LOCUS       sequence-314422-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-L455RA475R 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-L455RA475R-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        203..582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        583..786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     737..757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     783..807
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2716..2739
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3361..3405
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3428..3652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4019..4036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5320..6135)
                     /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(5320..6135)
                     /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     6043..6062
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6127..6146
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6413..6431
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6469..6491)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
       61 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcagatt
      121 ggctattggc cattgcatac gttgtatcca tatcataata tgtacattta tattggctca
      181 tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
      241 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      421 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      481 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      541 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      601 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      661 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      721 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      841 catagaagac accgggaccg atccagcctc catcggctcg catctctcct tcacgcgccc
      901 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
      961 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     1021 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     1081 tgaccctgct tgctcaactc tagttaacgg tggagggcag tgtagtctga gcagtactcg
     1141 ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg ttcctttcca
     1201 tgggtctttt ctgcagtcac cgtcgtcgac acgtgtgatc agatatcgcg gccgctctag
     1261 accaccatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
     1321 tccgcaccag agctgctggg aggacctagc gtgttcctgt ttccccctaa gcctaaggac
     1381 accctgatga tcagccggac cccagaggtg acatgcgtgg tggtggacgt gtcccacgag
     1441 gaccccgagg tgaagttcaa ctggtacgtg gatggcgtgg aggtgcacaa tgccaagacc
     1501 aagccaaggg aggagcagta caactctacc tatcgcgtgg tgagcgtgct gacagtgctg
     1561 caccaggact ggctgaacgg caaggagtac aagtgtaagg tgtctaataa ggccctgcct
     1621 gccccaatcg agaagaccat cagcaaggca aagggacagc caagggagcc tcaggtgtat
     1681 accctgccac ccagccgcga cgagctgaca aagaaccagg tgtccctgta ctgcctggtg
     1741 aagggcttct atccttccga tatcgccgtg gagtgggagt ctaatggcca gccagagaac
     1801 aattacaaga ccacacctcc agtgctggac tccgatggct ctttctttct gtattccaag
     1861 ctgaccgtgg ataagagccg gtggcagcag ggcaacgtgt ttagctgttc cgtgatgcac
     1921 gaggccctgc acaatcacta cacacagaag tctctgagcc tgtcccccgg caagggagga
     1981 ggaggcagcg gcggaggagg ctccggaggc ggcggctctg gcggcggcgg cagcgcccct
     2041 gaactgctgg gaggaccaag cgtgttcctg tttccaccta agcccaaaga taccctgatg
     2101 atctccagga cccctgaagt cacctgcgtg gtcgtggacg tgtctcatga ggaccccgaa
     2161 gtcaagttca actggtacgt ggacggagtc gaagtccata acgccaagac caagcctaga
     2221 gaagagcagt acaactccac ctatagagtg gtgtctgtgc tgacagtgct gcatcaggat
     2281 tggctgaatg gcaaggaata taagtgtaag gtgagcaata aggccctgcc cgcccctatc
     2341 gagaagacca tctctaaagc caaaggacag ccaagggagc cacaggtgta tacactgcca
     2401 cccagccggg acgagctgac caagaaccag gtgagcctga catgtctggt caagggcttc
     2461 taccccagtg acattgctgt ggagtgggag tccaatggcc agcctgagaa caattataaa
     2521 actaccccac cagtgctgga cagcgatggc tccttctttc tgaccagcaa gctgacagtg
     2581 gataagtccc gctggcagca gggaaacgtg ttttcttgta gcgtgatgca tgaggccctg
     2641 cataaccact acacccagaa gtccctgtct ctgagcccag gcaagggagg cagcggaggc
     2701 ggaggcagcg gaggcctgga ggtgctgttt cagggccctc ccaatatcac aaacctgtgc
     2761 ccttttggcg aggtgttcaa cgcaaccagg ttcgccagcg tgtacgcatg gaataggaag
     2821 cgcatctcca actgcgtggc cgactattct gtgctgtaca acagcgcctc cttctctacc
     2881 tttaagtgct atggcgtgag ccccacaaag ctgaatgacc tgtgctttac caacgtgtac
     2941 gccgattcct tcgtgatcag gggcgacgag gtgcgccaga tcgcaccagg acagacaggc
     3001 aagatcgcag actacaatta taagctgcct gacgatttca ccggctgcgt gatcgcctgg
     3061 aactctaaca atctggatag caaagtgggc ggcaactaca attatctgta ccggaggttt
     3121 agaaagtcta atctgaagcc attcgagagg gacatctcca cagaaatcta ccagcgcggc
     3181 tctaccccct gcaatggcgt ggagggcttt aactgttatt tccctctgca gagctacggc
     3241 ttccagccaa caaacggcgt gggctatcag ccctaccgcg tggtggtgct gtcttttgag
     3301 ctgctgcacg cacctgcaac agtgtgcgga ggaggaggcc tggtgccaca gcagagcgga
     3361 ggcctgaacg acatcttcga ggcccagaag atcgagtggc acgagggctg atgaggatcc
     3421 agatctgctg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3481 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3541 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3601 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat gggtacccag
     3661 gtgctgaaga attgacccgg ttcctcctgg gccagaaaga agcaggcaca tccccttctc
     3721 tgtgacacac cctgtccacg cccctggttc ttagttccag ccccactcat aggacactca
     3781 tagctcagga gggctccgcc ttcaatccca cccgctaaag tacttggagc ggtctctccc
     3841 tccctcatca gcccaccaaa ccaaacctag cctccaagag tgggaagaaa ttaaagcaag
     3901 ataggctatt aagtgcagag ggagagaaaa tgcctccaac atgtgaggaa gtaatgagag
     3961 aaatcataga attttaaggc catgatttaa ggccatcatg gccttaatct tccgcttcct
     4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4981 cagcgatctg tctatttcgt tcatccatag ttgcctgact cggggggggg gggcgctgag
     5041 gtctgcctcg tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca
     5101 gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga
     5161 ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat
     5221 ccttcaactc agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt
     5281 aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat
     5341 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     5401 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     5461 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     5521 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     5581 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     5641 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     5701 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     5761 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     5821 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     5881 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     5941 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     6001 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     6061 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg
     6121 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     6181 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     6241 gctttccccc cccccccatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6421 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6481 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6541 tcagggcgcg tcagcggg
//
