LOCUS       sequence-358143-        6558 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses SARS-CoV-2 RBD (aa324-526) domain 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 6558)
  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 6558)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-RBD(aa330-526)-B.1.1.529-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        276..655
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        656..859
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     810..830
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     856..880
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2789..2812
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3434..3478
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3501..3725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4092..4109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4263..4851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4343..4362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5393..6208)
                     /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(5393..6208)
                     /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     6116..6135
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6200..6219
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6486..6504
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6542..6558,1..6))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
       61 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      121 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      181 accgcatcag attggctatt ggccattgca tacgttgtat ccatatcata atatgtacat
      241 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
      301 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      361 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      421 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      481 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      541 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      601 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      661 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      721 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      781 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      841 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
      901 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccatcggc tcgcatctct
      961 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     1021 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     1081 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     1141 tccacgcttt gcctgaccct gcttgctcaa ctctagttaa cggtggaggg cagtgtagtc
     1201 tgagcagtac tcgttgctgc cgcgcgcgcc accagacata atagctgaca gactaacaga
     1261 ctgttccttt ccatgggtct tttctgcagt caccgtcgtc gacacgtgtg atcagatatc
     1321 gcggccgctc tagaccacca tgggatggtc atgtatcatc ctttttctag tagcaactgc
     1381 aaccggtgta cattccgcac ctgagctgct gggcggacct agtgtctttc tgttcccccc
     1441 caagcctaag gacaccctga tgatcagccg gacccccgag gttacatgcg tggtggtcga
     1501 tgtgtcccac gaggaccctg aagtgaagtt taattggtat gtggacggcg tcgaggtcca
     1561 caacgccaag accaagccta gagaggaaca gtacaacagc acctacagag tggtgagcgt
     1621 gctgaccgtg ctgcaccaag attggctgaa tggcaaggag tacaagtgca aggtgagcaa
     1681 caaggccctg ccagctccta ttgagaaaac catcagcaaa gccaagggcc agcctcggga
     1741 gccccaggtg tataccctgc ctccatctcg cgacgagctg accaagaacc aggtgtctct
     1801 gtactgcctg gtcaagggct tttaccctag cgacatcgcc gtggaatggg aaagcaacgg
     1861 ccagcctgag aacaactata aaacaacgcc acctgtgctg gactctgatg gcagcttctt
     1921 cctgtacagc aagctgaccg tggacaagag caggtggcaa cagggcaacg tgttcagctg
     1981 tagcgtgatg cacgaagctc tgcacaacca ctacacccag aaatctctca gcctgagtcc
     2041 cggcaagggc ggcggcggca gcggcggggg cggcagcggc ggcggtggat ctggcggagg
     2101 cggctccgcc cctgagcttc tgggcggccc ttctgtgttc ctgttccctc caaagcccaa
     2161 ggacacactg atgatcagca gaacccctga agtgacctgt gtggtggtgg acgtgtctca
     2221 tgaagatcct gaagtcaaat tcaactggta cgtggacggg gtggaagtgc acaatgccaa
     2281 aactaagcct cgggaagagc agtacaatag cacataccgg gtggtgtctg tactgaccgt
     2341 gctgcatcag gactggctga acggcaagga atacaagtgc aaggtgagca acaaggcact
     2401 gccagctccc atcgaaaaga ccatctccaa ggctaagggc cagcctagag agcctcaggt
     2461 gtacaccctg cctcctagca gagatgaact gacaaaaaac caggtgagcc tgacatgtct
     2521 ggttaagggc ttttacccct ctgacattgc cgtggaatgg gagtctaatg gacaacccga
     2581 aaacaactac aagaccacac cacctgtgct ggatagcgac ggcagcttct tcctgaccag
     2641 caagctgacc gtggataagt ccagatggca gcagggcaat gtgttcagct gcagcgtgat
     2701 gcacgaggct ctgcacaacc actacaccca gaaaagcctg tccctgagtc caggcaaggg
     2761 cggctctggc ggaggaggat ctggcggcct ggaagtcctc ttccagggcc ctcccaatat
     2821 cacaaacctg tgtcctttcg acgaggtgtt taacgccaca agattcgcca gcgtttatgc
     2881 ctggaataga aaaagaatct ctaactgcgt ggctgactac agcgtgctgt ataacctggc
     2941 cccttttttc accttcaagt gctacggcgt ttcccctaca aaactgaacg acctgtgctt
     3001 cacgaacgtg tatgccgaca gcttcgtgat ccggggagat gaggtgcggc agatcgcccc
     3061 tggccaaaca ggcaacatcg ccgattacaa ctacaagctg cccgacgact tcacaggatg
     3121 tgtgatcgcc tggaacagca acaagctcga cagcaaagtg agcggaaact acaactacct
     3181 gtacagactg tttagaaaga gcaatctgaa gcccttcgag agagatatca gcaccgagat
     3241 ctaccaggcc ggcaacaaac cgtgcaacgg cgtggccggc ttcaactgct acttccccct
     3301 gagaagctac tctttccggc ctacctacgg cgtgggccac cagccttaca gagtggtggt
     3361 gctgagcttc gagctgctgc acgcccctgc caccgtgtgc ggaggcggcg gcctggtgcc
     3421 ccagcagagc ggcggactga acgacatctt cgaggcccag aagatcgagt ggcacgaggg
     3481 ctgatgagga tccagatctg ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3541 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3601 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3661 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3721 tatgggtacc caggtgctga agaattgacc cggttcctcc tgggccagaa agaagcaggc
     3781 acatcccctt ctctgtgaca caccctgtcc acgcccctgg ttcttagttc cagccccact
     3841 cataggacac tcatagctca ggagggctcc gccttcaatc ccacccgcta aagtacttgg
     3901 agcggtctct ccctccctca tcagcccacc aaaccaaacc tagcctccaa gagtgggaag
     3961 aaattaaagc aagataggct attaagtgca gagggagaga aaatgcctcc aacatgtgag
     4021 gaagtaatga gagaaatcat agaattttaa ggccatgatt taaggccatc atggccttaa
     4081 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4141 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4201 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4261 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4381 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4441 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4501 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4561 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4621 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4681 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4741 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4801 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4861 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4921 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4981 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5041 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actcnggggg
     5101 ggggggcgct gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc
     5161 gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg
     5221 gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga
     5281 tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccgccgtccc
     5341 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     5401 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     5461 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     5521 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     5581 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     5641 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     5701 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     5761 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     5821 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     5881 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     5941 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     6001 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     6061 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     6121 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
     6181 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     6241 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     6301 gagacacaac gtggctttnc cccccccccc attattgaag catttatcag ggttattgtc
     6361 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6421 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6481 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6541 acctctgaca catgcagc
//
