LOCUS       sequence-314424-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD-L455RG496R 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-L455RG496R-AVI
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     122..141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        292..671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        672..875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     826..846
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..896
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2805..2828
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3450..3494
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    3517..3741
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4108..4125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4359..4378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5409..6224)
                     /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(5409..6224)
                     /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     6132..6151
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6216..6235
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6502..6520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6558,1..22))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
       61 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
      121 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
      181 tgcgtaagga gaaaataccg catcagattg gctattggcc attgcatacg ttgtatccat
      241 atcataatat gtacatttat attggctcat gtccaacatt accgccatgt tgacattgat
      301 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      361 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      421 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      481 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      541 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      601 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      661 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      721 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      781 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      841 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      901 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
      961 atcggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     1021 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     1081 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     1141 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct agttaacggt
     1201 ggagggcagt gtagtctgag cagtactcgt tgctgccgcg cgcgccacca gacataatag
     1261 ctgacagact aacagactgt tcctttccat gggtcttttc tgcagtcacc gtcgtcgaca
     1321 cgtgtgatca gatatcgcgg ccgctctaga ccaccatggg atggtcatgt atcatccttt
     1381 ttctagtagc aactgcaacc ggtgtacatt ccgcaccaga gctgctggga ggacctagcg
     1441 tgttcctgtt tccccctaag cctaaggaca ccctgatgat cagccggacc ccagaggtga
     1501 catgcgtggt ggtggacgtg tcccacgagg accccgaggt gaagttcaac tggtacgtgg
     1561 atggcgtgga ggtgcacaat gccaagacca agccaaggga ggagcagtac aactctacct
     1621 atcgcgtggt gagcgtgctg acagtgctgc accaggactg gctgaacggc aaggagtaca
     1681 agtgtaaggt gtctaataag gccctgcctg ccccaatcga gaagaccatc agcaaggcaa
     1741 agggacagcc aagggagcct caggtgtata ccctgccacc cagccgcgac gagctgacaa
     1801 agaaccaggt gtccctgtac tgcctggtga agggcttcta tccttccgat atcgccgtgg
     1861 agtgggagtc taatggccag ccagagaaca attacaagac cacacctcca gtgctggact
     1921 ccgatggctc tttctttctg tattccaagc tgaccgtgga taagagccgg tggcagcagg
     1981 gcaacgtgtt tagctgttcc gtgatgcacg aggccctgca caatcactac acacagaagt
     2041 ctctgagcct gtcccccggc aagggaggag gaggcagcgg cggaggaggc tccggaggcg
     2101 gcggctctgg cggcggcggc agcgcccctg aactgctggg aggaccaagc gtgttcctgt
     2161 ttccacctaa gcccaaagat accctgatga tctccaggac ccctgaagtc acctgcgtgg
     2221 tcgtggacgt gtctcatgag gaccccgaag tcaagttcaa ctggtacgtg gacggagtcg
     2281 aagtccataa cgccaagacc aagcctagag aagagcagta caactccacc tatagagtgg
     2341 tgtctgtgct gacagtgctg catcaggatt ggctgaatgg caaggaatat aagtgtaagg
     2401 tgagcaataa ggccctgccc gcccctatcg agaagaccat ctctaaagcc aaaggacagc
     2461 caagggagcc acaggtgtat acactgccac ccagccggga cgagctgacc aagaaccagg
     2521 tgagcctgac atgtctggtc aagggcttct accccagtga cattgctgtg gagtgggagt
     2581 ccaatggcca gcctgagaac aattataaaa ctaccccacc agtgctggac agcgatggct
     2641 ccttctttct gaccagcaag ctgacagtgg ataagtcccg ctggcagcag ggaaacgtgt
     2701 tttcttgtag cgtgatgcat gaggccctgc ataaccacta cacccagaag tccctgtctc
     2761 tgagcccagg caagggaggc agcggaggcg gaggcagcgg aggcctggag gtgctgtttc
     2821 agggccctcc caatatcaca aacctgtgcc cttttggcga ggtgttcaac gcaaccaggt
     2881 tcgccagcgt gtacgcatgg aataggaagc gcatctccaa ctgcgtggcc gactattctg
     2941 tgctgtacaa cagcgcctcc ttctctacct ttaagtgcta tggcgtgagc cccacaaagc
     3001 tgaatgacct gtgctttacc aacgtgtacg ccgattcctt cgtgatcagg ggcgacgagg
     3061 tgcgccagat cgcaccagga cagacaggca agatcgcaga ctacaattat aagctgcctg
     3121 acgatttcac cggctgcgtg atcgcctgga actctaacaa tctggatagc aaagtgggcg
     3181 gcaactacaa ttatctgtac cggaggttta gaaagtctaa tctgaagcca ttcgagaggg
     3241 acatctccac agaaatctac caggccggct ctaccccctg caatggcgtg gagggcttta
     3301 actgttattt ccctctgcag agctaccgct tccagccaac aaacggcgtg ggctatcagc
     3361 cctaccgcgt ggtggtgctg tcttttgagc tgctgcacgc acctgcaaca gtgtgcggag
     3421 gaggaggcct ggtgccacag cagagcggag gcctgaacga catcttcgag gcccagaaga
     3481 tcgagtggca cgagggctga tgaggatcca gatctgctgt gccttctagt tgccagccat
     3541 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3601 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3661 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3721 gggatgcggt gggctctatg ggtacccagg tgctgaagaa ttgacccggt tcctcctggg
     3781 ccagaaagaa gcaggcacat ccccttctct gtgacacacc ctgtccacgc ccctggttct
     3841 tagttccagc cccactcata ggacactcat agctcaggag ggctccgcct tcaatcccac
     3901 ccgctaaagt acttggagcg gtctctccct ccctcatcag cccaccaaac caaacctagc
     3961 ctccaagagt gggaagaaat taaagcaaga taggctatta agtgcagagg gagagaaaat
     4021 gcctccaaca tgtgaggaag taatgagaga aatcatagaa ttttaaggcc atgatttaag
     4081 gccatcatgg ccttaatctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4681 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5101 tgcctgactc gggggggggg ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc
     5161 ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag
     5221 agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc
     5281 tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca
     5341 acaaagccgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc
     5401 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5461 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5521 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5581 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5641 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     5701 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5761 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     5821 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5881 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     5941 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     6001 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     6061 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     6121 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     6181 aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     6241 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     6301 taacatcaga gattttgaga cacaacgtgg ctttcccccc ccccccatta ttgaagcatt
     6361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6481 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6541 ggtgatgacg gtgaaaac
//
