LOCUS       sequence-369456-        9639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 S2P protein from the BA.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 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-BA.2-AVI
FEATURES             Location/Qualifiers
     source          1..9639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(124..939)
                     /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(124..939)
                     /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     847..866
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     931..950
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1217..1235
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1273..1295)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1395..1414
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1565..1944
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1945..2148
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2099..2119
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2145..2169
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4078..4101
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             7804..7848
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    7871..8095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8462..8479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8633..9221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8713..8732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa
       61 gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
      121 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
      181 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      241 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      301 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      361 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
      421 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      481 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
      541 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
      601 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
      661 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
      721 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
      781 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
      841 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
      901 cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
      961 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     1021 tcagagattt tgagacacaa cgtggctttn cccccccccc cattattgaa gcatttatca
     1081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1141 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     1201 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     1261 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1321 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1381 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     1441 aataccgcac agatgcgtaa ggagaaaata ccgcatcaga ttggctattg gccattgcat
     1501 acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac attaccgcca
     1561 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     1621 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1681 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     1741 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     1801 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     1861 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1921 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     1981 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2041 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2101 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2161 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     2221 cgatccagcc tccatcggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
     2281 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
     2341 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
     2401 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
     2461 tctagttaac ggtggagggc agtgtagtct gagcagtact cgttgctgcc gcgcgcgcca
     2521 ccagacataa tagctgacag actaacagac tgttcctttc catgggtctt ttctgcagtc
     2581 accgtcgtcg acacgtgtga tcagatatcg cggccgctct agaccaccat gggatggtca
     2641 tgtatcatcc tttttctagt agcaactgca accggtgtac attccgcacc agagctgctg
     2701 ggaggaccta gcgtgttcct gtttccccct aagcctaagg acaccctgat gatcagccgg
     2761 accccagagg tgacatgcgt ggtggtggac gtgtcccacg aggaccccga ggtgaagttc
     2821 aactggtacg tggatggcgt ggaggtgcac aatgccaaga ccaagccaag ggaggagcag
     2881 tacaactcta cctatcgcgt ggtgagcgtg ctgacagtgc tgcaccagga ctggctgaac
     2941 ggcaaggagt acaagtgtaa ggtgtctaat aaggccctgc ctgccccaat cgagaagacc
     3001 atcagcaagg caaagggaca gccaagggag cctcaggtgt ataccctgcc acccagccgc
     3061 gacgagctga caaagaacca ggtgtccctg tactgcctgg tgaagggctt ctatccttcc
     3121 gatatcgccg tggagtggga gtctaatggc cagccagaga acaattacaa gaccacacct
     3181 ccagtgctgg actccgatgg ctctttcttt ctgtattcca agctgaccgt ggataagagc
     3241 cggtggcagc agggcaacgt gtttagctgt tccgtgatgc acgaggccct gcacaatcac
     3301 tacacacaga agtctctgag cctgtccccc ggcaagggag gaggaggcag cggcggagga
     3361 ggctccggag gcggcggctc tggcggcggc ggcagcgccc ctgaactgct gggaggacca
     3421 agcgtgttcc tgtttccacc taagcccaaa gataccctga tgatctccag gacccctgaa
     3481 gtcacctgcg tggtcgtgga cgtgtctcat gaggaccccg aagtcaagtt caactggtac
     3541 gtggacggag tcgaagtcca taacgccaag accaagccta gagaagagca gtacaactcc
     3601 acctatagag tggtgtctgt gctgacagtg ctgcatcagg attggctgaa tggcaaggaa
     3661 tataagtgta aggtgagcaa taaggccctg cccgccccta tcgagaagac catctctaaa
     3721 gccaaaggac agccaaggga gccacaggtg tatacactgc cacccagccg ggacgagctg
     3781 accaagaacc aggtgagcct gacatgtctg gtcaagggct tctaccccag tgacattgct
     3841 gtggagtggg agtccaatgg ccagcctgag aacaattata aaactacccc accagtgctg
     3901 gacagcgatg gctccttctt tctgaccagc aagctgacag tggataagtc ccgctggcag
     3961 cagggaaacg tgttttcttg tagcgtgatg catgaggccc tgcataacca ctacacccag
     4021 aagtccctgt ctctgagccc aggcaaggga ggcagcggag gcggaggcag cggaggcctt
     4081 gaggtactgt ttcagggccc tcaatgtgtg aacctgatca ccaggaccca gagctataca
     4141 aactccttta caagaggcgt gtactatcct gacaaagtgt tcagaagcag cgtgctgcac
     4201 tccacccagg acctgttcct gccttttttc agcaacgtga cgtggttcca cgccatccac
     4261 gtgtctggca ccaacggcac caagcggttc gacaaccccg tgctgccttt caacgacggc
     4321 gtgtacttcg ccagcaccga gaagagcaat atcatccgcg gctggatctt cggcacaacc
     4381 ctggactcta agacccagag cctgctgatc gtgaacaatg ccacaaacgt agtgatcaag
     4441 gtgtgtgaat tccagttctg taacgaccct ttcctggatg tctactatca caagaacaac
     4501 aagtcctgga tggaaagcga gttccgggtc tactcaagcg ccaacaactg cacgttcgag
     4561 tacgtgagcc agcctttcct gatggacctg gagggcaagc aaggcaactt caagaacctg
     4621 cgggaattcg tgttcaagaa catcgatgga tacttcaaga tctacagcaa gcacactcct
     4681 atcaatctgg gcagagatct gcctcagggc ttcagcgccc tggaacctct ggtcgatctg
     4741 cctatcggca tcaacatcac cagattccag acactgctgg ccctgcacag aagctatctc
     4801 acaccaggcg attcctctag cggatggaca gccggggccg ccgcttacta cgtgggctac
     4861 ctgcagccta gaacattcct gctaaagtac aacgagaacg gcaccatcac cgacgcagtg
     4921 gactgcgcgc tggacccgct aagcgagaca aaatgcacac tgaaatcctt taccgtggaa
     4981 aagggcatct accagacatc caacttccgg gtccagccca ccgagtccat agtgcggttc
     5041 cctaacatta ccaacctgtg ccccttcgac gaggtgttca acgccaccag attcgccagc
     5101 gtgtacgcct ggaatagaaa acggatcagc aactgcgtgg ccgattacag cgtgctctac
     5161 aacttcgccc ctttcttcgc tttcaagtgt tacggcgtga gccccacaaa gctgaacgac
     5221 ctgtgcttca caaacgtgta cgccgattct ttcgtgatca gaggcaacga ggtgtcccag
     5281 atcgcccccg gccagaccgg aaatatcgct gactacaact acaagttacc agacgacttc
     5341 accggatgtg tgatcgcctg gaacagcaac aagctggaca gcaaggttgg cggaaactac
     5401 aattacctgt atagactgtt ccggaagagc aacctgaagc cctttgagag agatatttct
     5461 acagagatct accaggccgg aaacaagccc tgcaacggag tggcggggtt caactgctac
     5521 ttccctctga gaagctacgg cttcagacct acgtacggcg tgggccacca gccttacaga
     5581 gtggtggtgc tgagctttga actgctgcac gcccctgcca ctgtgtgcgg tcctaagaaa
     5641 agcacaaacc ttgtgaagaa caagtgcgtg aactttaact tcaacggact gactggcacc
     5701 ggcgtgctga ccgagagcaa caaaaagttc ctgccttttc agcagttcgg cagagatatc
     5761 gccgacacca cagacgccgt gcgggacccc cagaccctgg aaatcctgga catcacacct
     5821 tgtagcttcg gaggcgtgag cgtaatcacc cctggtacca acacctccaa ccaggtggcc
     5881 gttctgtacc agggcgtgaa ttgcaccgaa gtgcccgtgg ctatacacgc cgatcagctg
     5941 accccaacat ggagagtgta cagcacagga agcaacgtgt tccaaactag agccggctgc
     6001 ctgatcggcg ccgagtacgt gaacaacagc tacgagtgcg acatacctat cggcgctgga
     6061 atttgtgcct cttatcagac acaaaccaag agccacggca gcgcttcatc cgttgcttct
     6121 cagagcatca tcgcgtatac aatgagcctg ggcgccgaaa actcggtggc ctacagcaac
     6181 aatagcatcg ccatcccaac caatttcacc atttctgtga ctaccgaaat cctgcctgtg
     6241 agcatgacca aaacctccgt ggattgcacc atgtacatct gtggcgactc tacagaatgc
     6301 agcaacctcc tgctccagta cggttctttc tgcacccagc tgaaaagagc cctgacaggc
     6361 atcgctgtgg aacaggacaa gaatacccag gaggtcttcg cccaggtcaa acaaatctac
     6421 aagacacctc caatcaagta cttcggcggc ttcaacttta gccagatcct gcccgaccca
     6481 tctaagccca gcaagcgttc ttttatcgag gacctgctgt ttaataaggt gaccctggcc
     6541 gacgccggct tcatcaagca gtacggcgac tgcctgggcg atattgctgc tagagacctg
     6601 atctgcgccc agaagttcaa tggcctgacc gtactgcctc ctctgctgac cgatgagatg
     6661 atcgcccaat acaccagcgc cctcctggcc ggcacaatca ccagtggctg gaccttcgga
     6721 gctggcgccg ctctgcagat cccattcgcc atgcagatgg cctaccggtt caatggcatc
     6781 ggagttaccc agaacgtgct gtacgagaac caaaagctga tcgccaatca gttcaactct
     6841 gcaataggca agatccagga ctccctgagc agcacagcct cagccctggg gaagctgcaa
     6901 gatgtggtca accacaacgc ccaggccctg aacaccctgg tgaaacagct gagcagcaag
     6961 tttggagcca tcagcagcgt gctgaacgac atcctgtctc gactggatcc tcctgaagcc
     7021 gaagtgcaga tcgatcgtct tatcaccggc aggctgcagt ccctgcagac ctacgtgacc
     7081 cagcagctga tcagggccgc cgagatcaga gcctctgcca acttggccgc caccaagatg
     7141 agtgaatgcg tgctgggcca gagcaagcgg gtggacttct gcggcaaagg ctaccacctg
     7201 atgtccttcc cccagtctgc cccacacggc gtggttttcc tccatgtgac atatgtgcct
     7261 gctcaggaga agaacttcac caccgcccct gctatctgcc acgacggcaa ggcccacttc
     7321 ccaagagagg gcgtgttcgt gagtaatggc acacactggt tcgtgaccca gcggaacttc
     7381 tacgagcctc agatcatcac cacagataac acctttgtgt ccggcaattg tgacgtggtc
     7441 atcggaatcg tgaacaatac cgtgtacgac cctctgcagc ccgaactgga tagcttcaaa
     7501 gaggaactgg acaagtactt taaaaaccat accagccccg atgtggacct gggagatatc
     7561 agcggcatca acgccagcgt tgtcaacatt cagaaagaaa tcgacagact caacgaggtg
     7621 gccaagaacc tgaacgagtc cctgatcgac ctgcaggagc tgggaaagta cgagcagggc
     7681 tcaggctaca tccccgaggc ccccagagat ggccaggcct acgtgcggaa ggacggcgag
     7741 tgggtactgc tgagcacatt cctgggcaga tccggaggag gcctggtgcc acagcagagc
     7801 ggaggcctga acgacatctt cgaggcccag aagatcgagt ggcacgaggg ctgatgagga
     7861 tccagatctg ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     7921 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     7981 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     8041 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgggtacc
     8101 caggtgctga agaattgacc cggttcctcc tgggccagaa agaagcaggc acatcccctt
     8161 ctctgtgaca caccctgtcc acgcccctgg ttcttagttc cagccccact cataggacac
     8221 tcatagctca ggagggctcc gccttcaatc ccacccgcta aagtacttgg agcggtctct
     8281 ccctccctca tcagcccacc aaaccaaacc tagcctccaa gagtgggaag aaattaaagc
     8341 aagataggct attaagtgca gagggagaga aaatgcctcc aacatgtgag gaagtaatga
     8401 gagaaatcat agaattttaa ggccatgatt taaggccatc atggccttaa tcttccgctt
     8461 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8521 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8581 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8641 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8701 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8761 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8821 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8881 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8941 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9001 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9061 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9121 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actcgggggg gggggncgct
     9481 gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat
     9541 ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg
     9601 tgattttgaa cttttgcttt gccacggaac ggtctgcgt
//
