LOCUS       sequence-314420-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD domain 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   Zhou T, Teng IT, Olia AS, Cerutti G, Gorman J, Nazzari A, Shi W, 
            Tsybovsky Y, Wang L, Wang S, Zhang B, Zhang Y, Katsamba PS, Petrova 
            Y, Banach BB, Fahad AS, Liu L, Lopez Acevedo SN, Madan B, Oliveira 
            de Souza M, Pan X, Wang P, Wolfe JR, Yin M, Ho DD, Phung E, DiPiazza
            A, Chang LA, Abiona OM, Corbett KS, DeKosky BJ, Graham BS, Mascola 
            JR, Misasi J, Ruckwardt T, Sullivan NJ, Shapiro L, Kwong PD
  TITLE     Structure-Based Design with Tag-Based Purification and In-Process 
            Biotinylation Enable Streamlined Development of SARS-CoV-2 Spike 
            Molecular Probes.
  JOURNAL   Cell Rep. 2020 Oct 27;33(4):108322. doi: 
            10.1016/j.celrep.2020.108322. Epub 2020 Oct 12.
  PUBMED    33091382
REFERENCE   2  (bases 1 to 6558)
  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: "Cell Rep.";
            date: "2020-10-27"; volume: "33(4):108322. doi"; pages: " 
            10.1016/j.celrep.2020.108322. Epub 2020 Oct 12"
COMMENT     Sequence Label: pVRC8400-SARS-CoV-2-RBD-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        490..693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     644..664
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     690..714
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2623..2646
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3268..3312
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3335..3559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3926..3943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4097..4685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4177..4196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5227..6042)
                     /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(5227..6042)
                     /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     5950..5969
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6034..6053
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6320..6338
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6376..6398)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6498..6517
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtaaggaga aaataccgca tcagattggc tattggccat tgcatacgtt gtatccatat
       61 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      661 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      721 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccat
      781 cggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
      841 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
      901 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
      961 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctag ttaacggtgg
     1021 agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga cataatagct
     1081 gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt cgtcgacacg
     1141 tgtgatcaga tatcgcggcc gctctagacc accatgggat ggtcatgtat catccttttt
     1201 ctagtagcaa ctgcaaccgg tgtacattcc gcaccagagc tgctgggagg acctagcgtg
     1261 ttcctgtttc cccctaagcc taaggacacc ctgatgatca gccggacccc agaggtgaca
     1321 tgcgtggtgg tggacgtgtc ccacgaggac cccgaggtga agttcaactg gtacgtggat
     1381 ggcgtggagg tgcacaatgc caagaccaag ccaagggagg agcagtacaa ctctacctat
     1441 cgcgtggtga gcgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtacaag
     1501 tgtaaggtgt ctaataaggc cctgcctgcc ccaatcgaga agaccatcag caaggcaaag
     1561 ggacagccaa gggagcctca ggtgtatacc ctgccaccca gccgcgacga gctgacaaag
     1621 aaccaggtgt ccctgtactg cctggtgaag ggcttctatc cttccgatat cgccgtggag
     1681 tgggagtcta atggccagcc agagaacaat tacaagacca cacctccagt gctggactcc
     1741 gatggctctt tctttctgta ttccaagctg accgtggata agagccggtg gcagcagggc
     1801 aacgtgttta gctgttccgt gatgcacgag gccctgcaca atcactacac acagaagtct
     1861 ctgagcctgt cccccggcaa gggaggagga ggcagcggcg gaggaggctc cggaggcggc
     1921 ggctctggcg gcggcggcag cgcccctgaa ctgctgggag gaccaagcgt gttcctgttt
     1981 ccacctaagc ccaaagatac cctgatgatc tccaggaccc ctgaagtcac ctgcgtggtc
     2041 gtggacgtgt ctcatgagga ccccgaagtc aagttcaact ggtacgtgga cggagtcgaa
     2101 gtccataacg ccaagaccaa gcctagagaa gagcagtaca actccaccta tagagtggtg
     2161 tctgtgctga cagtgctgca tcaggattgg ctgaatggca aggaatataa gtgtaaggtg
     2221 agcaataagg ccctgcccgc ccctatcgag aagaccatct ctaaagccaa aggacagcca
     2281 agggagccac aggtgtatac actgccaccc agccgggacg agctgaccaa gaaccaggtg
     2341 agcctgacat gtctggtcaa gggcttctac cccagtgaca ttgctgtgga gtgggagtcc
     2401 aatggccagc ctgagaacaa ttataaaact accccaccag tgctggacag cgatggctcc
     2461 ttctttctga ccagcaagct gacagtggat aagtcccgct ggcagcaggg aaacgtgttt
     2521 tcttgtagcg tgatgcatga ggccctgcat aaccactaca cccagaagtc cctgtctctg
     2581 agcccaggca agggaggcag cggaggcgga ggcagcggag gcctggaggt gctgtttcag
     2641 ggccctccca atatcacaaa cctgtgccct tttggcgagg tgttcaacgc aaccaggttc
     2701 gccagcgtgt acgcatggaa taggaagcgc atctccaact gcgtggccga ctattctgtg
     2761 ctgtacaaca gcgcctcctt ctctaccttt aagtgctatg gcgtgagccc cacaaagctg
     2821 aatgacctgt gctttaccaa cgtgtacgcc gattccttcg tgatcagggg cgacgaggtg
     2881 cgccagatcg caccaggaca gacaggcaag atcgcagact acaattataa gctgcctgac
     2941 gatttcaccg gctgcgtgat cgcctggaac tctaacaatc tggatagcaa agtgggcggc
     3001 aactacaatt atctgtaccg gctgtttaga aagtctaatc tgaagccatt cgagagggac
     3061 atctccacag aaatctacca ggccggctct accccctgca atggcgtgga gggctttaac
     3121 tgttatttcc ctctgcagag ctacggcttc cagccaacaa acggcgtggg ctatcagccc
     3181 taccgcgtgg tggtgctgtc ttttgagctg ctgcacgcac ctgcaacagt gtgcggagga
     3241 ggaggcctgg tgccacagca gagcggaggc ctgaacgaca tcttcgaggc ccagaagatc
     3301 gagtggcacg agggctgatg aggatccaga tctgctgtgc cttctagttg ccagccatct
     3361 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3421 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3481 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3541 gatgcggtgg gctctatggg tacccaggtg ctgaagaatt gacccggttc ctcctgggcc
     3601 agaaagaagc aggcacatcc ccttctctgt gacacaccct gtccacgccc ctggttctta
     3661 gttccagccc cactcatagg acactcatag ctcaggaggg ctccgccttc aatcccaccc
     3721 gctaaagtac ttggagcggt ctctccctcc ctcatcagcc caccaaacca aacctagcct
     3781 ccaagagtgg gaagaaatta aagcaagata ggctattaag tgcagaggga gagaaaatgc
     3841 ctccaacatg tgaggaagta atgagagaaa tcatagaatt ttaaggccat gatttaaggc
     3901 catcatggcc ttaatcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3961 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4021 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4081 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4141 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4201 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4261 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4321 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4381 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4441 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4501 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4561 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4621 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4681 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4741 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4801 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4861 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4921 cctgactcgg gggggggggg cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat
     4981 accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg ttgatgagag
     5041 ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg gaacggtctg
     5101 cgttgtcggg aagatgcgtg atctgatcct tcaactcagc aaaagttcga tttattcaac
     5161 aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa
     5221 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     5281 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     5341 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     5401 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     5461 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     5521 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     5581 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     5641 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     5701 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     5761 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     5821 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     5881 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     5941 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     6001 tcgcggcctc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     6061 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     6121 acatcagaga ttttgagaca caacgtggct ttcccccccc ccccattatt gaagcattta
     6181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6301 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6361 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6421 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6481 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6541 tgaaataccg cacagatg
//
