 gagcatcgtg agattcccta acatcacaaa cctgtgcccc ttcgacgagg tgttcaatgc
     3841 caccagattc gccagcgtgt atgcctggaa cagaaagcgc atcagcaatt gcgtggccga
     3901 ttacagcgtg ctgtacaacc tggctccttt ttttaccttc aagtgctacg gcgtctcccc
     3961 tacgaagctg aatgacctgt gcttcacaaa cgtgtacgcc gatagctttg tgatccgggg
     4021 cgacgaggtg agacagatcg cccctggcca gaccggaaac atagccgact acaactacaa
     4081 gctgccagac gacttcaccg gctgcgtgat agcctggaac agcaacaagc tggactccaa
     4141 ggtgtctggc aattacaact acctgtacag actgttcaga aagtccaacc tgaagccctt
     4201 tgagcgggat atcagcacag agatctacca ggccggaaac aagccctgta atggtgtggc
     4261 cggcttcaac tgctacttcc ccctgagatc ctacagcttc agacctacat atggcgtggg
     4321 ccaccagcct tacagagtgg tggtgctgtc tttcgagcta ctgcacgccc cagctacagt
     4381 gtgcggtcct aagaaaagca caaacctggt gaagaacaag tgcgtgaact tcaatttcaa
     4441 cggactgaag ggaacaggag tcctgaccga gtctaacaaa aagttcctgc cattccaaca
     4501 gttcggcaga gacatcgctg acaccacaga cgccgtgcgc gaccctcaga cgcttgagat
     4561 cctggacatc accccttgta gcttcggcgg agtcagcgtg atcacccccg gcaccaacac
     4621 atctaaccag gtggcagtgc tttatcaagg agtgaattgc accgaggtgc ctgtggctat
     4681 ccacgccgat cagctgaccc ctacctggcg ggtgtattct acaggctcga acgtgttcca
     4741 gaccagggct ggctgtctga tcggcgccga atacgtgaac aacagctacg agtgcgacat
     4801 ccccatcggc gccggcatct gcgccagcta tcagacacag actaaaagcc acggcagcgc
     4861 cagtagcgtt gccagccaga gcatcatcgc ctacaccatg agcctgggcg ctgaaaacag
     4921 cgtggcctac agcaacaaca gcatcgccat acccaccaac ttcaccatca gcgtgaccac
     4981 agagatcctg cctgtgtcca tgaccaagac cagcgtggac tgcacaatgt acatctgcgg
     5041 cgacagcacc gagtgcagca acctgctgct gcagtacggc tctttctgca cgcagctgaa
     5101 aagagccctg acaggcatcg ccgtggaaca ggataagaat acccaggagg tattcgccca
     5161 agtgaaacag atctacaaga ccccccccat caaatacttc ggaggcttca acttcagcca
     5221 gatcctgcca gaccccagca agccttctaa acggagcttc atcgaggacc tgctcttcaa
     5281 caaggtgacc ctggccgacg ccggcttcat caagcagtac ggagactgcc tgggcgacat
     5341 cgccgccaga gatctgattt gtgctcaaaa gttcaagggc ctgaccgtcc tgcctccact
     5401 gctgacagac gaaatgatcg ctcagtacac cagcgccctg ctggccggca ccatcacctc
     5461 cggctggaca tttggagccg gcgccgcttt acagatcccc tttgccatgc agatggccta
     5521 ccggttcaac ggcatcggcg tgacccagaa cgtcctgtac gaaaaccaga aactgattgc
     5581 caaccagttt aacagcgcca tcggaaagat ccaggacagc ctcagcagca ccgcctccgc
     5641 cttgggaaaa ctgcaggacg tggtaaatca caacgcccag gccctgaaca ccctggtgaa
     5701 gcagctgagc agcaagtttg gtgccatctc cagcgtgctt aacgacatct tcagcagatt
     5761 ggaccctcct gaagccgaag tccagatcga tagactgatc acaggcagac tgcagtccct
     5821 gcaaacctac gtgacccaac agctgatcag ggccgctgaa ataagagcca gcgccaatct
     5881 cgccgctacc aagatgtccg agtgtgtgct gggacagtct aaacgcgttg acttctgcgg
     5941 caaaggctat cacctgatga gcttccccca gagcgcgccg cacggcgtgg tgttcctgca
     6001 tgtgacatac gtgcctgccc aagagaagaa tttcacaacc gcccctgcca tctgccacga
     6061 cggcaaggcc cacttcccaa gagagggcgt tttcgtttcc aatggcacac actggttcgt
     6121 gacacaaaga aacttctacg aaccccagat tatcaccacc gacaacacct tcgtgagtgg
     6181 caattgtgac gtggtcatcg gaatcgtgaa caacacagtg tacgaccctc tgcaacctga
     6241 gctggactct tttaaggaag agctggacaa gtactttaaa aaccacacca gccccgatgt
     6301 ggacctgggc gacatcagtg gcattaacgc cagcgtggtg aacatccaaa aggaaatcga
     6361 cagactgaac gaggtggcca agaacctgaa cgagtccctg atcgacctgc aggagctcgg
     6421 caaatacgag cagggatctg gatacatccc cgaggccccc agagatggcc aggcctacgt
     6481 gcggaaggac ggcgagtggg tactgctgag cacattcctg ggcagatccg gaggaggcct
     6541 ggtgccacag cagagcggag gcctgaacga catcttcgag gcccagaaga tcgagtggca
     6601 cgagggctga tgaggatcca gatctgctgt gccttctagt tgccagccat ctgttgtttg
     6661 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     6721 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     6781 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     6841 gggctctatg ggtacccagg tgctgaagaa ttgacccggt tcctcctggg ccagaaagaa
     6901 gcaggcacat ccccttctct gtgacacacc ctgtccacgc ccctggttct tagttccagc
     6961 cccactcata ggacactcat agctcaggag ggctccgcct tcaatcccac ccgctaaagt
     7021 acttggagcg gtctctccct ccctcatcag cccaccaaac caaacctagc ctccaagagt
     7081 gggaagaaat taaagcaaga taggctatta agtgcagagg gagagaaaat gcctccaaca
     7141 tgtgaggaag taatgagaga aatcatagaa ttttaaggcc atgatttaag gccatcatgg
     7201 ccttaatctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     8101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     8161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8221 gggggggggg ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc
     8281 tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg
     8341 taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg
     8401 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
     8461 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     8521 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     8581 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     8641 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     8701 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     8761 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     8821 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     8881 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     8941 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     9001 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     9061 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     9121 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     9181 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     9241 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     9301 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     9361 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     9421 gattttgaga cacaacgtgg ctttcccccc ccccccatta ttgaagcatt tatcagggtt
     9481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9541 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9601 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
