LOCUS       sequence-385179-        9636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the BJ.1 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 9636)
  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 9636)
  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-BJ.1-AVI
FEATURES             Location/Qualifiers
     source          1..9636
                     /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             6557..6601
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    6624..6848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7215..7232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7386..7974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7466..7485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8516..9331)
                     /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(8516..9331)
                     /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     9239..9258
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9323..9342
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     9609..9627
                     /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 tgatcacccg
     2881 aacccagtcc tacaccaaca gctttacacg gggcgtgtac taccctgaca aggtgttcag
     2941 atctagcgtc ctgcacagca ctcaggacct cttcctgccg ttcttcagca acgtgacatg
     3001 gttccacgcc atccacgtga gcggcacaaa cggaaccaag cggtttgata accccgccct
     3061 gccattcaat gatggagttt acttcgccag taccgagaag agtaacatca tccggggctg
     3121 gatcttcggc accaccctgg atagcaaaac acagagcctc ctgatcgtga acaatgccac
     3181 gaacgtcgtg atcaaggtgt gcgagttcca gttttgcaat gatcctttcc tggacgtgta
     3241 ccagaagaac aacaagagct ggatggaaag cgagttcaga gtctacagca gcgccaacaa
     3301 ctgcacattc gagtacgtct ctcagccttt tctgatggac cttgagggga aagaaggcaa
     3361 cttcaagaac ctgagagaat tcgtgttcaa gaacatcgac ggctacttca aaatctactc
     3421 caagcacaca cccatcaacc tggagcggga cctccctcag ggcttcagcg ccctggaacc
     3481 cctggtcgac ctgcccatag gcatcaacat aacgcggttc caaaccctgc tggccctgca
     3541 tagatcctac ctgactcctg gcgacagcag cagcggatgg accgccggag ctgcagccta
     3601 ctatgtgggc tacctgcaac ctagaacctt cctgctgaag tacaacgaga acggcacaat
     3661 cacagacgcc gtcgactgcg ccctggaccc tctctctgag acaaagtgca ccctgaagtc
     3721 cttcaccgtg gaaaagggca tctaccagac cagcaacttc cgggtgcagc ctacagagag
     3781 catcgtgcga tttccaaaca ttaccaacct ctgccccttc cacgaggtgt ttaacgccac
     3841 aacatttgcc tccgtttacg cctggaatag aaagagaatc agcaattgtg tggccgacta
     3901 ctccgtgatc tataacttcg cccctttctt cgccttcaag tgctacggcg tttccccaac
     3961 aaagctgaat gacctgtgct tcaccaacgt gtacgccgac tccttcgtaa ttagaggcaa
     4021 tgaggtgtcc cagatcgcac caggccagac cggtaacatc gctgactaca actataagct
     4081 gcctgatgat tttacaggct gcgtgatcgc ctggaactcc aacaagctgg atagcaagcc
     4141 tagcggcaac tacaactacc tgtaccggct gtttcgcaag tctaacctga aacctttcga
     4201 gagagacatc tccacagaga tctaccaggc cggtaacaag ccttgtaacg gggccgccgg
     4261 cttcaactgt tacgtccctc tgagaagcta cggcttccgg cctacctatg gagtcggcca
     4321 ccagccatac cgggtggtcg tgctgtcctt cgagttactc cacgcccccg ccaccgtctg
     4381 cggtcctaag aagtccacca atctggttaa gaacaaatgc gtgaacttca acttcaacgg
     4441 cctgaccggg accggcgtgc tgaccgaaag caacaaaaag ttcctcccct tccagcagtt
     4501 cggccgtgat atcgctgaca ccacagatgc cgtcagagat ccacagaccc tggaaatcct
     4561 ggatattaca ccctgctcct tcggaggagt ttctgtgatc acccccggga ccaataccag
     4621 caaccaggtg gctgtgctgt accaaggtgt taactgcacc gaggttcctg tggccatcca
     4681 cgccgatcag ctgacaccta cttggagagt gtactccact ggctccaatg tgttccagac
     4741 cagggccgga tgtctgatcg gcgccgagta cgtgaataac agttacgagt gcgacatccc
     4801 tatcggcgcc ggcatctgtg ccagctacca gacccagaca aagagccatg gcagcgcctc
     4861 ctctgtagct agccagagca tcatcgccta caccatgagc ctgggcgcag agaacagcgt
     4921 ggcctattcc aacaactcta tcgccattcc caccaacttt acaattagcg tcacaacaga
     4981 gatcctgccc gtgagcatga ccaagaccag cgtggactgt acaatgtaca tctgtggcga
     5041 cagcactgaa tgcagcaacc tgctgctgca atacggctcc ttttgcaccc aactgaagcg
     5101 ggcgctgacc ggaatcgccg tggaacagga caaaaatacc caggaggtgt tcgcccaagt
     5161 gaagcagatc tacaagaccc cacctatcaa gtacttcgac ggctttaact ttagccagat
     5221 tctccctgat ccttctaagc ctagcaagcg gagctttatc gaggatctgc tgttcaacaa
     5281 ggtcaccctg gccgatgccg gctttatcaa acagtatggc gattgtctgg gcgacatagc
     5341 cgccagagat ctgatctgcg cccagaaatt caacggcctg acagttctcc cacctctgct
     5401 gaccgacgag atgatcgctc agtacacctc tgccctgctg gctggcacca tcacatctgg
     5461 gtggacattt ggcgccggcg ccgccctgca gatccccttt gccatgcaga tggcctatag
     5521 attcaacgga atcggcgtga cccagaacgt gctgtatgaa aaccagaagc tgatcgctaa
     5581 ccagttcaat tctgccatcg gcaagatcca ggactccctc tcctctaccg ccagcgccct
     5641 gggcaaactg caggacgtgg tgaatcacaa cgcccaagcc ctgaacaccc tggtgaagca
     5701 gctcagcagc aagtttggcg ccatcagctc tgtgctgaac gatatcctgt ctagactgga
     5761 cccccctgaa gccgaagtcc agatcgatag actgatcaca ggcagactgc agatcctgca
     5821 aacctacgtg acccaacagc tgatcagggc cgctgaaata agagccagcg ccaatctcgc
     5881 cgctaccaag atgtccgagt gtgtgctggg acagtctaaa cgcgttgact tctgcggcaa
     5941 aggctatcac ctgatgagct tcccccagag cgcgccgcac ggcgtggtgt tcctgcatgt
     6001 gacatacgtg cctgcccaag agaagaattt cacaaccgcc cctgccatct gccacgacgg
     6061 caaggcccac ttcccaagag agggcgtttt cgtttccaat ggcacacact ggttcgtgac
     6121 acaaagaaac ttctacgaac cccagattat caccaccgac aacaccttcg tgagtggcaa
     6181 ttgtgacgtg gtcatcggaa tcgtgaacaa cacagtgtac gaccctctgc aacctgagct
     6241 ggactctttt aaggaagagc tggacaagta ctttaaaaac cacaccagtc ccgatgtgga
     6301 cctgggcgac atcagtggca ttaacgccag cgtggtgaac atccaaaagg aaatcgacag
     6361 actgaacgag gtggccaaga acctgaacga gtccctgatc gacctgcagg agctcggcaa
     6421 atacgagcag ggatctggat acatccccga ggcccccaga gatggccagg cctacgtgcg
     6481 gaaggacggc gagtgggtac tgctgagcac attcctgggc agatccggag gaggcctggt
     6541 gccacagcag agcggaggcc tgaacgacat cttcgaggcc cagaagatcg agtggcacga
     6601 gggctgatga ggatccagat ctgctgtgcc ttctagttgc cagccatctg ttgtttgccc
     6661 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6721 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     6781 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     6841 ctctatgggt acccaggtgc tgaagaattg acccggttcc tcctgggcca gaaagaagca
     6901 ggcacatccc cttctctgtg acacaccctg tccacgcccc tggttcttag ttccagcccc
     6961 actcatagga cactcatagc tcaggagggc tccgccttca atcccacccg ctaaagtact
     7021 tggagcggtc tctccctccc tcatcagccc accaaaccaa acctagcctc caagagtggg
     7081 aagaaattaa agcaagatag gctattaagt gcagagggag agaaaatgcc tccaacatgt
     7141 gaggaagtaa tgagagaaat catagaattt taaggccatg atttaaggcc atcatggcct
     7201 taatcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     8041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcggg
     8221 gggggggggc gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga
     8281 atcgccccat catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag
     8341 gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
     8401 agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt
     8461 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     8521 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     8581 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     8641 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     8701 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     8761 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     8821 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     8881 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     8941 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     9001 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     9061 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     9121 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     9181 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     9241 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     9301 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     9361 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     9421 tttgagacac aacgtggctt tccccccccc cccattattg aagcatttat cagggttatt
     9481 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9541 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     9601 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
